Evidence-Based Complementary and Alternative Medicine http://www.hindawi.com The latest articles from Hindawi Publishing Corporation © 2015 , Hindawi Publishing Corporation . All rights reserved. Validation of a RP-HPLC-DAD Method for Chamomile (Matricaria recutita) Preparations and Assessment of the Marker, Apigenin-7-glucoside, Safety and Anti-Inflammatory Effect Thu, 03 Sep 2015 11:36:50 +0000 http://www.hindawi.com/journals/ecam/2015/828437/ Chamomile is a medicinal plant, which presents several biological effects, especially the anti-inflammatory effect. One of the compounds related to this effect is apigenin, a flavonoid that is mostly found in its glycosylated form, apigenin-7-glucoside (APG), in natural sources. However, the affectivity and safety of this glycoside have not been well explored for topical application. In this context, the aim of this work was to develop and validate a reversed-phase high-performance liquid chromatography (RP-HPLC-DAD) method to quantify APG in chamomile preparations. Additionally, the safety and the anti-inflammatory potential of this flavonoid were verified. The RP-HPLC-DAD method was developed and validated with linearity at 24.0–36.0 μg/mL range (). Intra- and interday precision (RSD) were 0.27–2.66% and accuracy was 98.27–101.21%. The validated method was applied in the analysis of chamomile flower heads, glycolic extract, and Kamillen cream, supporting the method application in the quality control of chamomile preparations. Furthermore, the APG safety was assessed by MTT cytotoxicity assay and mutagenic protocols and the anti-inflammatory activity was confirmed by a diminished TNF- production showed by mice macrophages treated with APG following LPS treatment. Felipe Galeti Miguel, Amanda Henriques Cavalheiro, Nathália Favaretto Spinola, Diego Luis Ribeiro, Gustavo Rafael Mazzaron Barcelos, Lusânia Maria Greggi Antunes, Juliana Issa Hori, Franciane Marquele-Oliveira, Bruno Alves Rocha, and Andresa Aparecida Berretta Copyright © 2015 Felipe Galeti Miguel et al. All rights reserved. Self-Care Practices for Common Colds by Primary Care Patients: Study Protocol of a European Multicenter Survey—The COCO Study Wed, 02 Sep 2015 06:37:47 +0000 http://www.hindawi.com/journals/ecam/2015/272189/ Background. Self-care for common colds is frequent, yet little is known about the spectrum, regional differences, and potential risks of self-care practices in patients from various European regions. Methods/Design. We describe the study protocol for a cross-sectional survey in 27 primary care centers from 14 European countries. At all sites, 120 consecutive adult patients, who visit their general practitioner for any reason, filled in a self-administered 27-item questionnaire. This addresses patients’ self-care practices for common colds. Separately, the subjective level of discomfort when having a common cold, knowing about the diseases’ self-limited nature, and medical and sociodemographic data are requested. Additionally, physicians are surveyed on their use of and recommendations for self-care practices. We are interested in investigating which self-care practices for common colds are used, whether the number of self-care practices used is influenced by knowledge about the self-limited nature of the disease, and the subjective level of discomfort when having a cold and to identify potential adverse interactions with chronic physician-prescribed medications. Further factors that will be considered are, for example, demographic characteristics, chronic conditions, and sources of information for self-care practices. All descriptive and analytical statistics will be performed on the pooled dataset and stratified by country and site. Discussion. To our knowledge, COCO is the first European survey on the use of self-care practices for common colds. The study will provide new insight into patients’ and general practitioners’ self-care measures for common colds across Europe. Birgitta M. Weltermann, Biljana Gerasimovska-Kitanovska, Anika Thielmann, Juliette Chambe, Heidrun Lingner, Enzo Pirrotta, Krzysztof Buczkowski, Selda Tekiner, Slawomir Czachowski, Tamer Edirne, Andrzej Zielinski, Hülya Yikilkan, Tuomas Koskela, Ferdinando Petrazzuoli, Robert D. Hoffman, Marija Petek Šter, Clara Guede Fernández, Ayşegül Uludağ, Kathryn Hoffmann, Vildan Mevsim, and Sanda Kreitmayer Pestic Copyright © 2015 Birgitta M. Weltermann et al. All rights reserved. A Novel Pharmacological Method to Study the Chinese Medicinal Formula Hua-Zheng-Hui-Sheng-Dan Tue, 01 Sep 2015 13:15:51 +0000 http://www.hindawi.com/journals/ecam/2015/436807/ Objectives. Hua-Zheng-Hui-Sheng-Dan (HZHSD) was used as an experimental model to explore research methods of large formulae in traditional Chinese medicine (TCM) using current molecular biology approaches. Materials and Methods. The trypan blue exclusion assay was used to determine cell viability and cell numbers. Flow cytometry was used to assess cell cycle distribution and apoptosis. The concentration of cyclin D1 was analyzed by enzyme-linked immunosorbent assay. The median effect principle was used in drug combination studies. An orthogonal experimental design was used to estimate the effects of each herb at different concentrations. The HeLa xenograft mouse model was used to compare the antitumor activity of drugs in vivo. Results. Among the 35 herbs that comprise HZHSD, Radix Rehmanniae Preparata (RRP), Caesalpinia sappan (CS), Evodia rutaecarpa (ER), Folium Artemisiae Argyi (FAA), Leonurus japonicus Houtt (LJH), Tumeric (Tu), Radix Paeoniae Alba (RPA), and Trogopterus Dung (TD) effectively inhibited the proliferation of HeLa and SKOV3 cells. Only RRR had an effect on HeLa and SKOV3 cell viability. According to the median effect principle, Angelica sinensis (Oliv.) (AS), Tabanus (Ta), and Pollen Typhae (PT), which were proven to have a significant synergistic inhibitory effect on the proliferation of HeLa cells, were added to