Ethnopharmacology

Protective role of wild garlic on isoproterenol-induced myocardial necrosis in wistar rats Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Saravanakumar Murugesan, Arunagiri Pandiyan, Lakshmi Saravanakumar, Kogi Moodley, Irene Mackraj Abstract Ethnopharmacological relevance Tulbaghia violacea Harv. (TVL) is a folk medicine, native to South Africa which has previously shown antioxidant, anti-hypertensive and anti-diabetic effects. The Aim of the study: The aim of the current study was to investigate the protective role of wild garlic or TVL on isoproterenol (ISO)-induced myocardial necrosis in rats. Materials and methods Animal (n = 6 each group) were pre and co-treated with TVL (60 mg/kg body weight) daily for 30 days. Myocardial necrosis was administrated by subcutaneous injection of ISO (85 mg/kg body weight) into rats on 29th and 30th day. On the 31st day, rats were anaesthetized and blood, heart samples were obtained for the biochemical, histopathological and molecular study. The specific protein target analysis from TVL was done by reverse docking study (reverse pharmacophore mapping) using PharmMapper. Results The levels of cardiac markers, lipid peroxidation products, and heart rate were considerably increased in ISO-induced myocardial necrosis in rats whilst plasma enzymatic antioxidants were significantly decreased. Myocardial necrotic mRNA genes were increased in ISO-induced myocardial necrosis in rats compared to controls. Pre and co-treatment with TVL and ramipril of myocardial necrosis in rats showed significant effects on all the biochemical and molecular studies evaluated. TVL reduced heart rate, prevented oxidative stress and downregulated the Fas-receptor and caspase-mediated apoptosis-signaling pathway, and heart muscle damage in myocardial necrosis in rats. The specific target protein [disulfide, bis (2-sulfhydrylethyl] from TVL mediates the protective effects. Conclusion Wild garlic or TVL extract has shown a protective effect on ISO-induced myocardial necrosis in rats by increasing antioxidant production confirmed with docking studies. Graphical abstract

Effect of saffron (stigma of Crocus sativus L.) aqueous extract on ethanol toxicity in rats: A biochemical, histopathological and molecular study Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Alireza Rezaee-Khorasany, Bibi Marjan Razavi, Elahe Taghiabadi, Abbas Tabatabaei Yazdi, Hossein Hosseinzadeh Abstract Ethnopharmacological relevance Saffron (Crocus sativus L.) is considered in the Iranian traditional medicine because of many therapeutic properties such as sedative agent, strengthen the stomach and liver, improving the uterus disorders and infectious wounds. The detoxification of alcohol was one of the most important of saffron effects in ancient medicine. Aim of the study In the current research, the protective effects of saffron aqueous extract (Aq. Ext.) versus oxidative stress, apoptosis, inflammation, histopathological and biochemical abnormalities induced by ethanol were evaluated. Materials & methods The male Wistar rats were divided into seven groups consisted of 6 rats in control (distilled water), ethanol (5 g/kg - 50% v/v), Aq. Ext. (40, 80 and 160 mg/kg) plus ethanol, Aq. Ext. 80 and 160 mg/kg. Animals were treated for four weeks and at the end of treatment period, histopathological damages, biochemical markers, apoptosis, levels of MDA and GSH, TNF-α and IL-6 were evaluated. Results Ethanol induced nephrotoxicity and hepatotoxicity as evidenced by histopathological damages and biochemical abnormalities. The level of MDA was significantly enhanced while GSH content was remarkably reduced in ethanol-treated rats, but protective groups restored them. Also, the levels of TNF-α and IL-6 were regulated by Aq. Ext. Furthermore, the effects of ethanol on histopathological and biochemical parameters were improved by Aq. Ext. The ethanol increased the expression of Bax/Bcl2 ratio, caspase-3, -8, and -9. Real-time PCR and western blot analysis proved that Aq. Ext. treatment inhibited apoptosis induced by ethanol through decreasing the Bax/Bcl2 ratio (mRNA and protein) and caspases-3, -8, and -9 levels in the kidney and liver. Conclusion The results of this research demonstrated that Aq. Ext. could exert protective effects against ethanol toxicity in rat kidney and liver via antioxidant, anti-apoptosis, and anti-inflammatory effects. Graphical abstract

