Evidence-Based Complementary and Alternative Medicine

Evidence-Based Complementary and Alternative Medicine / 2016 / Article
Special Issue

New Exploration of Chinese Herbal Medicines in Hepatology

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Review Article | Open Access

Volume 2016 |Article ID 4750163 | https://doi.org/10.1155/2016/4750163

Hor-Yue Tan, Serban San-Marina, Ning Wang, Ming Hong, Sha Li, Lei Li, Fan Cheung, Xiao-Yan Wen, Yibin Feng, "Preclinical Models for Investigation of Herbal Medicines in Liver Diseases: Update and Perspective", Evidence-Based Complementary and Alternative Medicine, vol. 2016, Article ID 4750163, 26 pages, 2016. https://doi.org/10.1155/2016/4750163

Preclinical Models for Investigation of Herbal Medicines in Liver Diseases: Update and Perspective

Academic Editor: Yoshiji Ohta
Received22 Oct 2015
Revised23 Dec 2015
Accepted30 Dec 2015
Published28 Jan 2016

Abstract

Liver disease results from a dynamic pathological process associated with cellular and genetic alterations, which may progress stepwise to liver dysfunction. Commonly, liver disease begins with hepatocyte injury, followed by persistent episodes of cellular regeneration, inflammation, and hepatocyte death that may ultimately lead to nonreversible liver failure. For centuries, herbal remedies have been used for a variety of liver diseases and recent studies have identified the active compounds that may interact with liver disease-associated targets. Further study on the herbal remedies may lead to the formulation of next generation medicines with hepatoprotective, antifibrotic, and anticancer properties. Still, the pharmacological actions of vast majority of herbal remedies remain unknown; thus, extensive preclinical studies are important. In this review, we summarize progress made over the last five years of the most commonly used preclinical models of liver diseases that are used to screen for curative herbal medicines for nonalcoholic fatty liver disease, liver fibrosis/cirrhosis, and liver. We also summarize the proposed mechanisms associated with the observed liver-protective, antifibrotic, and anticancer actions of several promising herbal medicines and discuss the challenges faced in this research field.

1. Introduction

Hepatic disease refers to a constellation of disorders of the liver that can lead to decompensated liver function. The liver is a very important organ that is mainly responsible for vital functions such as detoxification and glucose and lipid metabolism as well as the synthesis of many key enzymes that regulate these metabolic processes. Acute liver disease is defined as a rapid hepatic dysfunction that occurs in the absence of previous history of chronic liver disease; it is caused, for example, by excessive consumption of antibiotics or acetaminophen. By contrast, chronic liver disease is a long-term dynamic process that involves persistent hepatocytic destruction and regeneration. Major risk factors for chronic liver disease are hepatitis B viral and hepatitis C viral (HBV and HCV) infection and alcoholic liver-induced injury leading to alcoholic liver disease (ALD) as well as a constellation of metabolic disorders that can lead to nonalcoholic fatty liver disease (NAFLD). Liver exposure to these risk factors gradually results in hepatocytic injury associated with tissue infiltration of inflammatory cells and altered transcriptome in the affected cell populations. As a result, both liver scarring and regeneration are triggered which, if left unchecked, will ultimately progress to profound changes in liver architecture and liver cirrhosis. In addition, patients with cirrhosis have a higher risk of developing hepatocellular carcinoma (HCC) [1].

The incidence of NAFLD is highest among all chronic liver diseases in the United States where it was responsible for 75% of all cases in 2008 [2]. Globally, the prevalence of NAFLD ranges from 10 to 35% depending on different diagnostic tools and populations studied. For nonalcoholic induced steatohepatitis (NASH), between 3 and 5% of the global population is at risk [3]. In clinic, pioglitazone or vitamin E is only given to patients with advanced stage of NASH who failed lifestyle intervention due to the potential risk of the treatment in inducing stroke [4]. Global mortality from liver cirrhosis rose to over 1 million in 2010, accounting for 2% of all deaths worldwide. Approximately 16 out of every 100,000 people died due to liver cirrhosis worldwide, and the incidence is greater in South and Central Asia as well as Eastern European countries [5]. Liver transplantation remains the only intervention for patients with liver cirrhosis, as alternative drug therapies are not available in the clinic. Antifibrotic therapy is emerging as a possible option as several antifibrotic candidates have been shortlisted preclinically and await further study [6]. Similar to liver cirrhosis, surgical removal and liver transplantation are the most effective treatment for HCC. However, not all patients are suitable for liver surgery as the cancer may have spread and the 5-year survival rate was reported to be about 15% for patients diagnosed with HCC [7]. There is currently an unfilled medical need to find alternatives to liver transplantation by combating inflammation and the production of reactive oxygen species that are key aspects of chronic liver diseases. In many countries like China, there is a rich history of using herbal medicine to treat liver diseases. Due to the antioxidant and anti-inflammatory nature of these botanicals, their active ingredients could lead to the development of novel hepatoprotective, antifibrotic, and antiliver cancer therapies. To date, plant-based products such as Fuzheng Huayu formula and silymarin have been well documented for use in liver diseases [8]. Fuzheng Huayu formula, an FDA-approved Chinese medicinal formulation, is now undergoing phase IV clinical trial for patients with cirrhosis secondary to HBV infection in China and phase II clinical trial had been completed for chronic hepatitis C patient in US [9]. Although some herbal remedies hold promise for attenuating or reversing the progression of liver diseases, other compounds may be toxic and damage the liver [10]. Therefore, preclinical studies of dose escalation and efficacy testing should be thoroughly conducted in animal models in order to provide a better understanding of safety and efficacy before advancing herbal remedies to clinical studies.

Appropriate in vitro models for NAFLD and liver fibrosis remain to be developed. Furthermore, despite extensive use of immortalized cell lines and primary cultures, effective systems for high throughput drug screening are lacking. In part, this is due to the multifactorial nature of liver disease that is not amenable to investigation through simple in vitro cell models.

In this review, we aim to systematically review the most commonly used animal models that were employed to screen and study the efficacy of herbal medicines for liver diseases. This may serve as a guiding tool for selecting the appropriate liver disease model for herbal medicine screening as well as facilitating further exploration of studied herbal remedies for clinical applications.

2. Animal Models for NAFLD and NASH

NAFLD is an idiopathic pathological condition where excessive fat deposition in the liver is caused by factors other than chronic alcohol consumption. If untreated, NAFLD may further progress to nonalcoholic steatohepatitis or NASH and then liver fibrosis/cirrhosis and eventually liver cancer. NAFLD is more prevalent in patients with obesity, diabetes, insulin resistance, and hypertension, which suggests that it is the manifestation of an untreated underlying metabolic dysfunction.

Steatosis or the abnormal accumulation of fats in hepatocytes is thought to be the original disease-causing event leading to further dysfunction through oxidative stress, fatty acid and inflammatory cytokine-mediated liver injury, and apoptosis as well as changed lipid partitioning [11]. Steatosis divides into two general types: macrovesicular and microvesicular, which differ with respect to the number and size of lipid vacuoles and to the position of the nucleus in the cytoplasm. In macrovesicular steatosis, lipid vacuoles are large and the nucleus tends to push aside, while in microvesicular steatosis the nucleus is usually not affected. NAFLD accounts for approximately 5% of all liver steatosis [12]. However, in NASH, several other pathologies were observed, such as hepatocellular ballooning and intralobular and inflammatory infiltration with immune cells [13].

As mentioned earlier, liver pathology can progress from obesity and insulin resistance to macrovesicular/microvesicular steatosis, hepatocellular ballooning, and intralobular inflammation. At the biochemical level, there is a malfunction in lipid metabolism consisting of increased long chain fatty acid influx and altered fatty acid synthesis as well as decreased triglyceride production. Broadly, two types of animal models for NAFLD/NASH have been described but neither faithfully reflects the human conditions. In one type of the model, the disease is induced by a genetic modification that targets any one of the processes mentioned above while in the other it is caused by altered dietary intake. The ob/ob, db/db, adiponectin null, and KK-Ay models have been routinely employed to study the development of NAFLD. However, these models do not directly progress to steatohepatitis but require additional insults, such as various dietary treatments or the injection of hepatotoxins. On the other hand, diet-based models such as the methionine and choline deficiency (MCD) model or the high fat diet (HFD) model are easier to develop and present some of the histological features of human diseases. However, disease development in these models can vary substantially based on the composition of the diets and the duration of experiments.

The HFD model is most commonly used to explore the effect of herbal medicines on NAFLD and NASH. After 4 months, total fat/body weight was found to be five times higher in experimental HFD mice than in naïve mice. HFD mice had higher total serum cholesterol, triglyceride, and insulin levels as well as impaired glucose tolerance as compared to naïve mice. All these suggest pathological initiation of NAFLD. Administration of rhein, from Rheum palmatum L. after 40 days of HFD, normalized liver fat levels and improved insulin resistance [14]. In another study by Xiao et al. [15], the histopathological modifications of NASH such as hepatocytic necrosis and infiltration with inflammatory cells were observed after 8 weeks of HFD. Collagen formation detected by Sirius red staining also suggested that the disease was progressing from steatohepatitis to hepatic fibrosis. Treatment with Lycium barbarum polysaccharides for 4 weeks reduced insulin resistance and obesity as well as improving the histopathological changes incurred by HFD. Herbal medicines have also shown promising results in the MCD mouse model, where as previously mentioned, methionine and choline are absent from diet [16]. These components are required for beta-oxidation and VLDL synthesis and their dietary suppression leads to fatty acid accumulation and oxidative stress in hepatocytes. A recent study showed that, after eight weeks of MCD, there was observable macrovesicular steatosis, inflammation, and hepatocytic necrosis. Administration of Fuzheng Huayu formula ameliorated these changes through downregulation of CYP2E1 and HO-1, markers of oxidative stress. In addition, TNF-α and IL-6 were similarly downregulated in this study [17].

Animal models in which liver disease is induced by a combination of selected genetic backgrounds, diets, and the administration of hepatotoxins have also been proposed [18, 19]. In the study by Ma et al., ApoE () mice with HFD showing signs of NAFLD had ameliorated pathological changes after being fed with Huanglian Jiedu extract [20]. One possible explanation for this effect is promotion of phagocytosis by the increased population of M2 macrophages. Similarly, administration of the Japanese herbal medicines Sho-saiko-to and Juzen-taiho-to in MCD-fed db/db mice for 4 weeks reduced lobular inflammation and liver ballooning [21]. In the above models, SREBP-1c, adiponectin, PGC-1 alpha, and FABP were frequently used as biochemical markers to monitor lipid and energy metabolism functions. Overexpression of SREBP-1c induced lipid synthesis and reduced VLDL efflux and lipid oxidation while increasing triglyceride deposition in hepatocytes [22]. Adiponectin is another important cytokine that regulates glucose metabolism and fatty acid breakdown. It is reported that susceptibility for liver fibrosis in adiponectin knockout mice is higher compared to naïve mice [23]. A Japanese herbal medicine, bofutsushosan, ameliorated hepatic steatosis and inflammation through increased plasma adiponectin levels and reduced SREBP-1c expression. Simultaneously, bofutsushosan also enhanced fatty acid oxidation and reduced inflammation through upregulation of α/λ-PPAR [24]. Similar findings were reported for ping-tang recipe that greatly enhanced α/λ-PPAR expression while reducing that of the lipogenic genes, SREBP-1c, FAS, and L-FABP [25]. Proposed mechanisms for the observed effects of several herbal medicines against NAFLD and NASH together with the investigated animal models are summarized in Table 1.


Model (ingredients)DurationHerbal medicinePathological and biochemical changes; mechanisms involvedReference

Diet induced models (high calories/fats diet)

10% lard oil and 2% cholesterol4 wkSi Jun Zi Tang (SJZ), Lizhong Tang (LZ), Linggui Zhugan Tang (LGZG), and Shen Zhuo Tang (SZ)Reduced epididymal fat index and hepatic fats infiltration; reduced triglycerides and ALT levels.[84]

Methionine and choline bitartrate tablets4 wkAlkaloids of Rubus alceifoliusReduced hepatic lobule, serum ALT, AST, TNFα, and IL-6; hepatic SOD and MDA.[16]

25% lard, 2% cholesterol 0.5% sodium cholate, and 25% Tween-8056 dSchisandra chinensis BaillReduced SOD, serum TC, and LDLC and increased hepatic MDA. [85]

12% lard oil, 2% cholesterol, 0.2% propylthiouracil, and 0.5% bile salt 6 wkSapindus mukorossi Gaertn.Increased fat deposition, fat-storing cells proliferation, collagen accumulation, macrovesicular steatosis, ballooning degeneration, and cytoplasmic vacuolation (model group). Reduced liver cell volume, hepatic lobules, and fat droplets (treatment group). Reduced TC, TG, LDL-C, ALT, and AST; increased APN level in treatment group. [86]

10% lard, 2% cholesterol, 0.2% bile salts, 15% sucrose, 8% baking soybean powder 7 wkTZQ formula (red peony root, mulberry leaf, lotus leaf, danshen root, and hawthorn leaf)Increased hepatic steatosis, necrosis, and inflammatory cell infiltration (model group). Reduced TC, TG, LDL-C, HDL-C, and TC-HDL/HDL in treatment group. [87]

88% normal chow plus 10% lard plus 2% cholesterol4 wk + 4 wk treatmentJiang Zhi GranuleImproved hepatic steatosis, reduced ALT and AST, and improved free fatty acid and triacylglycerol levels. Reduced LXR alpha and SREBP-1c. [88]

MCD diet 8 wkFuzheng Huayu recipeModel group showed disoriented lobule, macrosteatosis, hepatocyte necrosis, and inflammatory infiltration. Treatment group reduced ALT, AST and P450 2E1, TNFα, and IL-6. Increased HO-1. Inhibited alpha-SMA, TGF-β1, and Col I and Col III. [17]

High fat diet8 wkPing-tang recipeSuppressed visceral fat accumulation, enhanced glucose metabolism, and ameliorated hepatic steatosis. Upregulated PGC-1 alpha, PPAR alpha, and gamma and reduced SREBP-1c, FAS, and FABP; activated AMPK and acetyl CoA carboxylase phosphorylation.[25]

High fat and high fructose diet9 wkPuerariae radix, Lycium barbarum, Crataegus pinnatifida, and Polygonati rhizomaReduced fasting blood glucose and improved insulin resistance. [89]

10% lard, 1.5% cholesterol, 0.2% sodium deoxycholate, 5% sugar, 0.05% prothiopyrimidine10 wk (5 wk treatment)Yiqi huoxue decoction and solutions of herbsHepatic fat deposit, macrovesicular steatosis, ballooning degeneration, and cytoplasmic vacuolation (model group); reduced lipid degeneration, fat droplets and smaller liver cells, and delineated hepatic lobules (treatment group). [90]

9.3 g AIN-93MX, 2.6 g AIN-93VX, 0.5 g choline bitartrate, 1.1 g DL-methionine, 57.5 g lactalbumin hydrolysate, 117.5 g dextrose, 36.6 mL fish oil, and 4.5 g suspending agent K 12 wkLycium barbarum polysaccharidesReduced lipid droplet accumulation, inflammatory cells infiltration, and hepatocyte necrosis as observed in model group. Reduced phosphorylation of TGF-β1 and α-SMA. Decreased SREBP-1c and PPARγ2, but reduced ATGL and adiponectin; restored antioxidant enzymes; reduced NF-κB, p-p38 MAPK, and enhanced autophagy.[15]

High fat diet12 wkChinese medicine recipesTreatment group blocked fatty degeneration. Decreased expressions of JNK and p-JNK. [91]

5.3 kcal/g (fat 59%, protein 16%, and carbohydrate 24%)15 wkYin-Chen-Hao-TangReduced hepatocyte foaming and ballooning. Reduced TNF-α and MCP-1. Promoted senescence marker protein-30 metabolism. Restored oxidative stress markers.[92]

45 kcal% fat, 20 kcal% protein, and 35 kcal% carbohydrate16 wkGarcinia cambogia supplementIncreased collagen deposition in treatment group. Reduced obesity, but induced hepatic fibrosis (inflammation TNF-α and MCP-1 and oxidative stress SOD, GSH-Px, and TBARS increased).[93]

Axungia porci 10%, cholesterol 1.5%, and bile salt 0.5% 16 wkChaihu-Shugan-San and Shen-ling-bai-zhu-SanModel group showed microvesicular and macrovesicular steatosis, lobular, and portal inflammation and hepatocyte ballooning while treatment group improved them. Decreased TNF-alpha and IL-6 in serum, inhibited TLR4, and activation of p38 MAPK.[94]

High fat diet16 wkSoothing liver and invigorating spleen recipesReduced TC, LDL-C, TC, and TG in treatment group. Reduced TLR4 expression. [95]

21.9 kJ/g, 60% fat, 20% protein, and 20% carbohydrate20 wkTroxerutinDecreased epididymal adipose tissue mass, lipid accumulation, and lipid levels in treatment group. Suppressed NAD (+) depletion, increased NAMPT, and decreased PARP1. Increased SirT1 and AMPK and inhibited mTORC1 and Lpin 1β/α.[96]

60% of kcal as fat with an energy density of 5.24 kcal/g24 wkCrude extract of Lycium barbarum polysaccharideLowered TC, LDL, TG, and DAG levels. Induced phosphorylation of AMPK, reduced nuclear expression of SREBP-1c, and increased UCP1 and PGC-1 alpha in adipose tissue.[62]

60% kcal from lard/soybean 9.8 : 14 mo + 40 d treatmentRheinSteatotic and enlarged hepatocytes (model group); reduced lipid accumulation (treatment group). Reduced ALT, TC, LDL, and TG level in treatment group and improved insulin resistant. Suppressed SREBP-1c and LXR; inhibited T-BET, and enhanced GATA-3 and pSTAT3. [14]

15% fat, 15% sucrose, and 2% cholesterol12 wkSalvia-Nelumbinis naturalisReduced macrovesicular steatosis, TG, LDL-C, and FFA levels. Increased IRS and Akt phosphorylation and decreased SOCS3.[97]