the original eight positive herbs. The combination of RPA and AS had a synergistic effect on inducing cell cycle S phase arrest and decreasing intracellular cyclin D1 in HeLa cells. By orthogonal experimental design, LJH and Tu were considered unnecessary herbs. The small formula (SHZHSD) consisted of RPA, AS, RRR, Ta., TD, PT, ER, CS, and FAA and was able to inhibit cell proliferation and induce cell apoptosis. The antitumor effects of HZHSD and SHZHSD were also compared in vivo. Conclusions. Through molecular biology approaches both in vitro and in vivo, research into single drugs, and analysis using the median effect principle and orthogonal experimental design, the small formula (SHZHSD) was determined from the original formula (HZHSD). SHZHSD exhibited superior antitumor activity compared with the original formula both in vitro and in vivo. Rui Cao, Hong Zhang, Jie Guo, Xiao-hui Liu, Chang Liu, Cui-hong Zhu, and Xiong-zhi Wu Copyright © 2015 Rui Cao et al. All rights reserved. Effect of Lepidium meyenii Walp. on Semen Parameters and Serum Hormone Levels in Healthy Adult Men: A Double-Blind, Randomized, Placebo-Controlled Pilot Study Tue, 01 Sep 2015 09:28:56 +0000 http://www.hindawi.com/journals/ecam/2015/324369/ Background/Aims. Products of Lepidium meyenii Walp. (maca) are touted worldwide as an alimentary supplement to enhance fertility and restore hormonal balance. Enhancing properties of maca on semen parameters in animals were previously reported by various authors, but we present to the best of our knowledge the first double-blind, randomized, placebo-controlled pilot trial in men. The aim of this study was to evaluate the effects of maca on semen parameters and serum hormone levels in healthy adult men. Methods. A group of 20 volunteers aged 20–40 years was supplied by milled hypocotyl of maca or placebo (1.75 g/day) for 12 weeks. Negative controls of semen were compared to the samples after 6 and 12 weeks of maca administration; negative blood controls were compared to the samples after 12 weeks of treatment. Results. Sperm concentration and motility showed rising trends compared to placebo even though levels of hormones did not change significantly after 12 weeks of trial. Conclusion. Our results indicate that maca possesses fertility enhancing properties in men. As long as men prefer to use alimentary supplement to enhance fertility rather than prescribed medication or any medical intervention, it is worth continuing to assess its possible benefits. Ingrid Melnikovova, Tomas Fait, Michaela Kolarova, Eloy C. Fernandez, and Luigi Milella Copyright © 2015 Ingrid Melnikovova et al. All rights reserved. Overcome Cancer Cell Drug Resistance Using Natural Products Tue, 01 Sep 2015 06:21:22 +0000 http://www.hindawi.com/journals/ecam/2015/767136/ Chemotherapy is one of the major treatment methods for cancer. However, failure in chemotherapy is not uncommon, mainly due to dose-limiting toxicity associated with drug resistance. Management of drug resistance is important towards successful chemotherapy. There are many reports in the Chinese literature that natural products can overcome cancer cell drug resistance, which deserve sharing with scientific and industrial communities. We summarized the reports into four categories: (1) in vitro studies using cell line models; (2) serum pharmacology; (3) in vivo studies using animal models; and (4) clinical studies. Fourteen single compounds were reported to have antidrug resistance activity for the first time. In vitro, compounds were able to overcome drug resistance at nontoxic or subtoxic concentrations, in a dose-dependent manner, by inhibiting drug transporters, cell detoxification capacity, or cell apoptosis sensitivity. Studies in vivo showed that single compounds, herbal extract, and formulas had potent antidrug resistance activities. Importantly, many single compounds, herbal extracts, and formulas have been used clinically to treat various diseases including cancer. The review provides comprehensive data on use of natural compounds to overcome cancer cell drug resistance in China, which may facilitate the therapeutic development of natural products for clinical management of cancer drug resistance. Pu Wang, Hua Li Yang, Ying Juan Yang, Lan Wang, and Shao Chin Lee Copyright © 2015 Pu Wang et al. All rights reserved. Sea Buckthorn Leaf Extract Protects Jejunum and Bone Marrow of 60Cobalt-Gamma-Irradiated Mice by Regulating Apoptosis and Tissue Regeneration Tue, 01 Sep 2015 06:19:50 +0000 http://www.hindawi.com/journals/ecam/2015/765705/ A single dose (30 mg/kg body weight) of standardized sea buckthorn leaf extract (SBL-1), administered 30 min before whole body 60Co-gamma-irradiation (lethal dose, 10 Gy), protected >90% of mice population. The purpose of this study was to investigate the mechanism of action of SBL-1 on jejunum and bone marrow, quantify key bioactive compounds, and analyze chemical composition of SBL-1. Study with 9-week-old inbred male Swiss albino Strain ‘A’ mice demonstrated that SBL-1 treatment before 60Co-gamma-irradiation (10 Gy) significantly () countered radiation induced decreases in jejunum crypts (1.27-fold), villi number (1.41-fold), villus height (1.25-fold), villus cellularity (2.27-fold), cryptal Paneth cells (1.89-fold), and Bcl2 level (1.54-fold). It countered radiation induced increases in cryptal apoptotic cells (1.64-fold) and Bax levels (1.88-fold). It also countered radiation (2 Gy and 3 Gy) induced bone marrow apoptosis (1.59-fold and 1.85-fold) and micronuclei frequency (1.72-fold and 2.6-fold). SBL-1 rendered radiation protection by promoting cryptal stem cells proliferation, by regulating apoptosis, and by countering radiation induced chromosomal damage. Quercetin, Ellagic acid, Gallic acid, high contents polyphenols, tannins, and thiols detected in SBL-1 may have contributed to radiation protection by neutralization of radiation induced oxidative species, supporting stem