In vivo anti-inflammatory activity and UPLC-MS/MS profiling of the peels and pulps of Cucumis melo var. cantalupensis and Cucumis melo var. reticulatus Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Shahira M. Ezzat, Mai Raslan, Maha M. Salama, Esther T. Menze, Seham S. El Hawary Abstract Ethnopharmacological relevance Cucumis melo var. cantalupensis and Cucumis melo var. reticulatus are the most famous varieties of netted muskmelon or cantaloupe in Egypt. Cantaloupe has a great reputation as an anti-inflammatory drug for hot inflammation of liver, cough, eczema, and kidney disorders such as ulcers in the urinary tract, and our objective was to confirm this use scientifically. Materials and methods Inflammation was induced in adult male Sprague Dawley rats by subcutaneous injection of 0.05 ml of carrageenan (1% solution in saline) into the plantar surface of the right hind paw 30 min after oral pretreatment of the rats with 95% ethanolic extracts of Cucumis melo var. cantalupensis peels (CCP) and pulps (CCU) and Cucumis melo var. reticulatus peels (CRP) and pulps (CRU) at doses of 25 and 50 mg/kg. Indomethacin (10 mg/kg) was used as a standard drug. The effect of the tested samples was measured on the oedema volume, as well as PGE-2, TNF-α, IL-6 and IL-1β levels. Metabolic profiling of the extracts was performed using UPLC-MS/MS analysis. Results Pretreatment of rats with the ethanol extract of the pulps and peels of the two varieties at doses of 25 and 50 mg/kg significantly inhibited the carrageenan-induced increase in the oedema volume of the rat paws after 3 h, except for the low dose of the French cantaloupe pulp. CRP at 50 mg/kg caused the most significant reductions in both TNF-α (P < 0.05) and IL-1β (P < 0.001) levels, while CCP caused the most significant reductions in PGE-2 and IL-6 (P < 0.05) levels. Increases in PGE-2, TNF-α, IL-6 and IL-1β levels were also significantly prevented by indomethacin (10 mg/kg). UPLC-MS/MS facilitated the identification of 44 phenolic compounds, including phenolic acids and flavonoids. Conclusion This is the first report of the chemical and biological study of the peels of Cucumis melo var. cantalupensis and Cucumis melo var. reticulatus. Graphical abstract

Bodies of the plant and Animal Kingdom: An illustrated manuscript on materia medica in the Netherlands (ca. 1800) Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Ingeborg Swart, Mieke Beumer, Wouter Klein, Tinde van Andel Abstract Ethnopharmacological relevance Around 1800, Amsterdam was a global trade hub for materia medica of Dutch, European and exotic origin. Contemporary knowledge on medicinal plants in academic circles has been well documented in local pharmacopoeia, illustrated herbals and catalogues of botanic gardens. Until the end of the ancient regime, physicians, surgeons and apothecaries were trained how to use plants in their specific guild or Collegium Medicum. Little is known, however, on how the plant collectors and merchants that provided the pharmaceutical substances to apothecaries learnt to recognise the variety of medicinal products. Aim of the study To analyse the content, origin, purpose and scientific importance of an anonymous, undated, hand-written Dutch manuscript on materia medica, entitled Corpora ex Regno Vegetabili/Animali (Bodies of the Plant/Animal kingdom) kept by the Artis Library of the University of Amsterdam, the Netherlands. Materials and methods We digitised the entire manuscript and dated the paper by means of its watermark. We identified the plant and animal species using the historic Dutch and Latin names, the illustrations and historic literature. We compared the plant properties and uses to contemporary literature to check whether the information in the manuscript was original or copied from another source. Results The paper was produced between 1759 and 1816 in Zaandam, the Netherlands. The manuscript contains 19 substances of animal origin, one mineral and 273 plants and plant-derived products, which belong to ca. 260 species. While most plants are native or cultivated in the Netherlands, 111 plant entries (105 spp.) represent exotic products, imported from as far as Madagascar and Australia. A total of 134 illustrations were cut out from a 1549 Dutch edition of the New Herbal by Leonhard Fuchs (1543), but only 69% correspond to the correct species. The manuscript contains detailed descriptions on growth locations, field characteristics, flowering season, provenance and quality of the medicinal products, including methods to detect forgery. The author mostly described humoral properties of the plants rather than listing medicinal recipes. We did not find evidence that he copied his texts from other sources, but the Dutch and Latin names correspond largely with the Amsterdam pharmacopoeia from 1795. Conclusions The author's extensive knowledge on trade names, quality and origin of materia medica and his refrain from using literature suggests he could have been a merchant, an intermediary between herb cultivators, overseas traders and apothecaries. This manuscript offers a unique insight in the global trade in medicinal products and the circulation of knowledge in non-academic circles around 1800. Graphical abstract