High fat dietCigu Xiaozhi pillsModel group showed adipose degeneration and inflammatory cell infiltration. Treatment group reduced TG, TC, ALT, AST, and MDA level while increased SOD and GPX. Reduced TNFα.[98]

Models developed by combined insults

ApoE (−/−) + high cholesterol diet4 wkHuanglian Jiedu decoctionAmeliorated pathological changes in fatty liver. Associated with increase of M2 macrophages populations.[20]

MCD diet in db/db mice4 wkSho-saiko-to (TJ-9), inchin-ko-to (TJ-135), juzen-taiho-to (TJ-48), and keishi-bukuryo-gan (TJ-25)TJ-9 and TJ-48 reduced ALT. Necroinflammation, hepatic lobules, steatosis, and ballooning degeneration. Reduced TGF-β1, increased TNFα, IL-6, and PPAR-gamma, and reduced MDA.[21]

High fat diet  + streptozotocin i.p.4 wkpomegranate flowers polyphenolsReduced fat drops, non-HDL-C, and transaminase. Antioxidant ability enhanced and PON1 increased in liver.[19]

High fat diet (10% lard and 2% cholesterol) + CCl48 wkDangyao (Swertia pseudochinensis Hara) and Shuifeiji (Silybum marianum Gaertn.)Ameliorated hepatosteatosis lobules ballooning degeneration and inflammatory infiltration as observed in model group. Reduced ALT, AST, TAG, and MDA. Increased UCP2.[18]

High fat diet (88% normal chow + 10% lard + 2% cholesterol) + CCl48 wkDangfei Liganning capsulesImproved steatosis, hepatocyte ballooning, and inflammatory infiltration as observed in model group. Improved MDA and ALT. [99]

10% lard + 2% cholesterol + CCl48 wkSoothing liver and invigorating spleen recipesReduced SREBP-1 and SCD-1.[100]

High cholesterol diet in KK-Ay mice10 wkCorosolic acidReduced blood cholesterol and liver cholesterol content. Inhibited activity of cholesterol acyltransferase.[101]

High fat diet (640 kcal/100 g) + gold-thioglucose12 wkBofutsushosanBlocked hepatic steatosis, inflammation, hepatocyte ballooning, and Mallory-Denk bodies. Reduced ALT, AST, and TG levels. Induced adiponectin and its receptors, increased PPAR-α and PPAR-γ, and decreased SREBP-1c; reduced IR by phosphorylated Akt.[24]

HF diet (19.6% carbohydrates, 18.2% proteins, and 62.2% lipids; total energy, 506 kcal/100) and 30% sucrose in drinks 12 wkGoshajinkiganReduced AST. Increased body and adipose tissue weight and reduced elevated liver weight. [102]

Content not specified in the paper.

3. Animal Models for Liver Fibrosis and Cirrhosis

Liver fibrosis is a progressive disease that is caused by viral induced hepatitis, alcoholic induced liver injury, NAFLD/NASH, chronic biliary retention, or parasitic infection induced injury [26]. The underlying pathophysiological process consists of overlapping cycles of wound healing and cell necrosis that ultimately leads to accumulation of extracellular matrix containing collagen and other matrix components [27]. An important event in fibrogenesis is the formation of hepatic myofibroblasts that secrete collagen. If left unchecked, the process results in the complete destruction of liver architecture with hepatic insufficiency that is the marker of a more severe process of cirrhosis. Ultimately, portosystemic shunting leads to liver failure [28]. To date, there is no effective treatment for liver fibrosis or cirrhosis. Whether or not these processes are reversed or arrested in their progression is a matter of current controversy. Yet, animal studies have shown that in some cases experimentally induced liver fibrosis can be reversed using botanical extracts.

Liver fibrogenesis can commonly be induced in animal settings by cholestasis or administration of hepatotoxins. In some studies, fibrosis was induced using longer-term models of alcoholic or nonalcoholic liver injury or by a combination of the two [29]. Recently, models of gene-modified mice were established by knocking down the MDR2 gene or by overexpressing TGF-β1 [30, 31]. The administration of hepatotoxins to induce liver injury in rodents is a model that is commonly used in herbal medicine studies. Liver injury by carbon tetrachloride (CCl4) administration is more frequently used than thioacetamide (TAA) or dimethyl/diethyl nitrosamine (DEN) due to short onset of disease development and the direct cytotoxic effect of CCl4 on hepatocytes. CCl4 induced early fibrosis can be detected within two weeks after the first administration, and 5 to 7 weeks are sufficient to detect all the physiological symptoms of liver fibrosis. Continuous administration of CCl4 will eventually lead to liver cirrhosis. The immediate cytotoxic effect of CCl4 on hepatocytes recruits inflammatory cells and induces secretion of proinflammatory cytokines leading to necrosis. Scarring occurs as a result of persistent cycles of cell death and is followed by hepatocytic regeneration/proliferation processes [27]. A study by Zhou et al. [32] showed that 6 weeks of intraperitoneal CCl4 injection 3 times per week resulted in multiple scarring/fibrotic events such as bridged vessels, fibrous septa, and even regenerative nodules. Treatment with Xuefuzhuyu starting at week 4 ameliorated the hepatic stellate cells (HSC) activation and ECM formation and reduced expression of α-SMA and collagen I. Another study conducted by Shen et al. [33] also showed that treatment with Diwu Yanggan in a 6-week CCl4 model attenuated epithelial-to-mesenchymal cellular transition, a commonly observed condition in fibrogenesis, through upregulation of E-cadherin and downregulation of vimentin. Compared to CCl4, disease modelling using TAA and DEN (a carcinogen) requires longer exposure time to induce HCC. In all these cases and irrespective of the causative agent, disease development is remarkably similar across the models. For example, intraperitoneal injection of TAA for 6 weeks increased fibrous septa, portal tract, and liver sinusoids as well as α-SMA staining of liver cells, the same as in the CCl4 model. In the TAA model, administration of kaerophyllin from Bupleurum scorzonerifolium reversed histopathological changes and further reduced the levels of proinflammatory cytokines TNF-α, IL-1β, and MCP-1 [34].

It is worth mentioning that Fuzheng Huayu formula, a traditional Chinese medicine that showed antifibrotic effects in clinical practice, also demonstrated significant in vivo effect in DEN and CCl4 models [3538]. Currently, the Fuzheng Huayu formula is in a phase IV multicenter clinical trial for HBV induced liver cirrhosis [39]. In DMN-induced mice, administration of Fuzheng Huayu formula reduced inflammatory cell infiltration and collagen accumulation and lowered α-SMA expression. The antifibrotic effect of the extract is believed to downregulate TGF-β1 and p-Smad2/3 in injured tissue [35] as well as attenuating apoptosis via TNFα blockade [36]. Our research group has extensively studied the Coptidis rhizoma aqueous extract (CRAE), a traditional Chinese medicine used in clinical practice for liver disease. Simultaneous administration of CRAE and CCl4 for 8 weeks alleviated the formation of fibrous septa, pseudo lobes, and collagen deposition. The effect is comparable to that of bear bile, a medicine traditionally used for liver disease, which is also effective in CCl4 induced, cholestatic, and alcohol fed murine models [40]. The protective effect of Coptidis rhizoma extracts may be secondary to ameliorating oxidative stress and decreasing apoptosis [41]. Proposed mechanisms of action of herbal medicines in liver fibrosis/cirrhosis and the models used to test them are summarized in Table 2.


ModelExperiment durationsHerbal medicinePathological and biochemical changes; mechanism involvedReference

Hepatotoxins

Thioacetamide induced6 wksKaerophyllinReduced collagen accumulation and fibrosis score. Reduced ALT, AST, and α-SMA. Reduced TNFα, IL-1β, MCP-1, ICAM-1; increased PPAR-γ.[34]
7 wksSilybinReduced Tbil, ALT, and AST levels and necrotic zones and steatosis. Reversed loss of CYP3A and PXR; reduced α-SMA and improved anti-inflammatory cytokines secretion. [103]
8 wksEthanolic extract of rhizomes of Z. officinale  [104]
8/12 wksMagnesium lithospermate BReduced ALT, AST and α-SMA, TGF-β1, and collagen-α1 (I). Inhibited NF-κB activation, MCP-1, and H2O2 induced ROS production. [105]
12 wksChunggan extractReduced inflammation, collagen deposition, and large septa formation. Reduced ALP, AST and Bil, MDA, iNOS, and TNFα; increased TGF-β. [106]

DEN/DMN induced3 wksJia-wei-xiao-yao-sanReduced SOD, lipid peroxidation, and xanthine oxidase activity. [107]
4 wksYinchenhao decoction (YCHD group) and Yiguanjian (YGJ group)Reduced Hyp and α-SMA. Reduced TNFα, PDGF, MDA, and GST activity; increased L-FABP and transferrin. [108]
4 wksHuangqi decoctionReduced hepatocyte apoptosis. Decreased apoptotic effectors, cleaved-caspase-3, α-SMA, and TGF-β1. [109]
4 wksYinchenhao decoction, Yinchen Wuling San, and Zhizi Baipi decoctionReduced Hyp content and Tbil level. [110]
4 wksAnluohuaxianwanReduced ALT, AST, and contents of HA. Increased MMP-2.[111]
4 wksHuangqi decoction[112]
4 wksFuzheng Huayu and Huangqi decoction[38]
4 wksFuzheng Huayu recipeReduced Hyp content. Reduced TGF-β1, its receptor, and p-Smad2/3. [35]
4 wksXiaopi pillDecreased ALT, AST, ALP and Tbil, and α-SMA. Reduced TGF-β1, TIMP-1, and HO-1.[113]
4 wksModified SinisanReduced liver injury biomarkers.[114]
4 wksYi Guan JianReduced fibrotic septa, degenerated hepatocytes, and collagen deposition. Reduced collagen α1-I, TIMP-1, and α-SMA. [115]
6 wksVaccinium corymbosum L.Decreased liver apoptosis, necrosis, and proliferation. Reduced hepatic lipid peroxidation, protein oxidation, and nitrotyrosine levels; increased GST activity.[116]
6 wksBoswellia serrata and Salvia miltiorrhiza extractsShowed normal liver parenchymal architecture and only mild fibrosis.
Reduced α-SMA, collagen I– collagen III, CTGF, TGF-β1, Smad3, and Smad7.
[117]
15 wksGexiazhuyu decoctionDecreased GOT and GPT. Reduced collagen α-1 and α-SMA.[118]
Corbrin Shugan capsule[119]

CCl4
induced
4 wksBiejiayinzi, Gexiazhuyu Tang, and Fugan Wan Gexiazhuyu Tang and Fugan Wan treatment improved inflammatory necrosis and fat degeneration. Reduced collagen accumulation. [120]
6 wksFuzhenghuayu decoctionDecreased ALT, AST, and Tbil. Reduced area ratio of liver fibrosis and α-SMA. [37]
6 wksXiayuxue decoctionDecreased Sirius red positive area. Reduced α-SMA and type I collagen. [121]
6 wksDiwu YangganReduced ALT, AST, Hyp, and collagen deposition and tissue damage. Increased E-cadherin, TGF-nc; reduced vimentin, BMP-7, Hh ligand Shh, receptor Smo and Ptc, and Gli1. [33]
6 wksXuefuzhuyu decoctionReduced ALT, AST, Tbil, and Hyp. No ECM deposition and reduced necroinflammatory foci. Decreased α-SMA, collagen I, CD31, VEGF, VEGFR-2, HIF-1 alpha, and ADMA; increased DDAH1. [32]
7 wksEthanol extract of Cortex DictamniImproved pathological grading. Reduced collagen deposition and Hyp content. Increased pY-STAT1.[122]
8 wksBaicalinDecreased Hy, steatosis, liver necrosis, and fibrotic septa formation. Reduced TGF-β1 and PPARγ. [123]
8 wksCoptidis rhizoma aqueous extractReduced Tbil and AST. Improved histological changes. Reduced SOD and Erk1/2 inhibition[40]
8 wksCichorium intybus L. extractReduced ALT, AST, Hyp, and histopathological changes. Increased GSH, SOD; reduced MDA. Reduced TGF-β1 and α-SMA. [124]
8 wksFuzhenghuayu decoctionReduced ALT, AST, and hepatocyte apoptosis. Decreased collagen deposition and inflammatory cell infiltration. Reduced α-SMA and Hyp.[37]
8 wksAcremoniumterricola milleretal myceliumDecreased HA, laminin, and procollagen type III levels, Hyp. Improved pathological changes. Restored SOD and GSH-Px, inhibited lipid peroxidation. Decreased TGF-β, Smad2/3 phosphorylation and increased Smad7 inhibitor. [125]
8 wksRougan Huaqian granulesDecreased AST and HA. Reduced α-SMA, LN, Col I, Col III, Col IV, and MMP-2.[126]
8 wksFufang Biejia Ruangan pillsDecreased ALT, AST. Reduced collagen deposition and improved hepatic lesion. Reduced hyaluronic acid Col IV, type III procollagen laminin, TGF-β1, and Smad3.[127]
8 wksGanfukangDecreased ALT and AST. Ameriolated ductular proliferation. Reduced α-SMA, MMP-2 and TIMP-1, synthesis of collagen, and activation of the Wnt/beta-catenin.[128]
8 wksHuisheng oral solutionInhibited collagen formation and improved liver function. Reduced Smad3, TGF-β1, α-SMA, and TIMP-1. [129]
8 wksMethanol extracts of Ficus carica Linn. (Moraceae) leaves and fruits and Morus alba Linn. root barksReduced ALT, AST and ALP, and total bilirubin. Improved hepatocellular architecture. Restored antioxidant related content. [130]
9 wksDahuangzhechong pillDecreased ALT, AST, HA, laminin, type IV collagen, and procollagen III and reverses hepatic fibrosis. Reduced α-SMA, serum TNF ASIL-13, p38MAPK, and Erk phosphorylation. [131]
9 wksXiayuxue decoctionInhibited liver injury, fatty degeneration, and collagen deposition. Blocked CD31, vWF, VEGF, VEGFR2, DAF, α-SMA, and MMP-2 and MMP-9 activities. [132]
9 wksYiguanjian decoctionIncreased Cu/Zn SOD, DJ-1, glutathione S-transferase Yb-1 subunit, and aldo-keto reductase family 7, A2.[133]
9 wksYiguanjianIncreased SOD, Prxd6, transferrin, and L-FABP; decreased MDA, HSP70, and HO-1.[67]
9 wksYiguanjian decoctionReduced collagen deposition. Suppressed α-SMA, Col I, TIMP-1, TIMP-2, MMP-13, and MMP-14 and activities of MMP-2 and MMP-9.[68]
12 wksHuganjiexian decoctionInhibited collagen type I, collagen type III, TGF-β1, and PDGF-BB.[134]
12 wksXiaozheng Rongmu powderChanges in ALT, AST, and Tbil are milder and with hepatocytes mitosis. [135]
12 wksShuganjianpifangDecreased steatosis, collagen accumulation, ALT, AST, and Tbil. Increased Alb. Reduced BAX and increased Bcl-2.[136]
13 wksYiguanjian decoctionReduced ALT, α-SMA, and collagen deposition. Decreased F480, Alb, EGFP, PKM2, Ki-67, and AFP. [69]

Others

Ovariectomized rats8 wksCitrus unshiu peel extractDecreased hepatic lipid contents, AST, and ALT. Reduced hepatic lipid deposition. [137]

Porcine serum induced 16 wksRoots of Paeonia lactiflora and Astragalus membranaceusReduced liver damage and symptoms of liver fibrosis. Decreased HA, PC III, and Hyp. Restored SOD and GSH and inhibited lipid peroxidation. [138]

Schistosomiasis induced18 wksRadix Astragali, Salvia miltiorrhiza, and Angelica sinensisSome groups showed small reticulation and spot thickening in liver. Reduced ALT, albumin, and TBil. [139]
18 wksDanggui Buxue decoctionReversed fibrosis.[140]

Combination therapy

CCl4 + high lipid low protein diet6 wksDanggui Buxue decoctionReduced hepatic fatty degeneration and collagen accumulation. Decreased ALT, AST, and TBil. Reduced MDA, TG, Hyp content, and MMP-2/9 activities; increased SOD.[141]

CCl4, bile duct ligation, alcohol fed7 wksCoptidis rhizoma extract and bear bileReduced AST, Hyp, and Tbil. Decreased hepatic damage and fibrosis. Reduced peroxidative stress; increased SOD. [41]

CCl4 + high fat emulsion8 wksflavonoids from Litsea coreana LevlReduced ALT, AST, HA, laminin, procollagen III N-terminal peptide, procollagenase IV, and Hyp. Reduced hepatocyte degeneration, inflammatory cell infiltration, and collagen deposition. Suppressed α-SMA, collagen I, TGF-β1, and its receptor; increased PPARγ.[142]

DMN and CCl4 induced8 wks (DMN) + 10 wks (CCl4)Graptopetalum paraguayenseReduced ALT, AST, and Tbil. Suppressed collagen deposition. [143]

Content not specified in the paper.

Liver fibrogenesis results from excessive deposition of extracellular matrix and is part of a wound healing process triggered by activation of hepatic stellate cells. The process is accompanied by cell necrosis, apoptosis, and inflammation [27]. TGF-β1 plays an important role in liver injury, by regulating the inflammation process, hepatocyte apoptosis, and the transformation of hepatic stellate cells to myofibroblasts. Transformed myofibroblasts secrete matrix metalloproteinases (MMPs) that degrade the extracellular matrix of normal cells and further promote deposition of fibrillar collagens [26]. Due to the large number of myofibroblasts and collagen deposited in fibrotic regions, the hepatic expressions of α-SMA and type I/II collagen also increase significantly. Therefore, these biochemical markers as well as the inflammatory factors MCP-1 and TNFα frequently used to evaluate the effect of herbal medicines in experimental liver fibrosis models.