cell proliferation and tissue regeneration. Madhu Bala, Manish Gupta, Manu Saini, M. Z. Abdin, and Jagdish Prasad Copyright © 2015 Madhu Bala et al. All rights reserved. Topical Hypericum perforatum Improves Tissue Regeneration in Full-Thickness Excisional Wounds in Diabetic Rat Model Mon, 31 Aug 2015 13:47:59 +0000 http://www.hindawi.com/journals/ecam/2015/245328/ Delayed wound healing process is one of the most important concerns in diabetes. Healing of wounds has four phases, namely, hemostasis, inflammation, proliferation, and remodeling. For a successful repair, all four factors must occur properly. Hence, we aimed to evaluate the healing effects of Hypericum perforatum (HP) on full-thickness diabetic skin wounds by using stereological methods. Forty-eight female diabetic rats were randomly divided into four groups (): gel base treated group, HP 5% gel treated group, HP 10% gel treated group, and the control group which received no treatment. A circular 1 cm2 full-thickness wound was created on the animal’s neck and wound area was measured every three days. After sacrificing the animals, skin samples were fixed and prepared for stereological evaluations. Based on the results, HP treated group showed faster wound closure rate in comparison with control and vehicle groups (). In addition, numerical density of fibroblasts, volume density of collagen bundles, and mean diameter and volume densities of the vessels in HP group were significantly higher than control and vehicle groups. The results of this study showed that HP has the ability to improve tissue regeneration by enhancing fibroblast proliferation, collagen bundle synthesis, and revascularization. Soheila Yadollah-Damavandi, Mehdi Chavoshi-Nejad, Ehsan Jangholi, Noushin Nekouyian, Sahar Hosseini, Amin Seifaee, Shima Rafiee, Hossein Karimi, Soheil Ashkani-Esfahani, Yekta Parsa, and Maryam Mohsenikia Copyright © 2015 Soheila Yadollah-Damavandi et al. All rights reserved. Design and Formulation of Optimized Microemulsions for Dermal Delivery of Resveratrol Mon, 31 Aug 2015 13:05:32 +0000 http://www.hindawi.com/journals/ecam/2015/540916/ The objective of this study was to formulate optimal formulations of microemulsions (MEs) and evaluate their feasibility for delivery of resveratrol into human skin ex vivo. Oil-in-water MEs were formulated using surfactant (S) PEG-8 caprylic/capric glycerides and cosurfactant (CoS) polyglyceryl-6-isostearate. Ethyl oleate was used as an oily phase. MEs were formulated using 5 : 1, 6 : 1, and 7 : 1 surfactant and cosurfactant (S : CoS) weight ratios. Pseudoternary phase diagrams were constructed and optimal compositions of MEs were obtained using Design Expert software. Mean droplet size for optimized ME formulations was determined to be 68.54 ± 1.18 nm, 66.08 ± 0.16 nm, and 66.66 ± 0.56 nm for systems with S : CoS weight ratios 5 : 1, 6 : 1, and 7 : 1, respectively. Resveratrol loading resulted in mean droplet size increase. The distribution of droplet size between fractions changed during storage of formulated MEs. Results demonstrated the increase of number of droplets and relative surface area when S : CoS weight ratios were 6 : 1 and 7 : 1 and the decrease when S : CoS weight ratio was 5 : 1. The highest penetration of resveratrol into the skin ex vivo was determined from ME with S : CoS weight ratio 5 : 1. It was demonstrated that all MEs were similar in their ability to deliver resveratrol into the skin ex vivo. Vaida Juškaitė, Kristina Ramanauskienė, and Vitalis Briedis Copyright © 2015 Vaida Juškaitė et al. All rights reserved. Effect of Tongxinluo on Nephrin Expression via Inhibition of Notch1/Snail Pathway in Diabetic Rats Mon, 31 Aug 2015 12:54:32 +0000 http://www.hindawi.com/journals/ecam/2015/424193/ Podocyte injury is an important mechanism of diabetic nephropathy (DN). Accumulating evidence suggests that nephrin expression is decreased in podocyte in DN. Moreover, it has been demonstrated that tongxinluo (TXL) can ameliorate renal structure disruption and dysfunction in DN. However, the effect of TXL on podocyte injury in DN and its molecular mechanism is unclear. In order to explore the effect of TXL on podocyte injury and its molecular mechanism in DN, our in vivo and in vitro studies were performed. Our results showed that TXL increased nephrin expression in diabetic rats and in high glucose cultured podocyte. Meanwhile, TXL decreased ICN1 (the intracellular domain of notch), HES1, and snail expression in podocyte in vivo and in vitro. More importantly, we found that TXL protected podocyte from injury in DN. The results demonstrated that TXL inhibited the activation of notch1/snail pathway and increased nephrin expression, which may be a mechanism of protecting effect on podocyte injury in DN. Fangqiang Cui, Dawei Zou, Yanbin Gao, Na Zhang, Jinyang Wang, Liping Xu, Jianguo Geng, Jiaoyang Li, Shengnan Zhou, and Xinyao Wang Copyright © 2015 Fangqiang Cui et al. All rights reserved. Acupoint Catgut Embedding for Obesity: Systematic Review and Meta-Analysis Mon, 31 Aug 2015 11:39:04 +0000 http://www.hindawi.com/journals/ecam/2015/401914/ Acupoint catgut embedding (ACE) was applied widely to antiweight in China. The aim of this review is to estimate the effectiveness and safety of ACE on obesity. A literature search was conducted in PubMed, Cochrane Library, EBASE, CNKI, and so forth, using combination subject terms of obesity (or overweight, weight loss, etc.) and acupoint catgut embedding (or catgut implantation, catgut embedding). Improvement rate, reduction of body weight and body mass index (BMI), and so forth were analyzed. 