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

Table 1

Animal models of NAFLD and NASH used to investigate the effect of herbal medicines.

Model (ingredients)	Duration	Herbal medicine	Pathological and biochemical changes; mechanisms involved	Reference
Diet induced models (high calories/fats diet)
10% lard oil and 2% cholesterol	4 wk	Si 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 tablets	4 wk	Alkaloids of Rubus alceifolius	Reduced 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-80	56 d	Schisandra chinensis Baill	Reduced SOD, serum TC, and LDLC and increased hepatic MDA.	[85]
12% lard oil, 2% cholesterol, 0.2% propylthiouracil, and 0.5% bile salt	6 wk	Sapindus 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 wk	TZQ 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% cholesterol	4 wk + 4 wk treatment	Jiang Zhi Granule	Improved hepatic steatosis, reduced ALT and AST, and improved free fatty acid and triacylglycerol levels. Reduced LXR alpha and SREBP-1c.	[88]
MCD diet	8 wk	Fuzheng Huayu recipe	Model 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 diet	8 wk	Ping-tang recipe	Suppressed 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 diet	9 wk	Puerariae radix, Lycium barbarum, Crataegus pinnatifida, and Polygonati rhizoma	Reduced fasting blood glucose and improved insulin resistance.	[89]
10% lard, 1.5% cholesterol, 0.2% sodium deoxycholate, 5% sugar, 0.05% prothiopyrimidine	10 wk (5 wk treatment)	Yiqi huoxue decoction and solutions of herbs	Hepatic 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 wk	Lycium barbarum polysaccharides	Reduced 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 diet	12 wk	Chinese medicine recipes	Treatment group blocked fatty degeneration. Decreased expressions of JNK and p-JNK.	[91]
5.3 kcal/g (fat 59%, protein 16%, and carbohydrate 24%)	15 wk	Yin-Chen-Hao-Tang	Reduced 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% carbohydrate	16 wk	Garcinia cambogia supplement	Increased 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 wk	Chaihu-Shugan-San and Shen-ling-bai-zhu-San	Model 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 diet	16 wk	Soothing liver and invigorating spleen recipes	Reduced TC, LDL-C, TC, and TG in treatment group. Reduced TLR4 expression.	[95]
21.9 kJ/g, 60% fat, 20% protein, and 20% carbohydrate	20 wk	Troxerutin	Decreased 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/g	24 wk	Crude extract of Lycium barbarum polysaccharide	Lowered 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 : 1	4 mo + 40 d treatment	Rhein	Steatotic 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% cholesterol	12 wk	Salvia-Nelumbinis naturalis	Reduced macrovesicular steatosis, TG, LDL-C, and FFA levels. Increased IRS and Akt phosphorylation and decreased SOCS3.	[97]
High fat diet		Cigu Xiaozhi pills	Model 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 diet	4 wk	Huanglian Jiedu decoction	Ameliorated pathological changes in fatty liver. Associated with increase of M2 macrophages populations.	[20]
MCD diet in db/db mice	4 wk	Sho-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 wk	pomegranate flowers polyphenols	Reduced fat drops, non-HDL-C, and transaminase. Antioxidant ability enhanced and PON1 increased in liver.	[19]
High fat diet (10% lard and 2% cholesterol) + CCl4	8 wk	Dangyao (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) + CCl4	8 wk	Dangfei Liganning capsules	Improved steatosis, hepatocyte ballooning, and inflammatory infiltration as observed in model group. Improved MDA and ALT.	[99]
10% lard + 2% cholesterol + CCl4	8 wk	Soothing liver and invigorating spleen recipes	Reduced SREBP-1 and SCD-1.	[100]
High cholesterol diet in KK-Ay mice	10 wk	Corosolic acid	Reduced blood cholesterol and liver cholesterol content. Inhibited activity of cholesterol acyltransferase.	[101]
High fat diet (640 kcal/100 g) + gold-thioglucose	12 wk	Bofutsushosan	Blocked 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 wk	Goshajinkigan	Reduced AST. Increased body and adipose tissue weight and reduced elevated liver weight.	[102]

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