Cholestatic models where bile efflux is impeded through induction of obstructive bile duct injury are also frequently used to study the effect of herbal medicines on biliary fibrosis. After surgical bile duct ligation, animals develop periportal hepatocyte necrosis, liver failure, and fibrosis within one week. Inflammatory cell infiltration, hepatocyte apoptosis, and collagen deposition are observed in 4 weeks after bile duct ligation. This murine surgical model is often used because it is fast and reproducible, even though it can result in high mortality within a few weeks, an outcome that does not mirror the slow progression of the disease in humans [29]. In animals, undergoing bile duct ligation with administration of huangqi extract restored expression of TGF-β1, α-SMA, albumin, and CK7 markers [42]. Proposed mechanisms for the actions of herbal medicines on biliary fibrosis and models used to study the effects are summarized in Table 3.


ModelExperiment durationsHerbal medicinePathological and biochemical changes; mechanism involvedReference

Bile duct ligation3 daysAqueous extract from the root of Platycodon grandiflorumBlocked ALT and AST. Restored antioxidant enzymes. High dose showed lesser hepatocyte necrosis and inflammatory cell infiltration.[144]
2 weeksYang-Gan-wanReduced α1 (I) procollagen and α-SMA.[145]
2 weeksArtemisia capillarisReduced cholestatic markers and Hyp. Blocked liver injury and collagen deposition. Reduced α-SMA, PDGF, and TGF-β. [146]
28 daysGreen tea polyphenolReduced portosystemic shunting, fibrosis, intrahepatic angiogenesis, and mesenteric window vascular density. Decreased HIF-1α, VEGF, and phospho-Akt. [147]
4 weeksHuangqi decoctionReduced fibrosis degree. Increased Hyp content, CK7, and α-SMA. [148]
4 weeksHuangqi decoctionBlocked collagen deposition. Reduced ALT, Tbil, and Hyp. Inhibited TGF-β1, its receptors, SMAD3, and pERK1/2. [42]
7 weeksInchinko-toDecreased ALT and AST. Reduced TGF-β1 and α-SMA. [149]

4. Animal Models for HCC

Exposure to a variety of agents can lead to malignant transformation of hepatocytes, a multistep process characterized by recurrent genetic modifications. HCC is the most common type of primary hepatic cancer. Its development is related to several risk factors such as HBV and HCV infection, alcoholic liver disease, and NAFLD as well as exposure to environmental toxins such as aflatoxins and diethyl nitrosamine [43]. The most effective treatment for HCC is surgical removal of the affected liver tissue followed by liver transplantation. Because HCC is most commonly diagnosed in late stages where extrahepatic metastasis is often present, early therapeutic interventions have not been explored at length.

Not surprisingly, some of these disease-causing agents mentioned above have been used to establish animal models for HCC. There are HBV/HCV transgenic mice models as well as models in which genetic alterations are induced through either knocking out of tumour suppressor genes or overexpressing c-myc or TGF-α protooncogenes. These models retain some features of the multistep processes of HCC development in humans and are frequently used to delineate the role of specific genes in hepatocarcinogenesis as well as studying the outcome of host-tumour interactions on disease progression [44]. Choedon et al. [45] used a HBx15-c-myc mouse model for HCC, in which a truncated HBx allele is overexpressed together with c-myc, to show that Thapring, a traditional Tibetan medicine, restores liver function after 10 months of treatment with concomitant reduction in serum of SOD and VEGF levels. The significant antitumour effect of Thapring is presumably linked to increased expression of and the apoptosis [45].

Xenograft models in which tumour cell suspensions are implanted subcutaneously in mice have been extensively used to monitor tumour growth and effectiveness of new therapies. This is often the first-line model for anticancer agent screening in vivo. Recent study by the National Cancer Institute [46] showed up to 45% of anticancer agents confirmed to be clinically effective demonstrated cytotoxic effects in xenograft models of HCC. The H22 xenograft model that is established through injection with H22 cell lines is particularly attractive among other models, due to the relatively short induction time to develop solid tumours (approximately 2 weeks). Both subcutaneous and intraperitoneal injections can induce solid tumour formation [47] although the details for this cell line are not well described in the literature. Administration of Eupolyphaga sinensis inhibited H22 tumour growth by promoting secretion of TNF-α and IFN-γ as well as inducing apoptosis via increased Bax/Bcl-2 ratio and caspase-3 production [48]. Furthermore, coadministration of chemotherapeutic agents together with herbal medicines significantly increased the cytotoxic effects. Cao and colleagues [49] showed that coadministration of Fuzheng-Yiliu granules with low dose 5-fluorouracil potentiated antitumour activity and restored white blood cell count. Similarly, combination treatment of Chaiqiyigan granula with taxol had a stronger antitumour effect than that of taxol alone [50]. Using xenograft models, our research group has shown a potent antitumour effect of Coptidis rhizoma aqueous extract. The effect was associated with decreased levels of markers for cell proliferation and vessel density such as Ki67 and CD31 [51, 52].

Orthotopic models in which tumour cells are implanted directly into the organ of interest are considered more clinically relevant and better predictive models for drug efficacy because the emerging liver tumours better reflect the niche microenvironment. Although this model is more technically demanding, it recapitulates key events of the human disease, such as displacement of the normal cell population by tumour formation as well as the production of circulating metastatic tumour cells. Wang and colleagues [53] implanted HepG2 cells expressing red fluorescent protein into mouse livers and monitored tumour growth and metastasis by fluorescence imaging. Metastatic spread to pancreas and mediastinal lymph nodes was observed after 25 days of post-HepG2 implantation. Early treatment with Celastrus orbiculatus effectively blocked tumour growth. Similar to observations made in humans regarding the administration of chemotherapeutics in later stages of liver disease, the effect of Celastrus orbiculatus became less significant when the treatment was started after the tumours formed. In a previous study, we also implanted luciferase-tagged MHCC97L tumours into BalB/c nude mice in order to observe the effect of berberine, the major ingredient of Coptidis rhizome. The treatment effectively suppressed tumour growth and lung metastasis [54]. Some of the antitumour mechanisms of herbal medicines in liver cancer and the models used are summarized in Table 4.


ModelExperiment durationsHerbal medicineMechanism involvedReference

Implantation (ectopic)

H22 cells5 daysFuzheng-Yiliu granuleInduced cell apoptosis. Antitumour effect after combined with 5-FU. Restored decreased level of WBC and lymphocytes. [150]

H22 cells5 daysFuzheng-Yiliu granuleInduced apoptosis and CD3, CD4, and NK cells in peripheral blood. Increased serum IL-2 and TNFα levels.[49]

H22 cells5 daysTagalsinUpregulated wild type p53 and reduced Bcl-2. [151]

H22 cells10 daysGlycyrrhiza polysaccharideInduced apoptosis via P53/PI3K/AKT pathway. [152]

H22 cells10 daysMacrothelypteris viridifronsInhibited expressions of VEGF and CD34. [153, 154]

Hepa 1–6 cells14 daysYupingfeng powderReduced MDA, SOD activities in liver and lung.[155]

H22 cells14 daysEupolyphaga sinensis WalkerIncreased TNFα and IFN-γ and Bax/Bcl-2 ratio; activated caspase-3.[48]

H22 cells14 daysRadix Glycyrrhizae polysaccharideReduced Treg population and Foxp3. Increased serum Th1/Th2 cytokine. [156]

H22 cells15 daysToosendanin, from Melia toosendan Sieb. et Zucc.Reduced Bcl-2; increased Bax and Fas. [157]

Hep3B cells15 daysAndrographolide, from Andrographis paniculataReduced GSH; increased p-JNK, ASK1, MKK4, and c-Jun. [158]

S180 and H22 ascites cells15 daysDioscorea bulbifera L. rhizome[159]
20 daysJiedu Xiaozheng YinIncreased expressions of cyclin D and cyclin E.[49, 154]

Bel-7402 cells25 daysSulfated glycopeptide from Gekko swinhonis Guenther Inhibited bFGF production. [152, 160]

HepG2 cells3 wkspennogenyl saponins from rhizoma paridisDecreased Bcl-2 and increased Bax, cleaved caspase-8, and cleaved caspase-3; decreased ERK1/2 and Akt phosphorylation and increased p38 and JNK phosphorylation.[161]

SMMC-7721 cells4 wksLuteoloside from Gentiana macrophyllaDecreased ROS and NLRP3 inflammasome. Inactivated caspase-1 and IL-1β.[162]

MHCC97L cells4 wksCoptidis rhizoma aqueous extract Increased phosphorylation of eEF2. [48, 51]

MHCC97L cells4 wksHuanglian Jiedu decoctionInactivated eEF2, induced eEF2k and AMPK. [52]

MHCC97-H cells5 wksCorilaginActivated p-p53-p21(Cip1) -cdc2/cyclin B1.[156, 163]
Hedyotis diffusa Willd.Downregulated expressions of CDK2, cyclin E, and E2F1.[157, 164]
Jiedu Xiaozheng YinInhibited expressions of expression of Bmi1 and Wnt/beta-catenin. [158, 165]
Triptolide from Tripterygium wilfordiiIncreased ROS and caspase-3 activity; induced Bax and inhibit Bcl-2 after addition of 5-FU.[166]
Invigorating spleen and detoxification decoctionIncreased MHC I/MHC II expression.[167]
Livistona chinensis seedsInhibited angiogenesis via reduction of VEGF-A and VEGFR-2 and inhibited Notch, Dll4, and Jagged1.[168]

H22 cellsPinus massoniana bark extractInduced apoptosis via caspase-dependent pathways.[169]

Walker-256 cellsAitongxiao recipeReduced Bcl-2, survivin. [170]

H22 cellsXiaoai Jiedu RecipeReduced CCL3, CXCL2. [171]

Hep3B cellsPiper betle leaf extractsInduced MAPK-p73 pathway.[172]

H22 cellsCiji Hua’ai Baosheng granule formula[173]

H22 cellsOxytropis falcata[174]

H22 cellsCorn silk polysaccharidesImproved serum IL-2, IL-6, and TNFα levels. [175]

HepG2/EGFP cellsChaiqiyigan granulaIncreased Bax; reduced p53 and VEGF combined with taxol.[50]

Primary hepatic carcinomaBushen jianpi decoctionReduced VEGF expression.[176]

Implantation (orthotopic)

HepG2 cells expressing red fluorescent protein 25 daysCelastrus orbiculatus Thunb.Inhibited VEGF.[53]

MHCC97H cells35 daysBaicaleinReduced MMP-2, MMP-9, and u-PA; increased TIMP-1 and TIMP-2 expressions through ameliorated phosphorylation of ERK. [177]

Bel-7402 cells10 wksJianpijiedu FangIncreased PTEN, pFAK; and reduced PI3K. [178]

MHCC97L cellsSongyou yinRestored E-cadherin and reduced N-cadherin.[179]

Transgenic mice model

X15-myc mice12 mthThapring Reduced Bcl2 and overexpression of p21Waf1.[45]

Others

25% CCl4 injection, 8% ethanol solution as drinking fluid for 4 weeks, and 20 weeks of 0.5% CCl4-8% ethanol solution as drinking fluid 8 wksXiaochaihu decoctionReduced ONOO, MDA, VMA, LPS-P, and ALP-C; increased ALP-A.[180]

N-Methyl-N-nitrosourea28 wksAegle marmelos leaf extractDecreased IL-1β, IL-6, Bcl-2, and c-jun; increased p53 and IL-4. [181]

Content not specified in the paper.

5. Animal Models for Acute Liver Injury

There is a growing number of studies describing the effect of herbal medicines in acute liver failure. Acute liver failure is defined as a severe impairment of liver function within short duration and without a history of preexisting liver disease. Acute injury leading to liver failure may be less frequent in the clinic than other forms of liver failure and yet it is life-threatening [55]. In humans, drug-induced liver injury (e.g., from high doses of acetaminophen, antibiotics, or antituberculosis drugs) is the major cause of acute liver failure. Other leading causes are viral infections and accidental toxicity such as excessive alcohol consumption or mushroom poisoning as well as those caused by ischemic or metabolic disorders. Much current herbal medicine research focused on the hepatoprotective actions of herbal medicines in acute liver injury models induced by hepatotoxins, chemicals, or drugs. A list of hepatoprotective herbal medicines and related animal models is shown in Table 5. Most of these disease models are of short duration and include studies of acute biochemical parameters of liver injury such as liver enzyme levels (i.e., ALT and AST) and inflammatory markers as well as markers of organ health such as circulating albumin and total bilirubin. The histological changes for acute liver injury model are not as significant as the longer-term model. Yet, it could be identified through the induction of vacuole formation, infiltration of inflammatory cells, and hepatocytes necrosis and apoptosis. In many instances, the toxicity model is established either concurrently or following the administration of herbal medicines in order to determine an extract’s hepatoprotective effect. For example, after 15 days of treatment with an aqueous licorice extract, mice were given an acute oral dose of CCl4 and sacrificed after 8 hours. Licorice-pretreated mice had significantly lower circulating liver enzymes and increased antioxidant enzymes in the liver, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), glutathione reductase (GR), and glutathione S-transferase (GST), indicating a hepatotoxic preventive effect from the licorice extract [56]. In acetaminophen-induced hepatotoxicity, coadministration of Tournefortia sarmentosa reduced CAT, SOD, and GPx antioxidant enzyme levels indicating less liver stress and decreased levels of inflammatory factors (TNF-α, IL-1β, and IL-6) [57]. Most hepatoprotective herbal medicines reverse liver injury through attenuation of injury-associated oxidative stress and inflammation resulting in lower levels of circulating and total bilirubin. Because at the moment liver transplantation is the only cure for acute liver failure, there is an urgent need to further explore the hepatoprotective effect of herbal medicines.


ModelExperiment durationsHerbal medicinePathological and biochemical changes; mechanism involvedReference

Hepatotoxin induced

CCl4
induced
6 hrsTanreqing injectionReduced serum liver markers and inflammation.[182]
10 hrsLycium barbarum polysaccharidesReduced centrilobular necrosis and ALT level. Restored antioxidant enzymes, decreased NO and lipid peroxidation, attenuated hepatic inflammation, and reduced NF-κB activity. [183]
24 hrsTaohe Chengqi TangReduced ALT and AST. Showed normal lobular pattern with mild degree of fat deposition, necrosis, and inflammatory cell infiltrations. [184]
2 daysPunarnavashtak kwathShowed mild inflammation. Reduced ALT, AST, ALP, and SBRN. Increased glutathione, SOD, and catalase; decreased TBARS.[185]
2 daysSharbat-e-DeenarRestored glutathione, adenosine triphosphate, and G-6-Pase. [186]
2 daysMajoon-e-Dabeed-ul-wardDecreased serum enzymes, albumin, and urea. Reduced necrosis, hepatocyte regeneration, and inflammation. Reduced TBARS; restored glutathione, adenosine triphosphatase, and G-6-Pase in liver.[187]
5 daysJusticia schimperiana (Hochst. ex Nees) and Verbascum sinaiticum BenthShowed lesser injury and necrosis. Reduced ALT, AST, and ASP levels. [188]
5 daysPolydatinDecreased ALT and AST. Blocked degeneration, necrosis, and inflammatory cell infiltration. Reduced MDA, hepatic TNFα, IL-1β, COX-2, iNOS, and NF-κB; increased SOD, CAT, GPx, and TGFβ. [189]
5 daysMeconopsis integrifolia (Maxim.) FranchReduced ALT, AST, ALP, and TB. Demonstrated in vivo antioxidant activities.[190]
7 daysFlowers of Abelmoschus manihot (L.) MedicDecreased vacuole formation and hepatocellular necrosis. Reduced MDA, TNFα, IL-1β, and NO; increased GSH, SOD, GPx, CAT, and GST.[191]
7 daysRadix TetrastigmaDecreased ALT and AST. Improved hepatic lesion. Reduced MDA, Bax, and caspase-3; increased SOD.[192]
7 daysNaringeninBlocked necrosis and restored cellular arrangement. Decreased ALT, AST, and ALP. Reduced TNFα, iNOS, and COX-2; increased Nrf2 and HO-1. [193]
7 daysProsthechea michuacana W. E. HigginsReduced hepatic serum markers and total protein.[194]
8 daysSyzygium jambosRestored SGPT, SGOT, ALP, and total bilirubin. [195]
8 daysHeptapeptide from Carapax trionycisDecreased ALT and AST. Reduced injury, necrosis, and ballooning degeneration. Showed moderate hypertrophy. Reduced MDA and increased GSH-Px. [196]
10 daysEthanolic extract of Eruca sativa L.Showed reduced inflammation. Reduced MDA.[197]
10 daysFruit of Lagenaria brevifloraDecreased total bilirubin. Restored total protein, albumin, and globulin. Increased GSH and reduced MDA and H2O2.[198]
14 daysDendrobium huoshanenseReduced MDA; restored SOD, CAT, GPx, and GSH. [199]
15 daysSaponins in Radix TrichosanthisIncreased SOD and T-AOC; decreased MDA and LDH levels.[200]
15 daysLicorice aqueous extractReduced ALT, AST, and ALP. Increased total protein, albumin, and globulin. Improved liver SOD, CAT, GSH-Px, GR, GST activities, and GSH level; reduced MDA, liver hydroxyproline, and serum TNFα.[56]
3 wksEthanol extract of Grewia tenaxImproved hepatic necrosis. Reduced MDA. [201]
Ficus chlamydocarpaDecreased serum enzyme markers. Reduced GSH and liver MDA.[202]
Physalis peruvianaImproved oxidative stress markers. [203]
ShaoganduoganReduced transaminase; blocked degeneration and necrosis. Enhanced activity of Na+ -K+ ATPase, Ca2+ ATPase, and SOD; reduced MDA. [193, 204]