The anti-diabetic activities, gut microbiota composition, the anti-inflammatory effects of Scutellaria–coptis herb couple against insulin resistance-model of diabetes involving the toll-like receptor 4 signaling pathway Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Chang-hua Zhang, Jun-qing Sheng, Surendra Sarsaiya, Fu-xing Shu, Tong-tong Liu, Xiu-ying Tu, Guang-qiang Ma, Guo-Liang Xu, Hong-xiang Zheng, Li-fen Zhou Abstract Ethnopharmacological relevance Scutellaria–coptis herb couple (SC) is one of the well-known herb couples in many traditional Chinese compound formulas used for the treatment of diabetes mellitus (DM), which has been used to treat DM for thousands of years in China. Aim of the study Few studies have confirmed in detail the anti-diabetic activities of SC in vivo and in vitro. The present investigations aimed to evaluate the anti-diabetic activity of SC in type 2 diabetic KK-Ay mice and in RAW264.7 macrophages to understand its possible mechanism. Materials and methods High-performance liquid chromatography with ultraviolet detection (HPLC-UV) and LC-LTQ-Orbitrap Pro mass spectrometry were used to analyze the active ingredients of SC extracts and control the quality. A type 2 diabetic KK-Ay mice model was established by high-fat diet. Body weight, fasting blood glucose levels, fasting blood insulin levels, glycosylated hemoglobin and glycosylated serum protein were measured. The effects of SC on total cholesterol (TC), high-density lipoprotein (HDL) and triglyceride (TG) levels were examined. The lipopolysaccharide (LPS), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-α) levels were measured. Gut microbial communities were assayed by polymerase chain reaction (PCR) and PCR-denaturing gradient gel electrophoresis (PCR-DGGE) methods. The expressions of Toll-like receptor 4 (TLR4) and MyD88 protein in the colons were measured by western blot. In RAW264.7 macrophages, IL-6, TNF-α, TLR4 and MyD88 protein levels were measured by enzyme-linked immunosorbent assay (ELISA) kits or western blot, and the mRNA expression of IL-6, TNF-α and TLR4 was examined by the real time PCR. Results The present results showed that the SC significantly increased blood HDL and significantly reduced fasting blood glucose, fasting blood insulin, glycosylated hemoglobin, glycosylated serum protein, TC, TG, LPS, IL-6 and TNF-α levels (P < 0.05 or P < 0.01) in type-2 diabetic KK-Ay mice. Furthermore, SC could regulate the structure of intestinal flora. Additionally, the expressions of TLR4 and MyD88 protein in the colons were significantly decreased in the model group (P < 0.05 or P < 0.01). However, SC had no significant effect on weight gain. In RAW264.7 macrophages, SC containing serum (SC-CS) (5%, 10% and 20%) significantly decreased IL-6, TNF-α, TLR4 and MyD88 protein levels and the mRNA expression of IL-6, TNF-α and TLR4 (P < 0.05 or P < 0.01). Conclusions The anti-diabetic effects of SC were attributed to its regulation of intestinal flora and anti-inflammation involving the TLR4 signaling pathway. These findings provide a new insight into the anti-diabetic application for SC in clinical settings and display the potential of SC in the treatment of DM. Graphical abstract

An extract from Myracrodruon urundeuva inhibits matrix mineralization in human osteoblasts Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Adriana Arruda Matos, Flávia Amadeu Oliveira, Alessandra Cury Machado, Luiz Leonardo Saldanha, Cintia Kazuko Tokuhara, Leonardo Perez Souza, Wagner Vilegas, Thiago José Dionísio, Carlos Ferreira dos Santos, Camila Peres-Buzalaf, Anne Lígia Dokkedal, Rodrigo Cardoso de Oliveira Abstract Ethnopharmacological relevance Phytotherapy based on plant-derived compounds is an alternative medicinal strategy for the relief of symptoms and the curing of diseases. The leaves of Myracrodruon urundeuva a medicinal plant also known as "aroeira", has been used in traditional medicine as healing, antiulcer and anti-inflammatory to treat skeletal diseases in Brazil, but its role in bone cell toxicity, as well as in bone formation, remains to be established. Aim of the study We sought to determine the in vitro osteogenic effects of a hydroalcoholic M. urundeuva leaves extract in primary human osteoblasts. Materials and methods Cell viability, reactive oxygen species (ROS) production, alkaline phosphatase (ALP) activity and matrix mineralization were evaluated by MTT assay, DCFH-DA probe, colorimetric-based enzymatic assay and Alizarin Red-staining, respectively. Besides, the matrix metalloproteinase (MMP)-2 and progressive ankylosis protein homolog (ANKH) gene expression were determined by real-time RT-qPCR and MMP-2 activity by zymography. Results Exposure of osteoblasts to M. urundeuva extract significantly decreased viability and increased reactive oxygen species (ROS) production, regardless of the extract concentration. The M. urundeuva extract at 10 μg/mL also downregulated matrix metalloproteinase (MMP)-2, while upregulating progressive ankylosis protein homolog (ANKH) gene expression. By contrast, the MMP-2 activity was unchanged. The M. urundeuva extract at 10 μg/mL also reduced alkaline phosphatase (ALP) activity and mineralization. Conclusions Overall, our findings suggest that the inhibition of osteogenic differentiation and matrix mineralization promoted by M. urundeuva may be due more to an increase in oxidative stress than to the modulation of MMP-2 and ANKH expression. Graphical abstract