43 studies were included for systematic review and meta-analysis. Although with poor methodological quality, ACE was superior to manual acupuncture (MA), sham, and cupping in improvement rate and presented a better tendency () compared with drugs and electroacupuncture (EA). Mean values of weight loss by ACE were 1.14 kg, 1.26 kg, 1.79 kg, and 3.01 kg comparing with MA, drugs, EA, and sham, respectively. Mean of BMI reduced to 0.56 kg/m2, 0.83 kg/m2, 0.79 kg/m2, and 1.63 kg/m2 comparing with MA, drugs, EA, and sham. Less adverse effects were reported. Pooled outcomes presented a tendency of equal or superior effects to other interventions and fewer side effects. Future high quality trials with rigorous design and positive FDA approved drug as control are urgent to assess the effect of ACE for obesity. PROSPERO registration number is as follows: CRD42015016006. Taipin Guo, Yulan Ren, Jun Kou, Jing Shi, Sun Tianxiao, and Fanrong Liang Copyright © 2015 Taipin Guo et al. All rights reserved. Acupuncture Affects Autonomic and Endocrine but Not Behavioural Responses Induced by Startle in Horses Sun, 30 Aug 2015 14:30:35 +0000 http://www.hindawi.com/journals/ecam/2015/219579/ Startle is a fast response elicited by sudden acoustic, tactile, or visual stimuli in a variety of animals and in humans. As the magnitude of startle response can be modulated by external and internal variables, it can be a useful tool to study reaction to stress. Our study evaluated whether acupuncture can change cardiac autonomic modulation (heart rate variability); and behavioural (reactivity) and endocrine (cortisol levels) parameters in response to startle. Brazilian Sport horses were subjected to a model of startle in which an umbrella was abruptly opened near the horse. Before startle, the horses were subjected to a 20-minute session of acupuncture in acupoints GV1, HT7, GV20, and BL52 (ACUP) and in nonpoints (NP) or left undisturbed (CTL). For analysis of the heart rate variability, ultrashort-term (64 s) heart rate series were interpolated (4 Hz) and divided into 256-point segments and the spectra integrated into low (LF; 0.01–0.07 Hz; index of sympathetic modulation) and high (HF; 0.07–0.50 Hz; index of parasympathetic modulation) frequency bands. Acupuncture (ACUP) changed the sympathovagal balance with a shift towards parasympathetic modulation, reducing the prompt startle-induced increase in LF/HF and reducing cortisol levels 30 min after startle. However, acupuncture elicited no changes in behavioural parameters. Julia Dias Villas-Boas, Daniel Penteado Martins Dias, Pablo Ignacio Trigo, Norma Aparecida dos Santos Almeida, Fernando Queiroz de Almeida, and Magda Alves de Medeiros Copyright © 2015 Julia Dias Villas-Boas et al. All rights reserved. Toxic Markers of Matrine Determined Using 1H-NMR-Based Metabolomics in Cultured Cells In Vitro and Rats In Vivo Sun, 30 Aug 2015 12:41:40 +0000 http://www.hindawi.com/journals/ecam/2015/598412/ Matrine is one of the main bioactive alkaloids of Sophora flavescens Aiton, which has been widely used to treat various diseases in China. These diseases include viral hepatitis, liver fibrosis, cardiac arrhythmia, skin diseases, and tumors. However, matrine is also the main toxic compound of this herb, and the available biomarkers are not reliable in detecting or quantifying matrine risk. Metabolomics is a powerful tool used to identify early toxicity biomarkers that are specific indicators of damage to biosystems. This study aimed to find the potential biomarkers of the matrine-induced toxic effects in rats and HepG2 cells. The toxicological effects of rats induced by matrine could be derived from the elevated taurine and trimethylamine N-oxide levels and the depletion in hippurate and tricarboxylic acid cycle intermediates, such as 2-oxoglutarate, citrate, and succinate in the urine. Cell metabolomics revealed that the levels of alanine, choline, glutathione, lactate, phosphocholine, and cholesterol showed dose-dependent decreases, whereas the levels of taurine, fatty acid, and unsaturated fatty acid showed dose-dependent increases. Overall, a significant perturbation of metabolites in response to high dose of matrine was observed both in vivo and in vitro, and the selected metabolites particularly represent an attractive marker for matrine-induced toxicity. Zhonghuang Li, Liang Zheng, Jian Shi, Guiyu Zhang, Linlin Lu, Lijun Zhu, Jiajie Zhang, and Zhongqiu Liu Copyright © 2015 Zhonghuang Li et al. All rights reserved. Ginkgolide C Suppresses Adipogenesis in 3T3-L1 Adipocytes via the AMPK Signaling Pathway Sun, 30 Aug 2015 11:21:54 +0000 http://www.hindawi.com/journals/ecam/2015/298635/ Ginkgolide C, isolated from Ginkgo biloba leaves, is a flavone reported to have multiple biological functions, from decreased platelet aggregation to ameliorating Alzheimer disease. The study aim was to evaluate the antiadipogenic effect of ginkgolide C in 3T3-L1 adipocytes. Ginkgolide C was used to treat differentiated 3T3-L1 cells. Cell supernatant was collected to assay glycerol release, and cells were lysed to measure protein and gene expression related to adipogenesis and lipolysis by western blot and real-time PCR, respectively. Ginkgolide C significantly suppressed lipid accumulation in differentiated adipocytes. It also decreased adipogenesis-related transcription factor expression, including peroxisome proliferator-activated receptor and CCAAT/enhancer-binding protein. Furthermore, ginkgolide C enhanced adipose triglyceride lipase and hormone-sensitive lipase production for lipolysis and increased phosphorylation of AMP-activated protein kinase (AMPK), resulting in decreased activity of acetyl-CoA carboxylase for fatty acid synthesis. In coculture with an AMPK inhibitor (compound C), ginkgolide C also improved activation of sirtuin 1 and phosphorylation of AMPK in differentiated 3T3-L1 cells. The results suggest that ginkgolide C is an effective flavone for increasing lipolysis and inhibiting adipogenesis in adipocytes through the activated AMPK pathway. Chian-Jiun Liou, Xuan-Yu Lai, Ya-Ling Chen, Chia-Ling Wang, Ciao-Han Wei, and Wen-Chung Huang Copyright © 2015 Chian-Jiun Liou et al. All rights reserved. Mechanisms, Efficacy, and Safety of Bacopa monnieri (Brahmi) for Cognitive and Brain Enhancement