Chemical induced

D-galactosamine induced 14 daysXiao-Chai-Hu TangDecreased serum IL-6 and TNFα, FasmRNA, FasLmRNA, and Bax protein but increased Bcl-2. [205, 206]
22 daysLeucas aspera (LA) Willd.Showed reduced hepatocellular necrosis. Decreased ASAT, ALAT, ALP, TGL, TC, and TB. Elevated superoxide dismutase, catalase, glutathione peroxidase, and decreased lipid peroxidation levels in liver.[206]
22 daysBrassica nigra (L.) KochDecreased SGOT, SGPT, ALP, LDH, and γ-GT; increased total protein and albumin. Reduced hepatocyte degeneration. [207]
Extracts from processed Corni fructusDecreased ALT and AST. Improved liver damage. Increased SOD and reduced MDA.[205, 208]

LPS induced 6 hoursBai-Hu-TangNo observable cellular necrosis and inflammatory cell infiltration. Prevents increase of IL10, TNFα.[209]

D-galactosamine and LPS induced 1 hrEchinacoside, from Cistanche salsaDecreased ALT. Blocked hepatocyte apoptosis and improved histopathological changes. Reduced inflammatory markers.[210]
5 hrsXijiao Dihuang decoctionReduced ALT and AST. Blocked liver injury. [211]
36 hrsSanhuangyinchi decoctionDecreased ALT, TBIL, and PT. Inhibits caspase-3 activity.[212]
3 dFuzheng Huayu recipeBlocked hepatocyte apoptosis and reduced ALT and AST. Decreased MDA; improved SOD activity in liver. Decreased TNFα and TNFR1 protein expression. [213]
7 daysSanhuangyinchi decoctionReduced ALT, AST, TBIL, TP, and INR; increased FIB. Blocked the pathological changes. Increased SOD; decreased MDA and caspase-3 in liver.[214]
Qingxia therapyDecreased ALT, AST, and TBIL levels and hepatocyte necrosis and inflammatory cell infiltration. Reduced BAX, caspase-3 and increased Bcl-2.[215]

Drug induced

Paracetamol induced 24 hrsAndrographis paniculata and Swertia chirayitaReversed the elevated levels of GOT, GPT, ALP, and bilirubin. Restored liver architecture. Increased LPO; reduced SOD, catalase, GSH, and GPx. [216, 217]
7 dNigella sativa extractReduced liver enzymes and total bilirubin. Showed reduction of sinusoidal dilation, necrosis, and apoptosis.[217]

Acetaminophen induced 24 hrsTournefortia sarmentosa Lam.Reduced SGPT, SGOT, ALP, and bilirubin. Showed minimal fibrosis and periportal inflammation. Reduced inflammatory markers, MDA, and antioxidant enzyme levels.[57, 218]
8 daysCapparis sepiaria L.Reduced SGPT, SGOT, and ALP. [218]

Rifampicin induced in Wistar rat15 dEuphorbia fusiformis Buch.Reduced SGOT, SGPT, GGTP, ALP, and total bilirubin. Showed mild degeneration and necrosis.[219]

Antitubercular drug induced45 dHibiscus vitifolius Linn.Reduced AST, ALT, ALP, LDH, and total bilirubin and increased TC, TP, and albumin. Reduced necrosis and inflammatory cell infiltration. Increased catalase and SOD; reduced TARS. [220]

Others

Sodium arsenite induced in ratsOcimum basilicumReduced ALT and AST. [221]

Restrained stress induced in mouse 5 dMyelophil, an ethanol extract of Radix Astragali and Salviae RadixReduced ALT and AST. Reduced ROS and lipid peroxidation; restored liver catalase, glutathione reduced, and peroxidase; normalised IL1β. [222]

Dimethoate induced (insecticides) in guinea pigs21 dWithania somnifera extractReduced AST, ALT, and ALP. No observable pathological changes. [223]

Pyrogallol induced12 hrsCotinus coggygria Scop.Reversed AST, ALT, ALP, and total bilirubin. Reduced Akt and STAT3.[224]

Concanavalin-A 9 dSuaeda maritima (L.) DumortReduced serum liver markers. Restored liver architecture and showed less visible changes. [225]

Naphthalene induced30 dColeus aromaticus leaf extractShowed no necrosis and infiltration of inflammatory cells. Reduced AST, ALT, ACP, and ALP.[226]

Alpha-naphthylisothiocyanate (ANIT) induced5 dCalculus bovis SativusDecreased ALT, AST, ASP, and Tbil. Showed mild interlobular duct epithelial damages, lesser neutrophil cells infiltration, necrosis, and degeneration. Reduced MDA; increased hepatic SOD. [227]
9 dDanning tabletReduced ALT, AST, ALP, T-Bil, and D-Bil. Showed mild necrotic and degenerative changes with lesser neutrophil infiltration. Reduced hepatic MPO, GST, GSH, and LPO; increased SOD, Gpx, and CAT activities. [228]

Content not specified in the paper.

6. Discussion

6.1. Mechanisms Associated with Cytoprotective Effects of Herbal Medicines in Liver Diseases

Oxidative stress is one of the key drivers of liver disease pathogenesis. Damage by oxidative stress results when the natural balance between production and breakdown of reactive oxygen species (ROS) is disturbed. In NAFLD, beta-oxidation in mitochondria leads to disturbances in electron transport reactions and elevates ROS production [58]. ROS has been implicated in altered hepatocyte ploidy as well as the initiation of genomic changes that may further promote progression to HCC. In keeping with the reported deleterious effects of ROS on liver function, antioxidants contained in herbal remedies were shown to restore normal hepatocyte ploidy in NAFLD [59]. In the HFD induced NAFLD model, mice fed with excessive fat showed accelerated mitochondrial and peroxisomal fatty acids β-oxidation as well as increased microsomal fatty acid ω-oxidation [60]. Furthermore, ROS has also been implicated in the CCl4 model of liver fibrosis. CCl3 radicals resulting from administration of CCl4 interact with oxygen to produce highly reactive peroxy radicals that promote lipid peroxidation through removing hydrogen groups from unsaturated fatty acids [61]. One way that herbal medicine contributes to the amelioration of liver diseases is by improving antioxidant activity through chemical reduction of malondialdehyde (MDA) or by boosting glutathione S-transferase (GST) activity. High concentrations of the antioxidants, terpenes, and flavonoids in herbal medicines are likely responsible for the protective effect. In addition, herbal medicines increase the activity of many liver enzymes involved in ROS scavenging such as catalase, superoxide dismutase (SOD), and glutathione peroxide (GPx) as well as reducing ROS production via promoting formation of GSH, the antioxidant. Another instance of liver protection is associated with upregulation of UCP, a mitochondrial carrier [18, 62] involved in ROS metabolism.

Apart from oxidative stress, inflammation is another principal driver of liver disease. Persistent inflammation can be triggered through activation of resident macrophages and infiltration with immune cells from the blood stream following injury as well as the interaction of immune cells with surrounding liver tissues [27]. All these will lead to excessive secretion of proinflammatory cytokines TNF-α, IL-6, and IL-1β and further contribute to cell death. Furthermore, TGF-β secretion by activated monocytes was shown to stimulate hepatic stellate cells to increase collagen production [63]. These observations underscore the important role of inflammation-related host-cell interactions in liver disease. Many herbal medicines possess potent anti-inflammatory activities that attenuate cytokine or chemokine production. Although there are few studies reported on how herbal remedies regulate immune cells or hepatocytes to attenuate inflammation, several protein targets such as NF-κB, STAT3, and AMPK have been suggested. By inhibiting these pathways, some herbal remedies effectively protect the liver against inflammation-induced damage.

6.2. In Vitro Models for Liver Diseases

To date, herbal medicine research in liver diseases, particularly NAFLD/NASH and fibrosis, rarely employed in vitro cell models because of their limitation in mimicking clinical pathogenesis. Although immortalized cell lines and primary cultures have been used for herbal medicine studies, interactions between different cell types and the influence of extracellular matrix as well as other aspects in the niche microenvironment, which are significant disease-contributing factors in humans, cannot be replicated in cell culture models. Using nonalcoholic fatty liver disease as an example, interactions between adipocytes and hepatocytes regulate secretion of free fatty acids that further promotes the transition of hepatocytes and surrounding nonparenchymal cells towards the disease state. Thus, many factors need to be considered regarding whether a cell model of the liver disease is to produce valuable translational outcome. Still, due to the simplicity of studying molecular mechanisms of disease and the ease of obtaining drug leads, a lot of efforts have been directed at establishing or optimizing in vitro cell models for high throughput drug screening. Xu et al. [64] established a TGF-β1 fibrogenesis two-cell based model that allows efficient quantification of the effects of antifibrotic drugs on 2D matrix accumulation and 3D nodule formation. The model has been employed in kidney fibroblasts for the screening of Chinese medicine compounds and herb extracts for inflammation independent antifibrotic activity [65]. Another model established by Chen et al. [66] is the “scar-in-a-jar” model that improves current fibroplasia models by incorporating in situ optical bioimaging for cell and collagen quantitation. Yet, neither of these two models has been used at length to screen for antifibrotic herbal remedies. A substantial leap in our ability to investigate the use of herbal medicines in liver diseases is expected to occur following the establishment of new models or improvement of existing ones.

6.3. Future Perspectives

Due to the limitation of cell models to provide translatable solutions for clinical applications, animal models remain important for translating a promising herbal compound into the clinic. The preferred protocol is to firstly establish a genetically modified or diet fed mouse model that exhibits some key aspect of the human pathogenesis. In humans, liver disease pathogenesis requires years of genetic evolution and cellular dysfunction in which processes such as cross talk between parenchymal and nonparenchymal liver cells as well as involvement of immune cells occur. Thus, the effects of herbal medicines on these cellular processes need to be delineated in order to provide a better understanding of the processes involved and to facilitate better translational context for human studies.

Single animal model and short duration studies that aim to establish the effectiveness of any herbal medicine are relatively unpersuasive. Typically, such studies use a narrow spectrum of outcomes with limited measurement criteria that may be poorly translatable to human studies. Clearly, even in complex models, the complete pathogenesis of a specific liver disease is rarely recapitulated and therefore more than one animal model is needed to validate the effectiveness of an herbal remedy. Several herbal medicines mentioned above show a profound reversal of liver disease markers and need to be studied further. For example, the potent antifibrotic effect of Yi Guan Jian extract that has been extensively studied in the CCl4 model is attributed to the blockade of fibrogenesis due to bone marrow derived fibrogenic cells in the liver [6769]. However, this effect is oxidative stress-dependent and can only be partially reproduced by administration of CCl4 that severely damages the liver and does not reflect the conditions present in the human disease. Further studies using different rodent models should be implemented to eliminate the possibility that the therapeutic effect of this herbal remedy is due to model-specific bias.

Although preclinical studies have mostly used mouse models, alternative models based on zebrafish are now emerging. With a sequenced genome, the zebrafish has recently emerged as a robust vertebrate model for a variety of human diseases, including the liver diseases [70, 71]. Zebrafish is amenable to unparalleled ease of embryonic manipulation and adaptability for high throughput screens. Furthermore, the physiological similarities between zebrafish and the humans provide a strong rationale for direct drug discovery on zebrafish embryos with translational potential [72, 73]. Zebrafish studies further benefit from a vast array of newly developed research tools such as genome-wide ENU mutagenesis, transgenesis, and genome editing/gene knockout by ZFN, TALENs, and CRISPR. These tools are readily available to create gene-modified models of liver diseases that can be applied to large scale screens of herbal compounds/extract as well as mechanistic studies of disease progression [74].

Recently, zebrafish has been actively explored as models for NAFLD and/or steatosis using dietary addition of fructose [75] or by introducing a slc7a3a mutation to knockout genes involved in NO-AMPK-PPAR-γ signalling pathway [76]. ENU-mutagenized zebrafish larvae have also been used for genetic screens aimed at identifying novel genes that contribute to NAFLD/NASH phenotype [77]. Studies towards the development of zebrafish hepatic fibrosis model showed that administration of diethylnitrosamine resulted in 80% of treated zebrafish developing liver fibrosis after 6 weeks of treatment [78]. In an effort to build spontaneous hepatic cancer models in zebrafish, either constitutively or by using chemical inducers of liver-specific promoters, the Gong team has achieved targeted liver overexpression of a number of oncogenes such as Kras (V12) and Myc [7982]. The Stainier lab has also developed a hepatocyte-specific activated β-catenin model in which 78% of transgenic zebrafish developed hepatocellular carcinoma by 6 months of age [83]. It is anticipated that these unique genetic zebrafish liver disease models and others will lead to further mechanistic studies as well as large-scale screens of herbal medicines in the near future.

7. Conclusion

Herbal medicines contain a wealth of empirical pharmacological outcomes distilled over centuries of practice but more research efforts should be tried in identifying the active medicinal ingredients. This rediscovery or modernization of traditional Chinese medicine holds great promise for new active compounds with cytoprotective, disease-arresting, and curative properties. There are still many challenges to establish relevant animal models for studying the efficacy of the promising compounds outlined in this review. The process of animal modelling is crucial to generate valuable translatable outcomes. Currently, no single model exists that completely recapitulates the entire pathological progression of NAFLD, NASH, cirrhosis, HCC, or acute liver injury. Therefore, data obtained from one model needs to be validated and studied further in other models. It is anticipated that, by employing new genome editing technologies such as CRISPR, more faithful animal models of liver disease will emerge that will push the current boundaries of herbal medicine research. While rodent models remain paramount for the study of drug efficacy, mechanism of action, and toxicity, new emerging zebrafish models assisted by a host of recent technologies hold great promises for high throughput screens of new bioactive compounds in herbal medicines.

Conflict of Interests

The authors declare no conflict of interests.

Authors’ Contribution

Hor-Yue Tan and Serban San-Marina drafted the paper. All authors revised and commented on the paper and discussed the paper. Xiao-Yan Wen and Yibin Feng conceived, designed, revised, and finalized the paper.

Acknowledgments

This research was partially supported by the Research Council of the University of Hong Kong (Project codes: 104002889 and 104003422; Yibin Feng), Wong’s Donation (Project code: 200006276; Yibin Feng), the donation of Gaia Family Trust, New Zealand (Project code: 200007008; Yibin Feng), Canada Foundation for Innovation (Xiao-Yan Wen), and Brain Canada Foundation (Xiao-Yan Wen).