Effect of water extract from Berberis heteropoda Schrenk roots on diarrhea-predominant irritable bowel syndrome by adjusting intestinal flora Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Hui-min Zhu, Li Li, Song-ya Li, Qi Yan, Fei Li Abstract Background The aim of the present study was to investigate the antidiarrheal effect of Berberis heteropoda Schrenk roots (BHS), which are used by Chinese minorities to treat diarrhea, through regulation of intestinal flora and related signaling pathways. Methods Wistar rats were randomly divided into 6 groups: Control group (Con), Model group (Mod), three BHS groups (BHS-L (0.65 g/kg), BHS-M (1.955 g/kg), BHS-H (5.86 g/kg) and Bifidobacterium group (Bif). The model of diarrhea-based irritable bowel syndrome (D-IBS) was induced by intragastric administration combined with restraint stress. The CRD method was used to determine the AWR score and the Bristol fecal score. Quantification of the intestinal bacteria groups in feces was performed using colony counting on plates. The mRNA expression levels of Gpr41, Gpr43, TLR2, TLR4, and nuclear protein κB were determined by qRT-PCR, and the relative abundances of intestinal flora in the intestinal contents were determined by high-throughput gene sequencing ratios. Results Oral administration of BHS (L, M and H) significantly reduced the AWR score and the Bristol fecal score, significantly relieved diarrhea in D-IBS rats, reduced the number of Enterococci and Enterobacteria in feces, increased the number of Bifidobacteria and Lactobacilli, and upregulated the expression of SCFA in plasma. qRT-PCR analysis showed that the expression of TLR2, TLR4, Gpr41, Gpr43 and NF-κB in the BHS groups was downregulated. D-IBS rats reduced the abundance and diversity of intestinal flora and BHS (L, M and H) regulated the abundance and diversity of their intestinal flora. Conclusion The above data suggest that BHS potentially alleviates diarrhea, intestinal flora disorder and intestinal inflammation in D-IBS rats by regulating the immunological pathways. BHS is a promising agent in the treatment of D-IBS. Graphical abstract

In vitro genotoxic and antigenotoxic effects of cynarin Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Esra Erikel, Deniz Yuzbasioglu, Fatma Unal Abstract Ethnopharmacological relevance Cynarin is an artichoke phytochemical that possesses a variety of pharmacological features including free-radical scavenging and antioxidant activity. The origin of artichoke species appears to be Mediterranean region. Two of these species, globe artichoke (Cynara cardunculus var. scolymus L.) and cardoon (Cynara cardunculus var. altilis DC), are widely cultivated and consumed. This vegetable, as the basis of the mediterranean diet, has been used as herbal medicine for its therapeutic effects since ancient times. Therefore, this study was performed to determine genotoxic and antigenotoxic effects of cynarin against MMC (mitomycin C) and H2O2(hydrogen peroxide) induced genomic instability using chromosome aberrations (CAs), sister chromatid exchanges (SCEs), micronucleus (MN), and comet assays in human lymphocytes. Materials and methods Lymphocytes obtained from two healthy volunteers (1 male and 1 female) were exposed to different concentrations of cynarin (12–194 μM) alone and the combination of cynarin and MMC (0.60 μM) or cynarin and H2O2(100 μM, only for comet assay). Results Cynarin alone did not induce significant genotoxic effect in the CA, SCE (except 194 μM), MN, and comet assays. The combination of some concentrations of cynarin and MMC decreased the frequency of CAs, SCEs and MN induced by MMC. Furthermore, the combination of cynarin and H2O2 reduced all comet parameters at all the concentrations compared to H2O2 alone. While the highest concentrations of cynarin significantly decreased mitotic index (MI), the combination of cynarin and MMC increased the reduction of MI induced by MMC alone. Conclusion All the results obtained in this study demonstrated that cynarin exhibited antigenotoxic effects rather than genotoxic effects. It is believed that cynarin can act as a potential chemo-preventive against genotoxic agents. Graphical abstract