Sun, 30 Aug 2015 09:05:56 +0000 http://www.hindawi.com/journals/ecam/2015/717605/ Con Stough, Hemant Singh, and Andrea Zangara Copyright © 2015 Con Stough et al. All rights reserved. The Anti-Inflammatory Effect of Prunus yedoensis Bark Extract on Adipose Tissue in Diet-Induced Obese Mice Sun, 30 Aug 2015 06:36:40 +0000 http://www.hindawi.com/journals/ecam/2015/937904/ Chronic, low-grade inflammatory responses occur in obese adipose tissue and play a crucial role in the development of insulin resistance. Macrophages exposed to high glucose upregulate the expression of SRA, a macrophage-specific scavenger receptor. The present study investigated whether Prunus yedoensis (PY) bark extract affects the inflammatory response and scavenger receptor gene expression observed in a diet-induced obesity model in vivo. Oral administration of PY extract significantly reduced fasting blood glucose levels without a change in body weight in mice fed a high fat diet for 17 weeks. PY extract significantly suppressed expression of inflammatory and macrophage genes such as tumor necrosis factor-α, interleukin-6, and F4/80 in epididymal adipose tissue. Among scavenger receptor genes, SRA expression was significantly reduced. The inhibitory responses of PY extract and its fractions were determined through evaluation of scavenger receptor expression in THP-1 cells. PY extract and its ethyl acetate fraction decreased the levels of SRA mRNA and phospho-ERK1/2 during monocyte differentiation. Our data indicate that the anti-inflammatory effects of PY extract and its downregulation of SRA seem to account for its hypoglycemic effects. Hee Kang, Tae-Kyung Kwak, Bo-Geun Kim, and Kyung-Jin Lee Copyright © 2015 Hee Kang et al. All rights reserved. Bacopa monniera (CDRI-08) Upregulates the Expression of Neuronal and Glial Plasticity Markers in the Brain of Scopolamine Induced Amnesic Mice Thu, 27 Aug 2015 13:28:03 +0000 http://www.hindawi.com/journals/ecam/2015/837012/ Preclinical studies on animal models have discerned the antiamnesic and memory-enhancing potential of Bacopa monniera (Brahmi) crude extract and standardized extracts. These studies primarily focus on behavioral consequences. However, lack of information on molecular underpinnings has limited the clinical trials of the potent herb in human subjects. In recent years, researchers highlight plasticity markers as molecular correlates of amnesia and being crucial to design therapeutic targets. In the present report, we have investigated the effect of a special extract of B. monniera (CDRI-08) on the expression of key neuronal (BDNF and Arc) and glial (GFAP) plasticity markers in the cerebrum of scopolamine induced amnesic mice. Pre- and postadministration of CDRI-08 ameliorated amnesic effect of scopolamine by decreasing acetyl cholinesterase activity and drastically upregulating the mRNA and protein expression of BDNF, Arc, and GFAP in mouse cerebrum. Interestingly, the plant extract per se elevated BDNF and Arc expression as compared to control but GFAP was unaltered. In conclusion, our findings provide the first molecular evidence for antiamnesic potential of CDRI-08 via enhancement of both neuronal and glial plasticity markers. Further investigations on detailed molecular pathways would encourage therapeutic application of the extract in memory disorders. Arpita Konar, Akash Gautam, and M. K. Thakur Copyright © 2015 Arpita Konar et al. All rights reserved. Molecular and Functional Characterization of Bacopa monniera: A Retrospective Review Thu, 27 Aug 2015 12:22:49 +0000 http://www.hindawi.com/journals/ecam/2015/945217/ Over the last 50 years, laboratories around the world analyzed the pharmacological effect of Bacopa monniera extract in different dimensions, especially as a nerve tonic and memory enhancer. Studies in animal model evidenced that Bacopa treatment can attenuate dementia and enhances memory. Further, they demonstrate that Bacopa primarily either acts via antioxidant mechanism (i.e., neuroprotection) or alters different neurotransmitters (serotonin (5-hydroxytryptamine, 5-HT), dopamine (DA), acetylcholine (ACh), γ-aminobutyric acid (GABA)) to execute the pharmacological effect. Among them, 5-HT has been shown to fine tune the neural plasticity, which is a substrate for memory formation. This review focuses on the studies which trace the effect of Bacopa treatment on serotonergic system and 5-HT mediated key molecular changes that are associated with memory formation. Koilmani Emmanuvel Rajan, Jayakumar Preethi, and Hemant K. Singh Copyright © 2015 Koilmani Emmanuvel Rajan et al. All rights reserved. Sauropus androgynus (L.) Merr. Induced Bronchiolitis Obliterans: From Botanical Studies to Toxicology Thu, 27 Aug 2015 12:13:24 +0000 http://www.hindawi.com/journals/ecam/2015/714158/ Sauropus androgynus L. Merr. is one of the most popular herbs in South Asia, Southeast Asia, and China where it was known as a slimming agent until two outbreaks of pulmonary dysfunction were reported in Taiwan and Japan in 1995 and 2005, respectively. Several studies described that the excessive consumption of Sauropus androgynus could cause drowsiness, constipation, and bronchiolitis obliterans and may lead to respiratory failure. Interestingly, this herb has been used in Malaysia and Indonesia in cooking and is commonly called the “multigreen” or “multivitamin” plant due to its high nutritive value and inexpensive source of dietary protein. The plant is widely used in traditional medicine for wound healing, inducing lactation, relief of urinary disorders, as an antidiabetic cure and also fever reduction. Besides these medicinal uses, the plant can also be used as colouring agent in food. This review will explore and compile the fragmented knowledge available on the botany, ethnobotany, chemical constitutes, pharmacological properties, and toxicological aspects of this plant. This comprehensive review will give readers the fundamental, comprehensive, and