References

  1. Y. Liu, C. Meyer, C. Xu et al., “Animal models of chronic liver diseases,” The American Journal of Physiology—Gastrointestinal and Liver Physiology, vol. 304, no. 5, pp. G449–G468, 2013. View at: Publisher Site | Google Scholar
  2. Z. M. Younossi, M. Stepanova, M. Afendy et al., “Changes in the prevalence of the most common causes of chronic liver diseases in the United States from 1988 to 2008,” Clinical Gastroenterology and Hepatology, vol. 9, no. 6, pp. 524.e1–530.e1, 2011. View at: Publisher Site | Google Scholar
  3. G. Vernon, A. Baranova, and Z. M. Younossi, “Systematic review: the epidemiology and natural history of non-alcoholic fatty liver disease and non-alcoholic steatohepatitis in adults,” Alimentary Pharmacology and Therapeutics, vol. 34, no. 3, pp. 274–285, 2011. View at: Publisher Site | Google Scholar
  4. J. K. Dyson, Q. M. Anstee, and S. McPherson, “Non-alcoholic fatty liver disease: a practical approach to treatment,” Frontline Gastroenterology, vol. 5, no. 4, pp. 277–286, 2014. View at: Publisher Site | Google Scholar
  5. A. A. Mokdad, A. D. Lopez, S. Shahraz et al., “Liver cirrhosis mortality in 187 countries between 1980 and 2010: a systematic analysis,” BMC Medicine, vol. 12, article 145, 2014. View at: Publisher Site | Google Scholar
  6. D. Schuppan and N. H. Afdhal, “Liver cirrhosis,” The Lancet, vol. 371, no. 9615, pp. 838–851, 2008. View at: Publisher Site | Google Scholar
  7. A. C. Society, “Survival rates for liver cancer,” 2015, http://www.cancer.org/cancer/livercancer/detailedguide/liver-cancer-survival-rates. View at: Google Scholar
  8. M. W. Fried, V. J. Navarro, N. Afdhal et al., “Effect of silymarin (milk thistle) on liver disease in patients with chronic hepatitis C unsuccessfully treated with interferon therapy: a randomized controlled trial,” The Journal of the American Medical Association, vol. 308, no. 3, pp. 274–282, 2012. View at: Publisher Site | Google Scholar
  9. N.i.o. Health, “Treatment of Liver Cirrhosis Due to Hepatitis B Virus With Fuzheng Huayu and Entecavir,” 2014, https://clinicaltrials.gov/ct2/show/study/NCT02241590. View at: Google Scholar
  10. Q. Xu, Y. Feng, P. Duez, B. M. Hendry, and P. J. Hylands, “The hunt for antifibrotic and profibrotic botanicals,” Science, vol. 346, no. 6216, supplement, pp. S19–S20, 2014. View at: Google Scholar
  11. L. Hebbard and J. George, “Animal models of nonalcoholic fatty liver disease,” Nature Reviews Gastroenterology and Hepatology, vol. 8, no. 1, pp. 34–44, 2011. View at: Publisher Site | Google Scholar
  12. Y. Takahashi, Y. Soejima, and T. Fukusato, “Animal models of nonalcoholic fatty liver disease/ nonalcoholic steatohepatitis,” World Journal of Gastroenterology, vol. 18, no. 19, pp. 2300–2308, 2012. View at: Publisher Site | Google Scholar
  13. D. E. Kleiner, E. M. Brunt, M. Van Natta et al., “Design and validation of a histological scoring system for nonalcoholic fatty liver disease,” Hepatology, vol. 41, no. 6, pp. 1313–1321, 2005. View at: Publisher Site | Google Scholar
  14. X. Sheng, M. Wang, M. Lu, B. Xi, H. Sheng, and Y. Q. Zang, “Rhein ameliorates fatty liver disease through negative energy balance, hepatic lipogenic regulation, and immunomodulation in diet-induced obese mice,” The American Journal of Physiology—Endocrinology and Metabolism, vol. 300, no. 5, pp. E886–E893, 2011. View at: Publisher Site | Google Scholar
  15. J. Xiao, F. Xing, J. Huo et al., “Lycium barbarum polysaccharides therapeutically improve hepatic functions in non-alcoholic steatohepatitis rats and cellular steatosis model,” Scientific Reports, vol. 4, article 5587, 2014. View at: Publisher Site | Google Scholar
  16. H. Zheng, J. Zhao, Y. Liu, Y. Zheng, J. Wu, and Z. Hong, “Effect of total alkaloids of Rubus alceaefolius on oxidative stress in rats with non-alcoholic fatty liver disease,” Zhongguo Zhong Yao Za Zhi, vol. 36, no. 17, pp. 2383–2387, 2011. View at: Publisher Site | Google Scholar
  17. Y.-H. Jia, R.-Q. Wang, H.-M. Mi et al., “Fuzheng Huayu recipe prevents nutritional fibrosing steatohepatitis in mice,” Lipids in Health and Disease, vol. 11, article 45, 2012. View at: Publisher Site | Google Scholar
  18. Z.-M. Mao, H.-Y. Song, L.-L. Yang et al., “Effects of the mixture of Swertia pseudochmensis Hara and Silybum marianum Gaertn extracts on CCl4-induced liver injury in rats with non-alcoholic fatty liver disease,” Zhong Xi Yi Jie He Xue Bao, vol. 10, no. 2, pp. 193–199, 2012. View at: Publisher Site | Google Scholar
  19. Y.-Y. Wei, D. Yan, A. Japar, S.-S. Qu, A. A. Haji, and K. Parhat, “Effects of pomegranate flowers polyphenols on liver PON expression of diabetes combining non-alcoholic fat liver diseases rats,” Yao Xue Xue Bao, vol. 48, no. 1, pp. 71–76, 2013. View at: Google Scholar
  20. Y.-L. Ma, T. Li, B.-B. Wang et al., “Protection of huanglian jiedu decoction on livers of hyperlipidemia mice,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 33, no. 8, pp. 1107–1111, 2013. View at: Google Scholar
  21. Y. Takahashi, Y. Soejima, A. Kumagai, M. Watanabe, H. Uozaki, and T. Fukusato, “Inhibitory effects of Japanese herbal medicines sho-saiko-to and juzen-taiho-to on nonalcoholic steatohepatitis in mice,” PLoS ONE, vol. 9, no. 1, Article ID e87279, 2014. View at: Publisher Site | Google Scholar
  22. X. Li, Y. Li, W. Yang et al., “SREBP-1c overexpression induces triglycerides accumulation through increasing lipid synthesis and decreasing lipid oxidation and VLDL assembly in bovine hepatocytes,” Journal of Steroid Biochemistry and Molecular Biology, vol. 143, pp. 174–182, 2014. View at: Publisher Site | Google Scholar
  23. M. S. Shafiei, S. Shetty, P. E. Scherer, and D. C. Rockey, “Adiponectin regulation of stellate cell activation via PPARgamma-dependent and -independent mechanisms,” The American Journal of Pathology, vol. 178, no. 6, pp. 2690–2699, 2011. View at: Publisher Site | Google Scholar
  24. M. Ono, M. Ogasawara, A. Hirose et al., “Bofutsushosan, a Japanese herbal (Kampo) medicine, attenuates progression of nonalcoholic steatohepatitis in mice,” Journal of Gastroenterology, vol. 49, no. 6, pp. 1065–1073, 2014. View at: Publisher Site | Google Scholar
  25. S.-Y. Yang, N.-J. Zhao, X.-J. Li, H.-J. Zhang, K.-J. Chen, and C.-D. Li, “Ping-tang recipe improves insulin resistance and attenuates hepatic steatosis in high-fat diet-induced obese rats,” Chinese Journal of Integrative Medicine, vol. 18, no. 4, pp. 262–268, 2012. View at: Publisher Site | Google Scholar
  26. H. Hayashi and T. Sakai, “Animal models for the study of liver fibrosis: new insights from knockout mouse models,” American Journal of Physiology—Gastrointestinal and Liver Physiology, vol. 300, no. 5, pp. G729–G738, 2011. View at: Publisher Site | Google Scholar
  27. C. Liedtke, T. Luedde, T. Sauerbruch et al., “Experimental liver fibrosis research: update on animal models, legal issues and translational aspects,” Fibrogenesis and Tissue Repair, vol. 6, no. 1, article 19, 2013. View at: Publisher Site | Google Scholar
  28. R. G. Gieling, A. D. Burt, and D. A. Mann, “Fibrosis and cirrhosis reversibility—molecular mechanisms,” Clinics in Liver Disease, vol. 12, no. 4, pp. 915–937, 2008. View at: Publisher Site | Google Scholar
  29. P. Starkel and I. A. Leclercq, “Animal models for the study of hepatic fibrosis,” Best Practice and Research: Clinical Gastroenterology, vol. 25, no. 2, pp. 319–333, 2011. View at: Publisher Site | Google Scholar
  30. A. Baghdasaryan, P. Fickert, A. Fuchsbichler et al., “Role of hepatic phospholipids in development of liver injury in Mdr2 (Abcb4) knockout mice,” Liver International, vol. 28, no. 7, pp. 948–958, 2008. View at: Publisher Site | Google Scholar
  31. S. Kanzler, A. W. Lohse, A. Keil et al., “TGF-β1 in liver fibrosis: an inducible transgenic mouse model to study liver fibrogenesis,” American Journal of Physiology—Gastrointestinal and Liver Physiology, vol. 276, no. 4, part 1, pp. G1059–G1068, 1999. View at: Google Scholar
  32. Y.-N. Zhou, M.-Y. Sun, Y.-P. Mu et al., “Xuefuzhuyu decoction inhibition of angiogenesis attenuates liver fibrosis induced by CCl4 in mice,” Journal of Ethnopharmacology, vol. 153, no. 3, pp. 659–666, 2014. View at: Publisher Site | Google Scholar
  33. X. Shen, S. Cheng, Y. Peng, H. Song, and H. Li, “Attenuation of early liver fibrosis by herbal compound ‘Diwu Yanggan’ through modulating the balance between epithelial-to-mesenchymal transition and mesenchymal-to-epithelial transition,” BMC Complementary and Alternative Medicine, vol. 14, article 418, 2014. View at: Publisher Site | Google Scholar
  34. T.-F. Lee, Y.-L. Lin, and Y.-T. Huang, “Protective effects of kaerophyllin against liver fibrogenesis in rats,” European Journal of Clinical Investigation, vol. 42, no. 6, pp. 607–616, 2012. View at: Publisher Site | Google Scholar
  35. Q.-L. Wang, Y.-Y. Tao, L. Shen, H.-Y. Cui, and C.-H. Liu, “Chinese herbal medicine Fuzheng Huayu recipe inhibits liver fibrosis by mediating the transforming growth factor-beta1/Smads signaling pathway,” Zhong Xi Yi Jie He Xue Bao, vol. 10, no. 5, pp. 561–568, 2012. View at: Publisher Site | Google Scholar
  36. Y. Y. Tao, X. C. Yan, T. Zhou, L. Shen, Z. L. Liu, and C. H. Liu, “Fuzheng Huayu recipe alleviates hepatic fibrosis via inhibiting TNF-α induced hepatocyte apoptosis,” BMC Complementary and Alternative Medicine, vol. 14, article 449, 2014. View at: Publisher Site | Google Scholar
  37. C. Luo, Z.-X. Chen, X.-H. Tan et al., “Therapeutic effects of Fuzhenghuayu decoction in a CCl4-induced liver cirrhosis rat model and on hepatic stellate cell activation,” Zhonghua Gan Zang Bing Za Zhi, vol. 21, no. 9, pp. 668–673, 2013. View at: Google Scholar
  38. L. Wang, X. Yan, Z. Zeng, J. Lv, P. Liu, and C. Liu, “Effect of fuzheng huayu recipe and huangqi tang on DMN-induced experimental liver cirrhosis in rats,” Zhongguo Zhong Yao Za Zhi, vol. 35, no. 13, pp. 1740–1744, 2010. View at: Publisher Site | Google Scholar
  39. S. Dong, Q. L. Chen, and S. B. Su, “Curative effects of Fuzheng Huayu on liver fibrosis and cirrhosis: a meta-analysis,” Evidence-Based Complementary and Alternative Medicine, vol. 2015, Article ID 125659, 11 pages, 2015. View at: Publisher Site | Google Scholar
  40. Y. Feng, N. Wang, X. Ye et al., “Hepatoprotective effect and its possible mechanism of Coptidis rhizoma aqueous extract on carbon tetrachloride-induced chronic liver hepatotoxicity in rats,” Journal of Ethnopharmacology, vol. 138, no. 3, pp. 683–690, 2011. View at: Publisher Site | Google Scholar
  41. N. Wang, Y. Feng, F. Cheung et al., “A comparative study on the hepatoprotective action of bear bile and coptidis rhizoma aqueous extract on experimental liver fibrosis in rats,” BMC Complementary and Alternative Medicine, vol. 12, article 239, 2012. View at: Publisher Site | Google Scholar
  42. J.-X. Du, M.-Y. Sun, G.-L. Du et al., “Ingredients of Huangqi decoction slow biliary fibrosis progression by inhibiting the activation of the transforming growth factor-beta signaling pathway,” BMC Complementary and Alternative Medicine, vol. 12, article 33, 2012. View at: Publisher Site | Google Scholar
  43. Y. Li, Z. Y. Tang, and J. X. Hou, “Hepatocellular carcinoma: insight from animal models,” Nature Reviews Gastroenterology & Hepatology, vol. 9, no. 1, pp. 32–43, 2012. View at: Publisher Site | Google Scholar
  44. L. Bakiri and E. F. Wagner, “Mouse models for liver cancer,” Molecular Oncology, vol. 7, no. 2, pp. 206–223, 2013. View at: Publisher Site | Google Scholar
  45. T. Choedon, D. Dolma, and V. Kumar, “Pro-apoptotic and anticancer properties of Thapring—a Tibetan herbal formulation,” Journal of Ethnopharmacology, vol. 137, no. 1, pp. 320–326, 2011. View at: Publisher Site | Google Scholar
  46. J. I. Johnson, S. Decker, D. Zaharevitz et al., “Relationships between drug activity in NCI preclinical in vitro and in vivo models and early clinical trials,” British Journal of Cancer, vol. 84, no. 10, pp. 1424–1431, 2001. View at: Publisher Site | Google Scholar
  47. J. Zhang, X. Wang, and H. Lu, “Amifostine increases cure rate of cisplatin on ascites hepatoma 22 via selectively protecting renal thioredoxin reductase,” Cancer Letters, vol. 260, no. 1-2, pp. 127–136, 2008. View at: Publisher Site | Google Scholar
  48. G.-F. Ge, C.-H. Yu, B. Yu, Z.-H. Shen, D.-L. Zhang, and Q.-F. Wu, “Antitumor effects and chemical compositions of Eupolyphaga sinensis Walker ethanol extract,” Journal of Ethnopharmacology, vol. 141, no. 1, pp. 178–182, 2012. View at: Publisher Site | Google Scholar
  49. Z.-Y. Cao, X.-Z. Chen, L.-M. Liao et al., “Fuzheng Yiliu Granule inhibits the growth of hepatocellular cancer by regulating immune function and inducing apoptosis in vivo and in vitro,” Chinese Journal of Integrative Medicine, vol. 17, no. 9, pp. 691–697, 2011. View at: Publisher Site | Google Scholar
  50. M. You, M. Luo, W. Liao, S. Hu, W. Xu, and L. Jing, “Chaiqiyigan granula enhances Taxol-induced growth inhibition of hepatocellular carcinoma xenografts in nude mice: an in vivo fluorescence imaging study,” Nan Fang Yi Ke Da Xue Xue Bao, vol. 32, no. 7, pp. 1042–1045, 2012. View at: Google Scholar
  51. H. Y. Tan, N. Wang, S.-W. Tsao, Z. Zhang, and Y. Feng, “Suppression of vascular endothelial growth factor via inactivation of eukaryotic elongation factor 2 by alkaloids in Coptidis rhizome in hepatocellular carcinoma,” Integrative Cancer Therapies, vol. 13, no. 5, pp. 425–434, 2014. View at: Publisher Site | Google Scholar
  52. N. Wang, Y. Feng, H. Y. Tan et al., “Inhibition of eukaryotic elongation factor-2 confers to tumor suppression by a herbal formulation Huanglian-Jiedu decoction in human hepatocellular carcinoma,” Journal of Ethnopharmacology, vol. 164, pp. 309–318, 2015. View at: Publisher Site | Google Scholar
  53. M. Wang, X. Zhang, X. Xiong et al., “Efficacy of the Chinese traditional medicinal herb Celastrus orbiculatus Thunb on human hepatocellular carcinoma in an orthothopic fluorescent nude mouse model,” Anticancer Research, vol. 32, no. 4, pp. 1213–1220, 2012. View at: Google Scholar
  54. C. M. Tsang, K. C. P. Cheung, Y. C. Cheung et al., “Berberine suppresses Id-1 expression and inhibits the growth and development of lung metastases in hepatocellular carcinoma,” Biochimica et Biophysica Acta (BBA)—Molecular Basis of Disease, vol. 1852, no. 3, pp. 541–551, 2015. View at: Publisher Site | Google Scholar
  55. W. Bernal and J. Wendon, “Acute liver failure,” The New England Journal of Medicine, vol. 369, no. 26, pp. 2525–2534, 2013. View at: Publisher Site | Google Scholar
  56. H. Z. Huo, B. Wang, Y. K. Liang, Y. Y. Bao, and Y. Gu, “Hepatoprotective and antioxidant effects of licorice extract against CCl4-induced oxidative damage in rats,” International Journal of Molecular Sciences, vol. 12, no. 10, pp. 6529–6543, 2011. View at: Publisher Site | Google Scholar
  57. C.-Y. Teng, Y.-L. Lai, H.-I. Huang, W.-H. Hsu, C.-C. Yang, and W.-H. Kuo, “Tournefortia sarmentosa extract attenuates acetaminophen-induced hepatotoxicity,” Pharmaceutical Biology, vol. 50, no. 3, pp. 291–396, 2012. View at: Publisher Site | Google Scholar
  58. K. Begriche, J. Massart, M.-A. Robin, F. Bonnet, and B. Fromenty, “Mitochondrial adaptations and dysfunctions in nonalcoholic fatty liver disease,” Hepatology, vol. 58, no. 4, pp. 1497–1507, 2013. View at: Publisher Site | Google Scholar
  59. G. Gentric, V. Maillet, V. Paradis et al., “Oxidative stress promotes pathologic polyploidization in nonalcoholic fatty liver disease,” The Journal of Clinical Investigation, vol. 125, no. 3, pp. 981–992, 2015. View at: Publisher Site | Google Scholar