Raffia palm (Raphia hookeri G. Mann & H. Wendl) wine modulates glucose homeostasis by enhancing insulin secretion and inhibiting redox imbalance in a rat model of diabetes induced by high fructose diet and streptozotocin Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Ochuko L. Erukainure, Olajumoke A. Oyebode, Omamuyovwi M. Ijomone, Chika I. Chukwuma, Neil A. Koorbanally, Md. Shahidul Islam Abstract Ethnopharmacological relevance Raffia palm (Raphia hookeri) wine (RPW) is amongst the natural products from plants, utilized singly or in combination with other medicinal plants for the treatment of several ailments including Diabetes Mellitus (DM). However, there is a scientific dearth on its antidiabetic activity. Aim The antidiabetic effect of RPW and its possible mechanism of actions were investigated in diabetic rats. Methods Four groups of male SD rats were first supplied with 10% fructose solution ad libitum for 2 weeks instead of drinking water followed by an intraperitonial injection of streptozotocin (40 mg/kg) to induce diabetes. Two diabetic groups were administered RPW at 150 and 300 mg/kg bodyweight (BW) respectively; a group was administered with metformin, while the other one was served as a negative control. Two groups of normal rats were administered with water and RPW (300 mg/kg BW) and served as normal control and normal toxicology group, respectively. Results Five weeks treatment of RPW led to significant (p < 0.05) increase in serum insulin and HDL-c levels with concomitant reduction in blood glucose, fructosamine, ALT, uric acid, triglycerides and LDL-c levels in diabetic rats. Rats treated with RPW had elevated levels of GSH, SOD, catalase, ATPase and α-amylase activities, while reduced NO level and myeloperoxidase activity was observed in their serum and pancreatic tissues. RPW also improved pancreatic β-cell function and restored β- and acinar cells morphology, and capillary networks. The activities of glycogen phosphorylase, fructose 1,6 biphosphatase, glucose-6-phosphatase, and acetylcholinesterase were also inhibited in RPW-treated diabetic rats, with concomitant down regulation of Nrf2 gene expression. Conclusion The data of this study suggest that RPW modulates glucose homeostasis by enhancing insulin secretion as well as inhibiting redox imbalance in diabetic rats, which may be attributed to the synergetic effects of its phytochemical constituents as identified by GC-MS analysis. Graphical abstract

Essential oil from Fructus Alpinia zerumbet (fruit of Alpinia zerumbet (Pers.) Burtt.et Smith) protected against aortic endothelial cell injury and inflammation in vitro and in vivo Publication date: 12 June 2019 Source: Journal of Ethnopharmacology, Volume 237 Author(s): Yun-peng Ji, Ting-yu Shi, Yan-yan Zhang, Dan Lin, Ke-gang Linghu, Yi-ni Xu, Ling Tao, Qing Lu, Xiang-chun Shen Abstract Ethnopharmacological relevance Fructus Alpinia zerumbet (FAZ), a dry and ripe fruit of Alpinia zerumbet (Pers.) Burtt. et Smith, is widely used as a spice to treat cardiovascular diseases in clinic as a miao folk medicine in Guizhou Province of China. Essential oil extracted from FAZ (EOFAZ) is the key bioactive ingredients. Aim of the study This study aimed to examine the effects and mechanisms of EOFAZ on lipopolysaccharide (LPS)-induced endothelial cell injury, inflammation and apoptosis in vitro and in vivo. Materials and methods For the in vitro study, LPS-treated human aortic endothelial cells were used to perform PCR, western blot analysis and immunofluorescence. For the in vivo study, male mouse were divided into four groups, vehicle control group and LPS group received 0.5% Tween-80 in saline; and two EOFAZ groups receive different dose of EOFAZ (90 mg kg −1·day−1, 180 mg kg −1·day−1) respectively. Each group was fed for 7 days by intragastrical administration at daily base. Then, except vehicle control group received saline, mice in other three groups were administered with LPS (1 mg kg −1, dissolved in saline) by intraperitoneal injection. 24 h later, Aorta tissue was collected and frozen immediately in liquid N2, stored at −80 °C for western blot analysis. Results We found that EOFAZ completely prevented LPS-induced HAEC activation and inflammation in vitro and in vivo, as assessed by expression of endothelial adhesion molecules, ICAM-1 and VCAM-1. Similarly, EOFAZ significantly blunted LPS-induced endothelial injury, as tested by MTT assay, LDH release and caspase-3 activation. We further demonstrated that TLR4-dependent NF-κB signaling may be involved in the process. Conclusion EOFAZ protected against LPS-induced endothelial cell injury and inflammation likely via inhibition of TLR4-dependent NF-κB signaling. Graphical abstract