current knowledge regarding Sauropus androgynus L. Merr. Hamidun Bunawan, Siti Noraini Bunawan, Syarul Nataqain Baharum, and Normah Mohd. Noor Copyright © 2015 Hamidun Bunawan et al. All rights reserved. Purification and Characterization of Asparaginase from Phaseolus vulgaris Seeds Thu, 27 Aug 2015 11:51:32 +0000 http://www.hindawi.com/journals/ecam/2015/309214/ L-asparaginase from bacteria has been used in treatment of acute lymphoblastic leukemia. The aim of this study was to purify and characterize L-asparaginase from Phaseolus vulgaris seeds instead of microbial sources. L-asparaginase was purified to apparent homogeneity. The enzyme has molecular mass of 79 kDa. The purified asparaginase had very low activity toward a number of asparagine and glutamine analogues. L-asparaginase was free from glutaminase activity. Kinetic parameters, Km and Vmax of purified enzyme, were found to be 6.72 mM and 0.16 μM, respectively. The enzyme had optimum pH at 8.0. The enzyme showed high stability at alkaline pH (pH 7.5–9.0) when incubated for up to 24 h. L-asparaginase had the same temperature optimum and thermal stability at 37°C. K+ was able to greatly enhance the activity of asparaginase by 150% compared with other metals tested. In conclusion, L-asparaginase showed no glutaminase activity and good stability over a wide range of physiological conditions, and thus it could be used as a potential candidate for treatment of acute lymphoblastic leukemia. Saleh A. Mohamed, Mohamed F. Elshal, Taha A. Kumosani, and Alia M. Aldahlawi Copyright © 2015 Saleh A. Mohamed et al. All rights reserved. A Special Extract of Bacopa monnieri (CDRI-08) Restores Learning and Memory by Upregulating Expression of the NMDA Receptor Subunit GluN2B in the Brain of Scopolamine-Induced Amnesic Mice Thu, 27 Aug 2015 11:32:35 +0000 http://www.hindawi.com/journals/ecam/2015/254303/ In the present communication, we have investigated effects of the CDRI-08, a well characterized extract of Bacopa monnieri, on expression of the GluN2B subunit of NMDAR in various brain regions of the scopolamine-induced amnesic mice. Our behavioral data reveal that scopolamine-treated amnesic mice exhibit significant decline in the spatial memory compared to the normal control mice. Our RT-PCR and immunoblotting data revealed that the scopolamine treatment resulted in a significant downregulation of the NMDAR GluN2B subunit expression in prefrontal cortex and hippocampus. Our enzyme assay data revealed that scopolamine caused a significant increase in the acetylcholinesterase activity in both the brain regions. Further, oral administration of the CDRI-08 to scopolamine-treated amnesic mice restored the spatial memory which was found to be associated with significant upregulation of the GluN2B subunit expression and decline in the acetylcholinesterase activity in prefrontal cortex as well as hippocampus towards their levels in the normal control mice. Our study provides the evidence for the mechanism underlying role of the Bacopa monnieri extract (CDRI-08) in restoring spatial memory in amnesic mice, which may have therapeutic implications. Rakesh Rai, Hemant K. Singh, and S. Prasad Copyright © 2015 Rakesh Rai et al. All rights reserved. CDRI-08 Attenuates REST/NRSF-Mediated Expression of NMDAR1 Gene in PBDE-209-Exposed Mice Brain Thu, 27 Aug 2015 11:27:31 +0000 http://www.hindawi.com/journals/ecam/2015/403840/ CDRI-08 is a standardized bacoside enriched ethanolic extract of Bacopa monnieri, a nootropic plant. We reported that CDRI-08 attenuated oxidative stress and memory impairment in mice, induced by a flame retardant, PBDE-209. In order to explore the mechanism, present study was designed to examine the role of CDRI-08 on the expression of NMDAR1 (NR1) and the binding of REST/NRSF to NR1 promoter against postnatal exposure of PBDE-209. Male mice pups were orally supplemented with CDRI-08 at the doses of 40, 80, or 120 mg/kg along with PBDE-209 (20 mg/kg) during PND 3–10 and frontal cortex and hippocampus were collected at PND 11 and 60 to study the expression and regulation of NR1 by RT-PCR and electrophoretic mobility shift assay, respectively. The findings showed upregulated expression of NR1 and decreased binding of REST/NRSF to NR1 promoter after postnatal exposure of PBDE-209. Interestingly, supplementation with CDRI-08 significantly restored the expression of NR1 and binding of REST/NRSF to NR1 promoter near to the control value at the dose of 120 mg/kg. In conclusion, the results suggest that CDRI-08 possibly acts on glutamatergic system through expression and regulation of NR1 and may restore memory, impaired by PBDE-209 as reported in our previous study. Priya Verma, Rajaneesh K. Gupta, Behrose S. Gandhi, and Poonam Singh Copyright © 2015 Priya Verma et al. All rights reserved. Bacopa monnieri Extract (CDRI-08) Modulates the NMDA Receptor Subunits and nNOS-Apoptosis Axis in Cerebellum of Hepatic Encephalopathy Rats Thu, 27 Aug 2015 11:26:13 +0000 http://www.hindawi.com/journals/ecam/2015/535013/ Hepatic encephalopathy (HE), characterized by impaired cerebellar functions during chronic liver failure (CLF), involves N-methyl-D-aspartate receptor (NMDAR) overactivation in the brain cells. Bacopa monnieri (BM) extract is a known neuroprotectant. The present paper evaluates whether BM extract is able to modulate the two NMDAR subunits (NR2A and NR2B) and its downstream mediators in cerebellum of rats with chronic liver failure (CLF), induced by administration of 50 mg/kg bw thioacetamide (TAA) i.p. for 14 days, and in the TAA group rats orally treated with 200 mg/kg bw BM extract from days 8 to 14. NR2A is known to impart neuroprotection and that of NR2B induces neuronal death during NMDAR activation. Neuronal nitric oxide synthase- (nNOS-) apoptosis pathway is known to mediate NMDAR led excitotoxicity. The level of NR2A was found to be significantly reduced with a concomitant increase of NR2B in cerebellum