  60. G. Poli, “Pathogenesis of liver fibrosis: role of oxidative stress,” Molecular Aspects of Medicine, vol. 21, no. 3, pp. 49–98, 2000. View at: Publisher Site | Google Scholar
  61. M. Bhadauria, S. K. Nirala, and S. Shukla, “Propolis protects CYP 2E1 enzymatic activity and oxidative stress induced by carbon tetrachloride,” Molecular and Cellular Biochemistry, vol. 302, no. 1-2, pp. 215–224, 2007. View at: Publisher Site | Google Scholar
  62. W. Li, Y. Li, Q. Wang, and Y. Yang, “Crude extracts from Lycium barbarum suppress SREBP-1c expression and prevent diet-induced fatty liver through AMPK activation,” BioMed Research International, vol. 2014, Article ID 196198, 10 pages, 2014. View at: Publisher Site | Google Scholar
  63. S. Weber, O. A. Gressner, R. Hall, F. Grünhage, and F. Lammert, “Genetic determinants in hepatic fibrosis: from experimental models to fibrogenic gene signatures in humans,” Clinics in Liver Disease, vol. 12, no. 4, pp. 747–757, 2008. View at: Publisher Site | Google Scholar
  64. Q. Xu, J. T. Norman, S. Shrivastav, J. Lucio-Cazana, and J. B. Kopp, “In vitro models of TGF-β-induced fibrosis suitable for high-throughput screening of antifibrotic agents,” American Journal of Physiology—Renal Physiology, vol. 293, no. 2, pp. F631–F640, 2007. View at: Publisher Site | Google Scholar
  65. Q. Hu, M. Noor, Y. F. Wong et al., “In vitro anti-fibrotic activities of herbal compounds and herbs,” Nephrology Dialysis Transplantation, vol. 24, no. 10, pp. 3033–3041, 2009. View at: Publisher Site | Google Scholar
  66. C. Z. C. Chen, Y. X. Peng, Z. B. Wang et al., “The Scar-in-a-Jar: studying potential antifibrotic compounds from the epigenetic to extracellular level in a single well,” British Journal of Pharmacology, vol. 158, no. 5, pp. 1196–1209, 2009. View at: Publisher Site | Google Scholar
  67. Q. Tao, X.-N. Wang, Y.-P. Mu et al., “Dynamic change of lipid peroxidation-related protein expression and the intervention effects of Yiguanjian decoction in a rat model of CCl4-induced liver fibrosis,” Zhonghua Gan Zang Bing Za Zhi, vol. 20, no. 2, pp. 116–121, 2012. View at: Google Scholar
  68. X.-N. Wang, Q. Tao, Q. Feng, J.-H. Peng, P. Liu, and Y.-Y. Hu, “Effects of Chinese herbal medicine Yiguanjian decoction on collagen metabolism of hepatic tissues in rats with CCI4-induced liver fibrosis,” Zhong Xi Yi Jie He Xue Bao, vol. 9, no. 6, pp. 651–657, 2011. View at: Publisher Site | Google Scholar
  69. X.-L. Wang, D.-W. Jia, H.-Y. Liu et al., “Effect of Yiguanjian decoction on cell differentiation and proliferation in CCl4-treated mice,” World Journal of Gastroenterology, vol. 18, no. 25, pp. 3235–3249, 2012. View at: Publisher Site | Google Scholar
  70. W. Goessling and K. C. Sadler, “Zebrafish: an important tool for liver disease research,” Gastroenterology, vol. 149, no. 6, pp. 1361–1377, 2015. View at: Publisher Site | Google Scholar
  71. Y. Asaoka, S. Terai, I. Sakaida, and H. Nishina, “The expanding role of fish models in understanding non-alcoholic fatty liver disease,” Disease Models and Mechanisms, vol. 6, no. 4, pp. 905–914, 2013. View at: Publisher Site | Google Scholar
  72. F. Miscevic, O. Rotstein, and X.-Y. Wen, “Advances in zebrafish high content and high throughput technologies,” Combinatorial Chemistry and High Throughput Screening, vol. 15, no. 7, pp. 515–521, 2012. View at: Publisher Site | Google Scholar
  73. J. Tat, M. Liu, and X.-Y. Wen, “Zebrafish cancer and metastasis models for in vivo drug discovery,” Drug Discovery Today: Technologies, vol. 10, no. 1, pp. e83–e89, 2013. View at: Publisher Site | Google Scholar
  74. G. K. Varshney and S. M. Burgess, “Mutagenesis and phenotyping resources in zebrafish for studying development and human disease,” Briefings in Functional Genomics, vol. 13, no. 2, pp. 82–94, 2014. View at: Publisher Site | Google Scholar
  75. V. Sapp, L. Gaffney, S. F. EauClaire, and R. P. Matthews, “Fructose leads to hepatic steatosis in zebrafish that is reversed by mechanistic target of rapamycin (mTOR) inhibition,” Hepatology, vol. 60, no. 5, pp. 1581–1592, 2014. View at: Publisher Site | Google Scholar
  76. Q. Gu, X. Yang, L. Lin et al., “Genetic ablation of solute carrier family 7a3a leads to hepatic steatosis in zebrafish during fasting,” Hepatology, vol. 60, no. 6, pp. 1929–1941, 2015. View at: Publisher Site | Google Scholar
  77. S. H. Kim, S. Y. Wu, J. I. Baek et al., “A post-developmental genetic screen for zebrafish models of inherited liver disease,” PLoS ONE, vol. 10, no. 5, Article ID e0125980, 2015. View at: Publisher Site | Google Scholar
  78. K. Wang, L. Liu, W. Dai, X. Chen, X. Zheng, and J. Hou, “Establishment of a hepatic fibrosis model induced by diethylnitrosamine in zebrafish,” Nan Fang Yi Ke Da Xue Xue Bao, vol. 34, no. 6, pp. 777–782, 2014. View at: Google Scholar
  79. A. T. Nguyen, A. Emelyanov, C. H. V. Koh et al., “A high level of liver-specific expression of oncogenic KrasV12 drives robust liver tumorigenesis in transgenic zebrafish,” Disease Models and Mechanisms, vol. 4, no. 6, pp. 801–813, 2011. View at: Publisher Site | Google Scholar
  80. A. T. Nguyen, A. Emelyanov, C. H. Koh, J. M. Spitsbergen, S. Parinov, and Z. Gong, “An inducible KrasV12 transgenic zebrafish model for liver tumorigenesis and chemical drug screening,” Disease Models & Mechanisms, vol. 5, no. 1, pp. 63–72, 2012. View at: Publisher Site | Google Scholar
  81. Z. Li, W. Zheng, Z. Wang et al., “A transgenic zebrafish liver tumor model with inducible Myc expression reveals conserved Myc signatures with mammalian liver tumors,” Disease Models and Mechanisms, vol. 6, no. 2, pp. 414–423, 2013. View at: Publisher Site | Google Scholar
  82. L. Sun, A. T. Nguyen, J. M. Spitsbergen, and Z. Gong, “Myc-induced liver tumors in transgenic zebrafish can regress in tp53 null mutation,” PLoS ONE, vol. 10, no. 1, Article ID e0117249, 2015. View at: Publisher Site | Google Scholar
  83. K. J. Evason, M. T. Francisco, V. Juric et al., “Identification of chemical inhibitors of β-catenin-driven liver tumorigenesis in zebrafish,” PLoS Genetics, vol. 11, no. 7, Article ID e1005305, 2015. View at: Publisher Site | Google Scholar
  84. T. Liu, L.-L. Yang, L. Zhang, H.-Y. Song, D.-F. Li, and G. Ji, “Comparative study on the effects of different therapeutic methods in preventing and treating nonalcoholic fatty liver in rats,” Zhong Xi Yi Jie He Xue Bao, vol. 10, no. 10, pp. 1120–1126, 2012. View at: Publisher Site | Google Scholar
  85. Z. Yao, X.-C. Liu, and Y.-E. Gu, “Schisandra chinensis Baill, a Chinese medicinal herb, alleviates high-fat-diet-inducing non-alcoholic steatohepatitis in rats,” African Journal of Traditional, Complementary, and Alternative Medicines, vol. 11, no. 1, pp. 222–227, 2014. View at: Google Scholar
  86. Q. Peng, Q. Zhang, W. Xiao et al., “Protective effects of Sapindus mukorossi Gaertn against fatty liver disease induced by high fat diet in rats,” Biochemical and Biophysical Research Communications, vol. 450, no. 1, pp. 685–691, 2014. View at: Publisher Site | Google Scholar
  87. W. Tao, Z. Deqin, L. Yuhong et al., “Regulation effects on abnormal glucose and lipid metabolism of TZQ-F, a new kind of Traditional Chinese Medicine,” Journal of Ethnopharmacology, vol. 128, no. 3, pp. 575–582, 2010. View at: Publisher Site | Google Scholar
  88. L.-L. Yang, M. Wang, T. Liu et al., “Effects of Chinese herbal medicine Jiangzhi granule on expressions of liver X receptor α and sterol regulatory element-binding protein-1c in a rat model of non-alcoholic fatty liver disease,” Zhong Xi Yi Jie He Xue Bao, vol. 9, no. 9, pp. 998–1004, 2011. View at: Publisher Site | Google Scholar
  89. J. Liu, H. Zhang, B. Ji et al., “A diet formula of Puerariae radix, Lycium barbarum, Crataegus pinnatifida, and Polygonati rhizoma alleviates insulin resistance and hepatic steatosis in CD-1 mice and HepG2 cells,” Food and Function, vol. 5, no. 5, pp. 1038–1049, 2014. View at: Publisher Site | Google Scholar
  90. S. Chen, H. Zhou, M. Lin, R. Mi, and L. Li, “Decoction vs extracts-mixed solution: effect of yiqihuoxue formula on non-alcoholic fatty liver disease in rats,” Journal of Traditional Chinese Medicine, vol. 33, no. 4, pp. 513–517, 2013. View at: Publisher Site | Google Scholar
  91. Q.-H. Yang, S.-P. Hu, Y.-P. Zhang et al., “Effects of different therapeutic methods and typical recipes of Chinese medicine on activation of c-Jun N-terminal kinase in Kupffer cells of rats with fatty liver disease,” Chinese Journal of Integrative Medicine, vol. 18, no. 10, pp. 769–774, 2012. View at: Publisher Site | Google Scholar
  92. T.-Y. Lee, H.-H. Chang, W.-C. Lo, and H.-C. Lin, “Alleviation of hepatic oxidative stress by Chinese herbal medicine Yin-Chen-Hao-Tang in obese mice with steatosis,” International Journal of Molecular Medicine, vol. 25, no. 6, pp. 837–844, 2010. View at: Publisher Site | Google Scholar
  93. Y.-J. Kim, M.-S. Choi, Y. B. Park, S. R. Kim, M.-K. Lee, and U. J. Jung, “Garcinia cambogia attenuates diet-induced adiposity but exacerbates hepatic collagen accumulation and inflammation,” World Journal of Gastroenterology, vol. 19, no. 29, pp. 4689–4701, 2013. View at: Publisher Site | Google Scholar
  94. Q. Yang, Y. Xu, G. Feng et al., “p38 MAPK signal pathway involved in anti-inflammatory effect of Chaihu-Shugan-San and Shen-ling-bai-zhu-San on hepatocyte in non-alcoholic steatohepatitis rats,” African Journal of Traditional, Complementary, and Alternative Medicines, vol. 11, no. 1, pp. 213–221, 2014. View at: Google Scholar
  95. Q.-H. Yang, J. Huang, Y.-P. Zhang et al., “Effects of soothing liver and invigorating spleen recipes on the mRNA and protein expression of TLR4 in hepatic tissues of rats with NASH,” Zhong Yao Cai, vol. 36, no. 1, pp. 78–84, 2013. View at: Google Scholar
  96. Z.-F. Zhang, S.-H. Fan, Y.-L. Zheng et al., “Troxerutin improves hepatic lipid homeostasis by restoring NAD+-depletion-mediated dysfunction of lipin 1 signaling in high-fat diet-treated mice,” Biochemical Pharmacology, vol. 91, no. 1, pp. 74–86, 2014. View at: Publisher Site | Google Scholar
  97. L. Zhang, J. Xu, H. Song, Z. Yao, and G. Ji, “Extracts from Salvia-Nelumbinis naturalis alleviate hepatosteatosis via improving hepatic insulin sensitivity,” Journal of Translational Medicine, vol. 12, no. 1, article 236, 2014. View at: Publisher Site | Google Scholar
  98. Y. Ma, J. Zhao, S. Yang, and Y. Jia, “Cigu Xiaozhi pills's influence on lipid peroxidation and TNF-α expression in liver tissues of rats with nonalcoholic steatohepatitis,” Zhongguo Zhong Yao Za Zhi, vol. 35, no. 10, pp. 1292–1297, 2010. View at: Publisher Site | Google Scholar
  99. H.-Y. Song, Z.-M. Mao, L.-L. Yang et al., “Dangfei liganning capsules attenuate the susceptibility of rat nonalcoholic fatty liver to carbon tetrachloride toxicity,” Journal of Traditional Chinese Medicine, vol. 31, no. 4, pp. 327–333, 2011. View at: Publisher Site | Google Scholar
  100. Y.-J. Xu, Q.-H. Yang, L. Han et al., “Effects of soothing liver and invigorating spleen recipes on SREBP-1c, SCD-1 mRNA and proteins expression in hepatocytes of NAFLD rats,” Zhong Yao Cai, vol. 37, no. 1, pp. 80–86, 2014. View at: Google Scholar
  101. S. Takagi, T. Miura, E. Ishihara, T. Ishida, and Y. Chinzei, “Effect of corosolic acid on dietary hypercholesterolemia and hepatic steatosis in KK-Ay diabetic mice,” Biomedical Research, vol. 31, no. 4, pp. 213–218, 2010. View at: Publisher Site | Google Scholar
  102. Y. Hirotani, A. Doi, T. Takahashi, H. Umezawa, Y. Urashima, and M. Myotoku, “Protective effects of the herbal medicine goshajinkigan in a rat model of non-alcoholic fatty liver disease,” Biomedical Research, vol. 33, no. 6, pp. 373–376, 2012. View at: Publisher Site | Google Scholar
  103. Y. Xie, H.-P. Hao, H. Wang, Z.-X. Wang, and G.-J. Wang, “Reversing effects of silybin on TAA-induced hepatic CYP3A dysfunction through PXR regulation,” Chinese Journal of Natural Medicines, vol. 11, no. 6, pp. 645–652, 2013. View at: Publisher Site | Google Scholar
  104. D. Abdulaziz Bardi, M. F. Halabi, N. A. Abdullah, E. Rouhollahi, M. Hajrezaie, and M. A. Abdulla, “In vivo evaluation of ethanolic extract of Zingiber officinale rhizomes for its protective effect against liver cirrhosis,” BioMed Research International, vol. 2013, Article ID 918460, 10 pages, 2013. View at: Publisher Site | Google Scholar
  105. Y.-H. Paik, Y. J. Yoon, H. C. Lee et al., “Antifibrotic effects of magnesium lithospermate B on hepatic stellate cells and thioacetamide-induced cirrhotic rats,” Experimental and Molecular Medicine, vol. 43, no. 6, pp. 341–349, 2011. View at: Publisher Site | Google Scholar
  106. K.-G. Kwak, J.-H. Wang, J.-W. Shin, D.-S. Lee, and C.-G. Son, “A traditional formula, Chunggan extract, attenuates thioacetamide-induced hepatofibrosis via GSH system in rats,” Human & Experimental Toxicology, vol. 30, no. 9, pp. 1322–1332, 2011. View at: Publisher Site | Google Scholar
  107. S.-C. Chien, W.-C. Chang, P.-H. Lin et al., “A Chinese herbal medicine, Jia-Wei-Xiao-Yao-San, prevents dimethylnitrosamine-induced hepatic fibrosis in rats,” The Scientific World Journal, vol. 2014, Article ID 217525, 7 pages, 2014. View at: Publisher Site | Google Scholar
  108. C. Liu, P. Liu, and Q. Tao, “Recipe-syndrome correlation and pathogenesis mechanism of Yinchenhao Decoction in intervening dimethylnitrosamine induced liver cirrhosis progress in rats,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 30, no. 8, pp. 845–850, 2010. View at: Google Scholar
  109. C. Liu, G. Wang, G. Chen et al., “Huangqi decoction inhibits apoptosis and fibrosis, but promotes Kupffer cell activation in dimethylnitrosamine-induced rat liver fibrosis,” BMC Complementary and Alternative Medicine, vol. 12, article 51, 2012. View at: Publisher Site | Google Scholar
  110. Y.-Q. Bian, B.-B. Ning, H.-Y. Cao et al., “Formula-syndrome correlation study of three classical anti-jaundice formulas in inhibition of liver fibrosis induced by dimethylnitrosamine in rats,” Zhong Xi Yi Jie He Xue Bao, vol. 10, no. 12, pp. 1405–1412, 2012. View at: Publisher Site | Google Scholar
  111. X.-H. Tan, C.-Q. Li, S.-R. Zou et al., “Inhibitory effect of anluohuaxianwan on experimental hepatic fibrosis in rats,” Zhonghua Gan Zang Bing Za Zhi, vol. 18, no. 1, pp. 9–12, 2010. View at: Google Scholar
  112. X. Tong, G.-F. Chen, and Y. Lu, “Uniform designed research on the active ingredients assembling of huangqi decoction for inhibition of DMN-induced liver fibrosis,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 31, no. 10, pp. 1389–1393, 2011. View at: Google Scholar
  113. X. Zhang, B.-B. Ning, S. Ren et al., “Effects of Chinese herbal medicine Xiaopi Pill in preventing rats from dimethylnitrosamine-induced liver fibrosis,” Zhong Xi Yi Jie He Xue Bao, vol. 10, no. 11, pp. 1286–1292, 2012. View at: Publisher Site | Google Scholar
  114. C.-G. Liu, X.-L. Wang, X.-W. Du et al., “Metabolomic profiling for identification of potential biomarkers in the protective effects of modified sinisan against liver injury in dimethylnitrosamine treated rats,” Biological and Pharmaceutical Bulletin, vol. 36, no. 11, pp. 1700–1707, 2013. View at: Publisher Site | Google Scholar
  115. H.-J. Lin, J.-Y. Chen, C.-F. Lin et al., “Hepatoprotective effects of Yi Guan Jian, an herbal medicine, in rats with dimethylnitrosamine-induced liver fibrosis,” Journal of Ethnopharmacology, vol. 134, no. 3, pp. 953–960, 2011. View at: Publisher Site | Google Scholar
  116. İ. Bingül, C. Başaran-Küçükgergin, M. S. Tekkeşin, V. Olgaç, S. Doğru-Abbasoğlu, and M. Uysal, “Effect of blueberry pretreatment on diethylnitrosamine-induced oxidative stress and liver injury in rats,” Environmental Toxicology and Pharmacology, vol. 36, no. 2, pp. 529–538, 2013. View at: Publisher Site | Google Scholar
  117. R. Sferra, A. Vetuschi, V. Catitti et al., “Boswellia serrata and Salvia miltiorrhiza extracts reduce DMN-induced hepatic fibrosis in mice by TGF-beta1 downregulation,” European Review for Medical and Pharmacological Sciences, vol. 16, no. 11, pp. 1484–1498, 2012. View at: Google Scholar
  118. J.-Y. Chen, H.-L. Chen, J.-C. Cheng et al., “A Chinese herbal medicine, Gexia-Zhuyu Tang (GZT), prevents dimethylnitrosamine-induced liver fibrosis through inhibition of hepatic stellate cells proliferation,” Journal of Ethnopharmacology, vol. 142, no. 3, pp. 811–818, 2012. View at: Publisher Site | Google Scholar
  119. Y. Qian, X.-C. Fu, R. Hu, L.-M. Shen, and H.-B. Bai, “Effects of Corbrin Shugan capsule on dimethylnitrosamine-induced hepatic fibrosis in rats,” Zhejiang Da Xue Xue Bao Yi Xue Ban, vol. 42, no. 5, pp. 561–566, 2013. View at: Google Scholar
  120. Z. Zhao, H. Yu, Y. Peng et al., “Comparison of effect of formulas clearing away heat and promoting blood circulation on prevention and treatment of liver fibrosis in CCl4 mice,” Zhongguo Zhong Yao Za Zhi, vol. 37, no. 12, pp. 1804–1808, 2012. View at: Publisher Site | Google Scholar
  121. L. J. Zhang, M. Y. Sun, B. B. Ning et al., “Xiayuxue decoction ([symbols; see text]) attenuates hepatic stellate cell activation and sinusoidal endothelium defenestration in CCl4-induced fibrotic liver of mice,” Chinese Journal of Integrative Medicine, vol. 20, no. 7, pp. 516–523, 2014. View at: Publisher Site | Google Scholar
  122. X.-X. Wu, L.-M. Wu, J.-J. Fan et al., “Cortex Dictamni extract induces apoptosis of activated hepatic stellate cells via STAT1 and attenuates liver fibrosis in mice,” Journal of Ethnopharmacology, vol. 135, no. 1, pp. 173–178, 2011. View at: Publisher Site | Google Scholar
  123. H. Qiao, H. Han, D. Hong, Z. Ren, Y. Chen, and C. Zhou, “Protective effects of baicalin on carbon tetrachloride induced liver injury by activating PPARγ and inhibiting TGFβ1,” Pharmaceutical Biology, vol. 49, no. 1, pp. 38–45, 2011. View at: Publisher Site | Google Scholar
  124. G.-Y. Li, H.-Y. Gao, J. Huang, J. Lu, J.-K. Gu, and J.-H. Wang, “Hepatoprotective effect of Cichorium intybus L., a traditional Uighur medicine, against carbon tetrachloride-induced hepatic fibrosis in rats,” World Journal of Gastroenterology, vol. 20, no. 16, pp. 4753–4760, 2014. View at: Publisher Site | Google Scholar
  125. X.-P. Tian, Y.-Y. Yin, and X. Li, “Effects and mechanisms of acremoniumterricola milleretal mycelium on liver fibrosis induced by carbon tetrachloride in rats,” The American Journal of Chinese Medicine, vol. 39, no. 3, pp. 537–550, 2011. View at: Publisher Site | Google Scholar
  126. Z.-C. Wang, S. Yang, J.-J. Huang, S.-L. Chen, Q.-Q. Li, and Y. Li, “Effect of Rougan Huaqian granules combined with human mesenchymal stem cell transplantation on liver fibrosis in cirrhosis rats,” Asian Pacific Journal of Tropical Medicine, vol. 7, no. 7, pp. 576–581, 2014. View at: Publisher Site | Google Scholar
  127. F.-R. Yang, B.-W. Fang, and J.-S. Lou, “Effects of Fufang Biejia Ruangan Pills on hepatic fibrosis in vivo and in vitro,” World Journal of Gastroenterology, vol. 19, no. 32, pp. 5326–5333, 2013. View at: Publisher Site | Google Scholar
  128. C. Zhang, Y. Wang, H. Chen et al., “Protective effect of the herbal medicine Ganfukang against carbon tetrachloride induced liver fibrosis in rats,” Molecular Medicine Reports, vol. 8, no. 3, pp. 954–962, 2013. View at: Publisher Site | Google Scholar
  129. W. Li, Y. Wu, C. Zhu, Z. Wang, R. Gao, and Q. Wu, “Anti-fibrosis effects of Huisheng oral solution in CCl4-induced hepatic fibrosis in rat,” Indian Journal of Pharmacology, vol. 46, no. 2, pp. 216–221, 2014. View at: Publisher Site | Google Scholar
  130. A. N. B. Singab, N. A. Ayoub, E. N. Ali, and N. M. Mostafa, “Antioxidant and hepatoprotective activities of Egyptian moraceous plants against carbon tetrachloride-induced oxidative stress and liver damage in rats,” Pharmaceutical Biology, vol. 48, no. 11, pp. 1255–1264, 2010. View at: Publisher Site | Google Scholar
  131. H. B. Cai, X. G. Sun, Z. F. Liu et al., “Effects of dahuangzhechong pills on cytokines and mitogen activated protein kinase activation in rats with hepatic fibrosis,” Journal of Ethnopharmacology, vol. 132, no. 1, pp. 157–164, 2010. View at: Publisher Site | Google Scholar
  132. J.-X. Du, P. Liu, M.-Y. Sun et al., “Chinese herbal medicine Xiayuxue Decoction inhibits liver angiogenesis in rats with carbon tetrachloride-induced liver fibrosis,” Zhong Xi Yi Jie He Xue Bao, vol. 9, no. 8, pp. 878–887, 2011. View at: Publisher Site | Google Scholar
  133. D.-Z. Shen, Q. Tao, J.-X. Du et al., “Effects of Yiguanjian Decoction on liver cirrhosis formation a differential proteomics study in rats,” Zhong Xi Yi Jie He Xue Bao, vol. 8, no. 2, pp. 158–167, 2010. View at: Publisher Site | Google Scholar
  134. J.-C. Shu, L.-X. Chen, L. Deng et al., “Preliminary study on mechanism of therapeutic effect of Huganjiexian decoction on hepatic fibrosis,” Zhonghua Gan Zang Bing Za Zhi, vol. 18, no. 3, pp. 189–193, 2010. View at: Google Scholar
  135. Y.-P. Mu, X.-R. Chen, and Y.-F. Lu, “Effect of Xiaozheng Rongmu powder for the treatment of liver cirrhosis in rats,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 30, no. 10, pp. 1078–1083, 2010. View at: Google Scholar
  136. H. Jiang, J.-R. Gao, J.-F. Chen, and W.-B. Ji, “Effect of shuganjianpifang on the expression of BCL-2 and BAX in rats livers with hepatic fibrosis,” Zhong Yao Cai, vol. 36, no. 5, pp. 776–780, 2013. View at: Google Scholar
  137. D. W. Lim, Y. Lee, and Y. T. Kim, “Preventive effects of citrus unshiu peel extracts on bone and lipid metabolism in OVX rats,” Molecules, vol. 19, no. 1, pp. 783–794, 2014. View at: Publisher Site | Google Scholar
  138. W.-Y. Sun, L. Wang, H. Liu, X. Li, and W. Wei, “A standardized extract from Paeonia lactiflora and Astragalus membranaceus attenuates liver fibrosis induced by porcine serum in rats,” International Journal of Molecular Medicine, vol. 29, no. 3, pp. 491–498, 2012. View at: Publisher Site | Google Scholar
  139. X.-L. Liang and J.-Y. Yuan, “Effect of Chinese herbal compound on liver fibrosis in rabbits with schistosomiasis by B-ultrasound,” Asian Pacific Journal of Tropical Medicine, vol. 6, no. 8, pp. 658–662, 2013. View at: Publisher Site | Google Scholar
  140. P. Wang and Y.-Z. Liang, “Chemical composition and inhibitory effect on hepatic fibrosis of Danggui Buxue Decoction,” Fitoterapia, vol. 81, no. 7, pp. 793–798, 2010. View at: Publisher Site | Google Scholar
  141. Y. Chen, Q. Chen, J. Lu, F.-H. Li, Y.-Y. Tao, and C.-H. Liu, “Effects of Danggui Buxue Decoction (当归补血汤) on lipid peroxidation and MMP-2/9 activities of fibrotic liver in rats,” Chinese Journal of Integrative Medicine, vol. 15, no. 6, pp. 435–441, 2009. View at: Publisher Site | Google Scholar
  142. C. Huang, T. Ma, X. Meng et al., “Potential protective effects of a traditional Chinese herb, Litsea coreana Levl., on liver fibrosis in rats,” Journal of Pharmacy and Pharmacology, vol. 62, no. 2, pp. 223–230, 2010. View at: Publisher Site | Google Scholar
  143. L.-J. Su, C.-C. Chang, C.-H. Yang et al., “Graptopetalum paraguayense ameliorates chemical-induced rat hepatic fibrosis in vivo and inactivates stellate cells and Kupffer cells in vitro,” PLoS ONE, vol. 8, no. 1, Article ID e53988, 2013. View at: Publisher Site | Google Scholar
  144. T.-W. Kim, H.-K. Lee, I.-B. Song et al., “Protective effect of the aqueous extract from the root of Platycodon grandiflorum on cholestasis-induced hepatic injury in mice,” Pharmaceutical Biology, vol. 50, no. 12, pp. 1473–1478, 2012. View at: Publisher Site | Google Scholar
  145. M. D. Yang, Y.-M. Chiang, R. Higashiyama et al., “Rosmarinic acid and baicalin epigenetically derepress peroxisomal proliferator-activated receptor γ in hepatic stellate cells for their antifibrotic effect,” Hepatology, vol. 55, no. 4, pp. 1271–1281, 2012. View at: Publisher Site | Google Scholar
  146. J.-M. Han, H.-G. Kim, M.-K. Choi et al., “Artemisia capillaris extract protects against bile duct ligation-induced liver fibrosis in rats,” Experimental and Toxicologic Pathology, vol. 65, no. 6, pp. 837–844, 2013. View at: Publisher Site | Google Scholar
  147. S.-J. Hsu, S.-S. Wang, I.-F. Hsin et al., “Green tea polyphenol decreases the severity of portosystemic collaterals and mesenteric angiogenesis in rats with liver cirrhosis,” Clinical Science, vol. 126, no. 9, pp. 633–644, 2014. View at: Publisher Site | Google Scholar
  148. J.-X. Du, B.-F. Qiu, P. Liu, M.-Y. Sun, G.-F. Chen, and J. Liu, “Huangqi decoction inhibits cholangiocyte proliferation and transdifferentiation in cholestatic liver fibrosis induced by BDL in rats,” Zhonghua Gan Zang Bing Za Zhi, vol. 18, no. 1, pp. 13–18, 2010. View at: Publisher Site | Google Scholar
  149. T. Asakawa, M. Yagi, Y. Tanaka et al., “The herbal medicine Inchinko-to reduces hepatic fibrosis in cholestatic rats,” Pediatric Surgery International, vol. 28, no. 4, pp. 379–384, 2012. View at: Publisher Site | Google Scholar
  150. Z. Cao, L. Liao, X. Chen et al., “Enhancement of antitumor activity of low-dose 5-fluorouracil by combination with Fuzheng-Yiliu granules in hepatoma 22 tumor-bearing mice,” Integrative Cancer Therapies, vol. 12, no. 2, pp. 174–181, 2013. View at: Publisher Site | Google Scholar
  151. X. Q. Song, Y. L. Guo, B. G. Wang, S. J. Sun, and R. Y. Yao, “Effect of tagalsin on p53 and Bcl-2 expression in hepatoma H(22) tumor-bearing mice,” Zhonghua Zhong Liu Za Zhi, vol. 33, no. 7, pp. 499–503, 2011. View at: Google Scholar
  152. J. Chen, X. Jin, J. Chen, and C. Liu, “Glycyrrhiza polysaccharide induces apoptosis and inhibits proliferation of human hepatocellular carcinoma cells by blocking PI3K/AKT signal pathway,” Tumor Biology, vol. 34, no. 3, pp. 1381–1389, 2013. View at: Publisher Site | Google Scholar
  153. A. Wei, D. Zhou, J. Ruan, Y. Cai, C. Xiong, and G. Wu, “Anti-tumor and anti-angiogenic effects of Macrothelypteris viridifrons and its constituents by HPLC-DAD/MS analysis,” Journal of Ethnopharmacology, vol. 139, no. 2, pp. 373–380, 2012. View at: Publisher Site | Google Scholar
  154. Z. Cao, W. Lin, Z. Huang et al., “Ethyl acetate extraction from a Chinese herbal formula, Jiedu Xiaozheng Yin, inhibits the proliferation of hepatocellular carcinoma cells via induction of G0/G1 phase arrest in vivo and in vitro,” International Journal of Oncology, vol. 42, no. 1, pp. 202–210, 2013. View at: Publisher Site | Google Scholar
  155. L.-R. Zhang, Y. Tang, and G.-R. Jiang, “The protection of yupingfeng powder on cisplatin induced oxidative damage of organs in hepatocellular carcinoma mice,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 32, no. 5, pp. 647–651, 2012. View at: Google Scholar
  156. X. He, X. Li, B. Liu, L. Xu, H. Zhao, and A. Lu, “Down-regulation of Treg cells and up-regulation of TH1/TH2 cytokine ratio were induced by polysaccharide from Radix Glycyrrhizae in H22 hepatocarcinoma bearing mice,” Molecules, vol. 16, no. 10, pp. 8343–8352, 2011. View at: Publisher Site | Google Scholar
  157. Y. He, J. Wang, X. Liu et al., “Toosendanin inhibits hepatocellular carcinoma cells by inducing mitochondria-dependent apoptosis,” Planta Medica, vol. 76, no. 13, pp. 1447–1453, 2010. View at: Publisher Site | Google Scholar
  158. L. Ji, K. Shen, P. Jiang, G. Morahan, and Z. Wang, “Critical roles of cellular glutathione homeostasis and jnk activation in andrographolide-mediated apoptotic cell death in human hepatoma cells,” Molecular Carcinogenesis, vol. 50, no. 8, pp. 580–591, 2011. View at: Publisher Site | Google Scholar
  159. J.-M. Wang, L.-L. Ji, C. J. Banford-White et al., “Antitumor activity of Dioscorea bulbifera L. rhizome in vivo,” Fitoterapia, vol. 83, no. 2, pp. 388–394, 2012. View at: Publisher Site | Google Scholar
  160. S.-X. Zhang, C. Zhu, Y. Ba et al., “Gekko-sulfated glycopeptide inhibits tumor angiogenesis by targeting basic fibroblast growth factor,” The Journal of Biological Chemistry, vol. 287, no. 16, pp. 13206–13215, 2012. View at: Publisher Site | Google Scholar
  161. Y.-S. Chen, Y. He, C. Chen et al., “Growth inhibition by pennogenyl saponins from Rhizoma paridis on hepatoma xenografts in nude mice,” Steroids, vol. 83, pp. 39–44, 2014. View at: Publisher Site | Google Scholar
  162. S.-H. Fan, Y.-Y. Wang, J. Lu et al., “Luteoloside suppresses proliferation and metastasis of hepatocellular carcinoma cells by inhibition of NLRP3 inflammasome,” PLoS ONE, vol. 9, no. 2, Article ID e89961, 2014. View at: Publisher Site | Google Scholar
  163. Y. Ming, Z. Zheng, L. Chen et al., “Corilagin inhibits hepatocellular carcinoma cell proliferation by inducing G2/M phase arrest,” Cell Biology International, vol. 37, no. 10, pp. 1046–1054, 2013. View at: Publisher Site | Google Scholar
  164. X.-Z. Chen, Z.-Y. Cao, T.-S. Chen et al., “Water extract of Hedyotis Diffusa Willd suppresses proliferation of human HepG2 cells and potentiates the anticancer efficacy of low-dose 5-fluorouracil by inhibiting the CDK2-E2F1 pathway,” Oncology Reports, vol. 28, no. 2, pp. 742–748, 2012. View at: Publisher Site | Google Scholar
  165. X.-Z. Chen, Z.-Y. Cao, J.-N. Li et al., “Ethyl acetate extract from Jiedu Xiaozheng Yin inhibits the proliferation of human hepatocellular carcinoma cells by suppressing polycomb gene product Bmi1 and Wnt/β-catenin signaling,” Oncology Reports, vol. 32, no. 6, pp. 2710–2718, 2014. View at: Publisher Site | Google Scholar
  166. Y. Li and S. Hu, “Triptolide sensitizes liver cancer cell lines to chemotherapy in vitro and in vivo,” Panminerva Medica, vol. 56, no. 3, pp. 211–220, 2014. View at: Google Scholar
  167. Y.-L. Li, B.-G. Sun, T. Xiang, Z.-X. Chen, and S.-J. Zhang, “Effect of invigorating spleen and detoxification decoction on MHC I/MHC II in spleen-deficiency liver cancer rats survival,” Zhong Yao Cai, vol. 37, no. 3, pp. 454–460, 2014. View at: Google Scholar
  168. W. Lin, J. Zhao, Z. Cao et al., “Livistona chinensis seeds inhibit hepatocellular carcinoma angiogenesis in vivo via suppression of the Notch pathway,” Oncology Reports, vol. 31, no. 4, pp. 1723–1728, 2014. View at: Publisher Site | Google Scholar
  169. H. Ma, B. Liu, D. Feng et al., “Pinus massoniana bark extract selectively induces apoptosis in human hepatoma cells, possibly through caspase-dependent pathways,” International Journal of Molecular Medicine, vol. 25, no. 5, pp. 751–759, 2010. View at: Publisher Site | Google Scholar
  170. S.-J. Wang, A.-L. Wei, and Y.-Q. Zhang, “Aitongxiao recipe regulated survivin and Bcl- 2 in rats' transplanted hepatoma carcinoma cell,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 32, no. 12, pp. 1652–1657, 2012. View at: Google Scholar
  171. M.-H. Wu and L. Li, “Effect of xiaoai jiedu recipe on gene expression profiles in H22 tumor-bearing mice,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 33, no. 9, pp. 1232–1235, 2013. View at: Google Scholar
  172. P.-F. Wu, H.-C. Tseng, C.-C. Chyau, J.-H. Chen, and F.-P. Chou, “Piper betle leaf extracts induced human hepatocellular carcinoma Hep3B cell death via MAPKs regulating the p73 pathway in vitro and in vivo,” Food and Function, vol. 5, no. 12, pp. 3320–3328, 2014. View at: Publisher Site | Google Scholar
  173. S. Xi, R. Hong, J. Huang et al., “Effects of Ciji Hua'ai Baosheng Granule Formula (CHBGF) on life time, pathology, peripheral blood cells of tumor chemotherapymodelmouse with H22 hepatoma carcinomacells,” African Journal of Traditional, Complementary and Alternative Medicines, vol. 11, no. 4, pp. 94–100, 2014. View at: Publisher Site | Google Scholar