of the CLF rats. This was consistent with significantly enhanced nNOS expression, nitric oxide level, and reduced Bcl2/Bax ratio. Moreover, treatment with BM extract reversed the NR2A/NR2B ratio and also normalized the levels of nNOS-apoptotic factors in cerebellum of those rats. The findings suggest modulation of NR2A and NR2B expression by BM extract to prevent neurochemical alterations associated with HE. Papia Mondal and Surendra Kumar Trigun Copyright © 2015 Papia Mondal and Surendra Kumar Trigun. All rights reserved. Bacopa monnieri as an Antioxidant Therapy to Reduce Oxidative Stress in the Aging Brain Thu, 27 Aug 2015 11:23:42 +0000 http://www.hindawi.com/journals/ecam/2015/615384/ The detrimental effect of neuronal cell death due to oxidative stress and mitochondrial dysfunction has been implicated in age-related cognitive decline and neurodegenerative disorders such as Alzheimer’s disease. The Indian herb Bacopa monnieri is a dietary antioxidant, with animal and in vitro studies indicating several modes of action that may protect the brain against oxidative damage. In parallel, several studies using the CDRI08 extract have shown that extracts of Bacopa monnieri improve cognitive function in humans. The biological mechanisms of this cognitive enhancement are unknown. In this review we discuss the animal studies and in vivo evidence for Bacopa monnieri as a potential therapeutic antioxidant to reduce oxidative stress and improve cognitive function. We suggest that future studies incorporate neuroimaging particularly magnetic resonance spectroscopy into their randomized controlled trials to better understand whether changes in antioxidant status in vivo cause improvements in cognitive function. Tamara Simpson, Matthew Pase, and Con Stough Copyright © 2015 Tamara Simpson et al. All rights reserved. Validation of Quantitative HPLC Method for Bacosides in KeenMind Thu, 27 Aug 2015 11:23:23 +0000 http://www.hindawi.com/journals/ecam/2015/696172/ Brahmi (Bacopa monnieri) has been used by Ayurvedic medical practitioners in India for almost 3000 years. The pharmacological properties of Bacopa monnieri were studied extensively and the activities were attributed mainly due to the presence of characteristic saponins called “bacosides.” Bacosides are complex mixture of structurally closely related compounds, glycosides of either jujubogenin or pseudojujubogenin. The popularity of herbal medicines and increasing clinical evidence to support associated health claims require standardisation of the phytochemical actives contained in these products. However, unlike allopathic medicines which typically contain a single active compound, herbal medicines are typically complex mixtures of various phytochemicals. The assay for bacosides in the British Pharmacopoeia monograph for Bacopa monnieri exemplifies that only a subset of bacosides present are included in the calculation of total bacosides. These results in calculated bacoside values are significantly lower than those attained for the same material using more inclusive techniques such as UV spectroscopy. This study illustrates some of the problems encountered when applying chemical analysis for standardisation of herbal medicines, particularly in relation to the new method development and validation of bacosides from KeenMind. Ashley Dowell, George Davidson, and Dilip Ghosh Copyright © 2015 Ashley Dowell et al. All rights reserved. Bacoside A: Role in Cigarette Smoking Induced Changes in Brain Thu, 27 Aug 2015 11:21:28 +0000 http://www.hindawi.com/journals/ecam/2015/286137/ Cigarette smoking (CS) is a major health hazard that exerts diverse physiologic and biochemical effects mediated by the components present and generated during smoking. Recent experimental studies have shown predisposition to several biological consequences from both active and passive cigarette smoke exposure. In particular, passive smoking is linked to a number of adverse health effects which are equally harmful as active smoking. A pragmatic approach should be considered for designing a pharmacological intervention to combat the adverse effects of passive smoking. This review describes the results from a controlled experimental condition, testing the effect of bacoside A (BA) on the causal role of passive/secondhand smoke exposure that caused pathological and neurological changes in rat brain. Chronic exposure to cigarette smoke induced significant changes in rat brain histologically and at the neurotransmitter level, lipid peroxidation states, mitochondrial functions, membrane alterations, and apoptotic damage in rat brain. Bacoside A is a neuroactive agent isolated from Bacopa monnieri. As a neuroactive agent, BA was effective in combating these changes. Future research should examine the effects of BA at molecular level and assess its functional effects on neurobiological and behavioral processes associated with passive smoke. G. Vani, K. Anbarasi, and C. S. Shyamaladevi Copyright © 2015 G. Vani et al. All rights reserved. A Special Extract of Bacopa monnieri (CDRI-08)-Restored Memory in CoCl2-Hypoxia Mimetic Mice Is Associated with Upregulation of Fmr-1 Gene Expression in Hippocampus Thu, 27 Aug 2015 11:13:16 +0000 http://www.hindawi.com/journals/ecam/2015/347978/ Fragile X mental retardation protein (FMRP) is a neuronal translational repressor and has been implicated in learning, memory, and cognition. However, the role of Bacopa monnieri extract (CDRI-08) in enhancing cognitive abilities in hypoxia-induced memory impairment via Fmr-1 gene expression is not known. Here, we have studied effects of CDRI-08 on the expression of Fmr-1 gene in the hippocampus of well validated cobalt chloride (CoCl2)-induced hypoxia mimetic mice and analyzed the data with alterations in spatial memory. Results obtained from Morris water maze test suggest that CoCl2 treatment causes severe loss of spatial memory and CDRI-08 is capable of reversing it towards that