  174. G.-M. Yang, R. Yan, Z.-X. Wang, F.-F. Zhang, Y. Pan, and B.-C. Cai, “Antitumor effects of two extracts from Oxytropis falcata on hepatocellular carcinoma in vitro and in vivo,” Chinese Journal of Natural Medicines, vol. 11, no. 5, pp. 519–524, 2013. View at: Publisher Site | Google Scholar
  175. J. Yang, X. Li, Y. Xue, N. Wang, and W. Liu, “Anti-hepatoma activity and mechanism of corn silk polysaccharides in H22 tumor-bearing mice,” International Journal of Biological Macromolecules, vol. 64, pp. 276–280, 2014. View at: Publisher Site | Google Scholar
  176. Y. Zhong, C.-L. Luo, and Z. An-Jun, “Effect of bushen jianpi decoction and its disassemble recipes on tumor growth in mice with transplanted primary hepatic carcinoma,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 31, no. 2, pp. 213–217, 2011. View at: Google Scholar
  177. K. Chen, S. Zhang, Y. Ji et al., “Baicalein inhibits the invasion and metastatic capabilities of hepatocellular carcinoma cells via down-regulation of the ERK pathway,” PLoS ONE, vol. 8, no. 9, Article ID e72927, 2013. View at: Publisher Site | Google Scholar
  178. B. Sun, J. Meng, T. Xiang et al., “Jianpijiedu Fang improved survival of hepatocarcinoma mice by affecting phosphatase and tensin homolog, phosphoinositide 3-kinase, and focal adhesion kinase,” Journal of Traditional Chinese Medicine, vol. 33, no. 4, pp. 479–485, 2013. View at: Publisher Site | Google Scholar
  179. W. Xiong, Z.-Y. Tang, Z.-G. Ren et al., “Effects of the Chinese herbal extract Songyou Yin on the residual hepatocellular carcinoma after chemotherapy in nude mice,” Zhonghua Zhong Liu Za Zhi, vol. 35, no. 11, pp. 804–807, 2013. View at: Google Scholar
  180. X.-Q. Liu, X.-J. Hu, H.-X. Xu, and X.-Y. Zeng, “Xiaochaihu Decoction attenuates the vicious circle between the oxidative stress and the ALP inactivation through LPS-catecholamines interactions in gut, liver and brain during CCI4+ethanol-induced mouse HCC,” BMC Complementary and Alternative Medicine, vol. 13, article 375, 2013. View at: Publisher Site | Google Scholar
  181. S. Verma, T. Bahorun, R. K. Singh, O. I. Aruoma, and A. Kumar, “Effect of Aegle marmelos leaf extract on N-methyl N-nitrosourea-induced hepatocarcinogensis in Balb/c mice,” Pharmaceutical Biology, vol. 51, no. 10, pp. 1272–1281, 2013. View at: Publisher Site | Google Scholar
  182. Y. Lei, A.-M. Zhou, T. Guo, Y. Tan, Y.-Y. Tao, and C.-H. Liu, “Protective effect of Tanreqing injection on acute hepatic injury induced by CCl4 in rats,” Zhongguo Zhong Yao Za Zhi, vol. 38, no. 8, pp. 1226–1230, 2013. View at: Google Scholar
  183. J. Xiao, E. C. Liong, Y. P. Ching et al., “Lycium barbarum polysaccharides protect mice liver from carbon tetrachloride-induced oxidative stress and necroinflammation,” Journal of Ethnopharmacology, vol. 139, no. 2, pp. 462–470, 2012. View at: Publisher Site | Google Scholar
  184. T.-Y. Lai, Y.-J. Weng, W.-W. Kuo et al., “Taohe Chengqi Tang ameliorates acute liver injury induced by carbon tetrachloride in rats,” Zhong Xi Yi Jie He Xue Bao, vol. 8, no. 1, pp. 49–55, 2010. View at: Publisher Site | Google Scholar
  185. V. N. Shah, M. B. Shah, and P. A. Bhatt, “Hepatoprotective activity of punarnavashtak kwath, an Ayurvedic formulation, against CCl4-induced hepatotoxicity in rats and on the HepG2 cell line,” Pharmaceutical Biology, vol. 49, no. 4, pp. 408–415, 2011. View at: Publisher Site | Google Scholar
  186. A. K. Shakya, M. Saxena, N. Sharma, S. Shrivastava, and S. Shukla, “Hepatoprotective efficacy of sharbat-e-deenar against carbon tetrachloride-induced liver damage,” Journal of Environmental Pathology, Toxicology and Oncology, vol. 31, no. 2, pp. 131–141, 2012. View at: Publisher Site | Google Scholar
  187. A. K. Shakya, N. Sharma, M. Saxena, S. Shrivastava, and S. Shukla, “Evaluation of the antioxidant and hepatoprotective effect of Majoon-e-Dabeed-ul-ward against carbon tetrachloride induced liver injury,” Experimental and Toxicologic Pathology, vol. 64, no. 7-8, pp. 767–773, 2012. View at: Publisher Site | Google Scholar
  188. S. Umer, K. Asres, and C. Veeresham, “Hepatoprotective activities of two Ethiopian medicinal plants,” Pharmaceutical Biology, vol. 48, no. 4, pp. 461–468, 2010. View at: Publisher Site | Google Scholar
  189. H. Zhang, C.-H. Yu, Y.-P. Jiang et al., “Protective effects of polydatin from Polygonum cuspidatum against carbon tetrachloride-induced liver injury in mice,” PLoS ONE, vol. 7, no. 9, Article ID e46574, 2012. View at: Publisher Site | Google Scholar
  190. G. Zhou, Y. Chen, S. Liu, X. Yao, and Y. Wang, “In vitro and in vivo hepatoprotective and antioxidant activity of ethanolic extract from Meconopsis integrifolia (Maxim.) Franch,” Journal of Ethnopharmacology, vol. 148, no. 2, pp. 664–670, 2013. View at: Publisher Site | Google Scholar
  191. G. Ai, Q. Liu, W. Hua, Z. Huang, and D. Wang, “Hepatoprotective evaluation of the total flavonoids extracted from flowers of Abelmoschus manihot (L.) Medic: in vitro and in vivo studies,” Journal of Ethnopharmacology, vol. 146, no. 3, pp. 794–802, 2013. View at: Publisher Site | Google Scholar
  192. G. Cao, Q. Li, X. Chen, H. Cai, and S. Tu, “Hepatoprotective effect of superfine particles of herbal medicine against CCl4-induced acute liver damage in rats,” BioMed Research International, vol. 2014, Article ID 934732, 6 pages, 2014. View at: Publisher Site | Google Scholar
  193. M. A. Esmaeili and M. Alilou, “Naringenin attenuates CCl4-induced hepatic inflammation by the activation of an Nrf2-mediated pathway in rats,” Clinical and Experimental Pharmacology and Physiology, vol. 41, no. 6, pp. 416–422, 2014. View at: Publisher Site | Google Scholar
  194. R. M. P. Gutierrez, I. A. Sosa, C. H. Vadillo, and T. C. Victoria, “Effect of flavonoids from Prosthechea michuacana on carbon tetrachloride induced acute hepatotoxicity in mice,” Pharmaceutical Biology, vol. 49, no. 11, pp. 1121–1127, 2011. View at: Publisher Site | Google Scholar
  195. M. R. Islam, M. S. Parvin, and M. E. Islam, “Antioxidant and hepatoprotective activity of an ethanol extract of Syzygium jambos (L.) leaves,” Drug Discoveries & Therapeutics, vol. 6, no. 4, pp. 205–211, 2012. View at: Publisher Site | Google Scholar
  196. P. Wang, Y. Zhang, Y. An et al., “Protection of a new heptapeptide from carapax trionycis against carbon tetrachloride-induced acute liver injury in mice,” Chemical and Pharmaceutical Bulletin, vol. 61, no. 11, pp. 1130–1135, 2013. View at: Publisher Site | Google Scholar
  197. S. Alqasoumi, “Carbon tetrachloride-induced hepatotoxicity: protective effect of ‘Rocket’ Eruca sativa L. in rats,” American Journal of Chinese Medicine, vol. 38, no. 1, pp. 75–88, 2010. View at: Publisher Site | Google Scholar
  198. A. B. Saba, O. M. Onakoya, and A. A. Oyagbemi, “Hepatoprotective and in vivo antioxidant activities of ethanolic extract of whole fruit of Lagenaria breviflora,” Journal of Basic and Clinical Physiology and Pharmacology, vol. 23, no. 1, pp. 27–32, 2012. View at: Publisher Site | Google Scholar
  199. C.-C. Tian, X.-Q. Zha, L.-H. Pan, and J.-P. Luo, “Structural characterization and antioxidant activity of a low-molecular polysaccharide from Dendrobium huoshanense,” Fitoterapia, vol. 91, pp. 247–255, 2013. View at: Publisher Site | Google Scholar
  200. Y. Chen, Y. Miao, L. Huang et al., “Antioxidant activities of saponins extracted from Radix Trichosanthis: an in vivo and in vitro evaluation,” BMC Complementary and Alternative Medicine, vol. 14, article 86, 2014. View at: Publisher Site | Google Scholar
  201. M. S. Al-Said, R. A. Mothana, M. O. Al-Sohaibani, and S. Rafatullah, “Ameliorative effect of Grewia tenax (Forssk) fiori fruit extract on CCl4-induced oxidative stress and hepatotoxicity in rats,” Journal of Food Science, vol. 76, no. 9, pp. T200–T206, 2011. View at: Publisher Site | Google Scholar
  202. J. H. Donfack, C. C. F. Simo, B. Ngameni et al., “Antihepatotoxic and antioxidant activities of methanol extract and isolated compounds from Ficus chlamydocarpa,” Natural Product Communications, vol. 5, no. 10, pp. 1607–1612, 2010. View at: Google Scholar
  203. S. E. El-Gengaihi, M. A. Hamed, A. E.-R. M. Khalaf-Allah, and M. A. Mohammed, “Golden berry juice attenuates the severity of hepatorenal injury,” Journal of Dietary Supplements, vol. 10, no. 4, pp. 357–369, 2013. View at: Publisher Site | Google Scholar
  204. J. Song, J. Zhao, X. Wang, Y. Dai, Z. Deng, and J. Yi, “Protective effects of Shaoganduogan on hepatocyte mitochondria in subacute liver injury rat induced by carbon tetrachloride,” Zhongguo Zhong Yao Za Zhi, vol. 36, no. 7, pp. 931–934, 2011. View at: Publisher Site | Google Scholar
  205. Y.-X. Zhou, Y.-Q. Qiu, L.-Q. Xu, J. Guo, and L.-J. Li, “Xiao-Chai-Hu Tang in treating model mice with D-galactosamine-induced liver injury,” African Journal of Traditional, Complementary and Alternative Medicines, vol. 9, no. 3, pp. 405–411, 2012. View at: Publisher Site | Google Scholar
  206. S. Banu, B. Bhaskar, and P. Balasekar, “Hepatoprotective and antioxidant activity of Leucas aspera against D-galactosamine induced liver damage in rats,” Pharmaceutical Biology, vol. 50, no. 12, pp. 1592–1595, 2012. View at: Publisher Site | Google Scholar
  207. R. Rajamurugan, A. Suyavaran, N. Selvaganabathy et al., “Brassica nigra plays a remedy role in hepatic and renal damage,” Pharmaceutical Biology, vol. 50, no. 12, pp. 1488–1497, 2012. View at: Publisher Site | Google Scholar
  208. Z.-Q. Jiang, Y. Li, L.-H. Jiang, H. Gu, and M.-Y. Wang, “Hepatoprotective effects of extracts from processed corni fructus against D-galactose-induced liver injury in mice,” Zhong Yao Cai, vol. 36, no. 1, pp. 85–89, 2013. View at: Google Scholar
  209. S. Zhang, D. Wang, X. Wang et al., “Aqueous extract of Bai-Hu-Tang, a classical Chinese herb formula, prevents excessive immune response and liver injury induced by LPS in rabbits,” Journal of Ethnopharmacology, vol. 149, no. 1, pp. 321–327, 2013. View at: Publisher Site | Google Scholar
  210. X. Li, C. Gou, H. Yang, J. Qiu, T. Gu, and T. Wen, “Echinacoside ameliorates D-galactosamine plus lipopolysaccharide-induced acute liver injury in mice via inhibition of apoptosis and inflammation,” Scandinavian Journal of Gastroenterology, vol. 49, no. 8, pp. 993–1000, 2014. View at: Publisher Site | Google Scholar
  211. J.-L. Zhang, H. Zeng, and X.-B. Wang, “Discussion of Chinese syndrome typing in acute hepatic failure model,” Zhongguo Zhong Xi Yi Jie He Za Zhi, vol. 31, no. 5, pp. 659–662, 2011. View at: Google Scholar
  212. Y.-G. Yang, Y.-W. Liu, H.-Y. Hua, and L.-J. Li, “Effect of Sanhuangyinchi decoction on liver damage and caspase-3 in rats with acute hepatic failure,” Nan Fang Yi Ke Da Xue Xue Bao, vol. 30, no. 11, pp. 2443–2445, 2010. View at: Google Scholar
  213. T. Zhou, X.-C. Yan, Q. Chen et al., “Effects of Chinese herbal medicine Fuzheng Huayu recipe and its components against hepatocyte apoptosis in mice with hepatic injury,” Zhong Xi Yi Jie He Xue Bao, vol. 9, no. 1, pp. 57–63, 2011. View at: Publisher Site | Google Scholar
  214. W. Ma, Y. Yang, J. Diao, Y. Liu, H. Hua, and X. Wen, “Sanhuangyinchi decoction pretreatment ameliorates acute hepatic failure in rats by suppressing antioxidant stress and caspase-3 expression,” Nan Fang Yi Ke Da Xue Xue Bao, vol. 34, no. 4, pp. 482–486, 2014. View at: Google Scholar
  215. F.-L. Wang, H.-Z. Yang, Y.-M. Li, W.-K. Wu, and Z.-C. Zou, “Prevention and treatment mechanism of qingxia therapy (based on yinchenhao decoction and dachengqi decoction) on hepatocyte apoptosis in rats with acute hepatic injury induced by lipopolysaccharide/D-galactosamine,” Zhong Yao Cai, vol. 37, no. 5, pp. 848–852, 2014. View at: Google Scholar
  216. R. Nagalekshmi, A. Menon, D. K. Chandrasekharan, and C. K. K. Nair, “Hepatoprotective activity of Andrographis paniculata and Swertia chirayita,” Food and Chemical Toxicology, vol. 49, no. 12, pp. 3367–3373, 2011. View at: Publisher Site | Google Scholar
  217. D. S. Kushwah, M. T. Salman, P. Singh, V. K. Verma, and A. Ahmad, “Protective effects of ethanolic extract of Nigella sativa seed in paracetamol induced acute hepatotoxicity In vivo,” Pakistan Journal of Biological Sciences, vol. 17, no. 4, pp. 517–522, 2014. View at: Publisher Site | Google Scholar
  218. V. Madhavan, A. S. Pandey, A. Murali, and S. N. Yoganarasimhan, “Protective effects of Capparis sepiaria root extracts against acetaminophen-induced hepatotoxicity in Wistar rats,” Journal of Complementary & Integrative Medicine, vol. 9, article 1, 2012. View at: Google Scholar
  219. N. Anusuya, K. Raju, and S. Manian, “Hepatoprotective and toxicological assessment of an ethnomedicinal plant Euphorbia fusiformis Buch.-Ham.ex D.Don,” Journal of Ethnopharmacology, vol. 127, no. 2, pp. 463–467, 2010. View at: Publisher Site | Google Scholar
  220. A. J. S. J. Samuel, S. Mohan, D. K. Chellappan, A. Kalusalingam, and S. Ariamuthu, “Hibiscus vitifolius (Linn.) root extracts shows potent protective action against anti-tubercular drug induced hepatotoxicity,” Journal of Ethnopharmacology, vol. 141, no. 1, pp. 396–402, 2012. View at: Publisher Site | Google Scholar
  221. M. A. Gbadegesin and O. A. Odunola, “Aqueous and ethanolic leaf extracts of Ocimum basilicum (sweet basil) protect against sodium arsenite-induced hepatotoxicity in Wistar rats,” Nigerian Journal of Physiological Sciences, vol. 25, no. 1, pp. 29–36, 2010. View at: Google Scholar
  222. H. G. Kim, J. S. Lee, J. S. Lee, J. M. Han, and C. G. Son, “Hepatoprotective and antioxidant effects of Myelophil on restraint stress-induced liver injury in BALB/c mice,” Journal of Ethnopharmacology, vol. 142, no. 1, pp. 113–120, 2012. View at: Publisher Site | Google Scholar
  223. Y. S. Al-Awthan, S. M. Hezabr, A. M. Al-Zubairi, and F. A. Al-Hemiri, “Effects of aqueous extract of Withania somnifera on some liver biochemical and histopathological parameters in male guinea pigs,” Pakistan Journal of Biological Sciences, vol. 17, no. 4, pp. 504–510, 2014. View at: Publisher Site | Google Scholar
  224. S. Matić, S. Stanić, D. Bogojević et al., “Methanol extract from the stem of Cotinus coggygria Scop., and its major bioactive phytochemical constituent myricetin modulate pyrogallol-induced DNA damage and liver injury,” Mutation Research/Genetic Toxicology and Environmental Mutagenesis, vol. 755, no. 2, pp. 81–89, 2013. View at: Publisher Site | Google Scholar
  225. S. Ravikumar, M. Gnanadesigan, S. Jacob Inbaneson, and A. Kalaiarasi, “Hepatoprotective and antioxidant properties of Suaeda maritima (L.) Dumort ethanolic extract on concanavalin-A induced hepatotoxicity in rats,” Indian Journal of Experimental Biology, vol. 49, no. 6, pp. 455–460, 2011. View at: Google Scholar
  226. K. Vijayavel, C. Anbuselvam, and B. Ashokkumar, “Protective effect of Coleus aromaticus Benth (Lamiaceae) against naphthalene-induced hepatotoxicity,” Biomedical and Environmental Sciences, vol. 26, no. 4, pp. 295–302, 2013. View at: Publisher Site | Google Scholar
  227. T. Wu, M.-J. Chang, Y.-J. Xu, X.-P. Li, G. Du, and D. Liu, “Protective effect of Calculus bovis sativus on intrahepatic cholestasis in rats induced by alpha-naphthylisothiocyanate,” The American Journal of Chinese Medicine, vol. 41, no. 6, pp. 1393–1405, 2013. View at: Publisher Site | Google Scholar
  228. L.-L. Ding, B.-F. Zhang, W. Dou, L. Yang, C.-S. Zhan, and Z.-T. Wang, “Protective effect of Danning tablet on acute livery injury with cholestasis induced by α-naphthylisothiocyanate in rats,” Journal of Ethnopharmacology, vol. 140, no. 2, pp. 222–229, 2012. View at: Publisher Site | Google Scholar

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