in the normal control mice. Our semiquantitative RT-PCR, Western blot, and immunofluorescence microscopic data reveal that CoCl2-induced hypoxia significantly upregulates the expression of Hif-1α and downregulates the Fmr-1 expression in the hippocampus, respectively. Further, CDRI-08 administration reverses the memory loss and this is correlated with significant downregulation of Hif-1α and upregulation of Fmr-1 expression. Our data are novel and may provide mechanisms of hypoxia-induced impairments in the spatial memory and action of CDRI-08 in the recovery of hypoxia led memory impairment involving Fmr-1 gene encoded protein called FMRP. Anupama Rani and S. Prasad Copyright © 2015 Anupama Rani and S. Prasad. All rights reserved. The Stimulative Effect of Yangjing Capsule on Testosterone Synthesis through Nur77 Pathway in Leydig Cells Thu, 27 Aug 2015 09:16:18 +0000 http://www.hindawi.com/journals/ecam/2015/408686/ Yangjing Capsule (YC), an innovative Chinese medicine based on traditional prescription, promotes testosterone synthesis by upregulating the expression of steroidogenic enzymes. Nur77 as a nuclear receptor is known to regulate the expression of many steroid synthetases. This study aimed to explore the potential mechanisms by which YC regulates testosterone synthesis in Leydig cells. Real-time PCR and Western blot analysis were employed to assess the expressions of steroidogenic enzymes and Nur77 after treating MLTC-1 cells with YC. The luciferase reporter gene assay was performed to detect the activity of Nur77 gene promoter. Also, the expressions of steroid synthases were detected after Nur77 gene was knocked down. YC significantly stimulated Nur77 production and upregulated StAR and HSD3B expression, and this agrees with the activity of Nur77 gene promoter that was significantly enhanced by YC. Interestingly, knockdown of Nur77 blocked the above YC’s effects and consequently inhibited testosterone synthesis in MLTC-1 cells. YC promotes StAR and HSD3B expression and upregulates testosterone synthesis in Leydig cells, which is mediated by Nur77 pathway. Yalong Gu, Xindong Zhang, Dalin Sun, Hongle Zhao, Bin Cai, Chao Gao, Li Gao, Yugui Cui, Zhian Tang, and Baofang Jin Copyright © 2015 Yalong Gu et al. All rights reserved. Bioactives and Traditional Herbal Medicine for the Treatment of Cardiovascular/Cerebrovascular Diseases 2015 Wed, 26 Aug 2015 11:54:20 +0000 http://www.hindawi.com/journals/ecam/2015/320545/ Joen-Rong Sheu, Pitchairaj Geraldine, and Mao-Hsiung Yen Copyright © 2015 Joen-Rong Sheu et al. All rights reserved. Effects of Tetramethylpyrazine on Functional Recovery and Neuronal Dendritic Plasticity after Experimental Stroke Wed, 26 Aug 2015 08:08:38 +0000 http://www.hindawi.com/journals/ecam/2015/394926/ The 2,3,5,6-tetramethylpyrazine (TMP) has been widely used in the treatment of ischemic stroke by Chinese doctors. Here, we report the effects of TMP on functional recovery and dendritic plasticity after ischemic stroke. A classical model of middle cerebral artery occlusion (MCAO) was established in this study. The rats were assigned into 3 groups: sham group (sham operated rats treated with saline), model group (MCAO rats treated with saline) and TMP group (MCAO rats treated with 20 mg/kg/d TMP). The neurological function test of animals was evaluated using the modified neurological severity score (mNSS) at 3 d, 7 d, and 14 d after MCAO. Animals were euthanized for immunohistochemical labeling to measure MAP-2 levels in the peri-infarct area. Golgi-Cox staining was performed to test effect of TMP on dendritic plasticity at 14 d after MCAO. TMP significantly improved neurological function at 7 d and 14 d after ischemia, increased MAP-2 level at 14 d after ischemia, and enhanced spine density of basilar dendrites. TMP failed to affect the spine density of apical dendrites and the total dendritic length. Data analyses indicate that there was significant negative correlation between mNSS and plasticity measured at 14 d after MCAO. Thus, enhanced dendritic plasticity contributes to TMP-elicited functional recovery after ischemic stroke. Jun-Bin Lin, Chan-Juan Zheng, Xuan Zhang, Juan Chen, Wei-Jing Liao, and Qi Wan Copyright © 2015 Jun-Bin Lin et al. All rights reserved. Biothermodynamic Assay of Coptis-Evodia Herb Couples Wed, 26 Aug 2015 07:20:10 +0000 http://www.hindawi.com/journals/ecam/2015/565364/ Objective. To illustrate the difference in cold/hot natural properties and therapeutic effect of coptis-evodia herb couples by using cold/hot plate differentiating technology and microcalorimetry combined with material basis analysis in vivo and in vitro. It showed that animal retention ratio in hot pad significantly decreased along with the decrease in coptis proportion in coptis-evodia herb couples. In addition, Zuojin wan markedly reduced the retention ratio of gastritis mice in the hot pad, while Fanzuojin wan displayed an opposite result. Further, Mg2+-ATPase, Ca2+-ATPase, and T-AOC activity significantly weakened in coptis-treated group in the livers of the mice. In the gastric cells from the gastritis mice, Fanzuojin wan remarkably increased calorific value for growth and metabolism, while Zuojin wan significantly reduced the calorigenic effect. It suggested that the changes in the major chemical compositions (especially alkaloids) were the material base-induced transformation between “cold” and “hot” syndromes. The material basis which affected the transformation between “cold” and “hot” syndromes might be X2, X3, X4, X8, epiberberine hydrochloride, jatrorrhizine hydrochloride, coptisine sulphate, palmatine hydrochloride, and berberine hydrochloride. The CHPD combined with microcalorimetry technology is a good method to determine the differences in the “cold” and “hot” natural properties of coptis-evodia herb couples. Hongbo Yang, Min Su, Qi Yao, Yanling Zhao, Danhong Chen, and Lei Jia Copyright © 2015 Hongbo Yang et al. All rights reserved.