Ethnopharmacology

Inhibitory effect of ethanol extract of Ampelopsis brevipedunculata rhizomes on atopic dermatitis-like skin inflammation Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Young-Ae Choi, Ju-Hee Yu, Hong Dae Jung, Soyoung Lee, Pil-Hoon Park, Hyun-Shik Lee, Taeg Kyu Kwon, Tae-Yong Shin, Seung Woong Lee, Mun-Chul Rho, Young Hyun Jang, Sang-Hyun Kim Abstract Ethnopharmacological relevance Extracts from various parts of Ampelopsis brevipedunculata has been used as anti-inflammatory agents in Asian folk medicine. Aim of the study: To demonstrate the medicinal effect of the A. brevipedunculata in skin inflammation, specifically atopic dermatitis (AD). Materials and methods The effect of ethanol extract of A. brevipedunculata rhizomes (ABE) on AD was examined using an AD-like skin inflammation model induced by repeated exposure to house dust mite (Dermatophagoides farinae extract, DFE) and 2,4-dinitrochlorobenzene (DNCB). The mechanism study was performed using tumor necrosis factor (TNF)-α and interferon (IFN)-γ-activated human keratinocytes (HaCaT). Serum histamine and immunoglobulin levels were quantified using enzymatic kits, while the gene expression of cytokines and chemokines was analyzed using quantitative real time polymerase chain reaction. The expression of signaling molecules was detected using Western blot. Results Oral administration of ABE alleviated DFE/DNCB-induced ear thickening and clinical symptoms, as well as immune cell infiltration (mast cells and eosinophils) into the dermal layer. Serum Immunoglobulin (Ig) E, DFE-specific IgE, IgG2a, and histamine levels were decreased after the administration of ABE. ABE also inhibited CD4+IFN-γ+ and CD4+IL-4+ lymphocyte polarization in lymph nodes and expression of TNF-α, IFN-γ, IL-4, IL-13, and IL-31 in the ear tissue. In TNF-α/INF-γ-stimulated keratinocytes, ABE inhibited the gene expression of TNF-α, IL-6, IL-1β, and CCL17. In addition, ABE decreased the nuclear localization of signal transducer and activator of transcription 1 and nuclear factor-κB, and the phosphorylation of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase. Conclusion Collectively, our data demonstrate the pharmacological role and signaling mechanism of ABE in the regulation of skin allergic inflammation, which supports our suggestion that ABE could be developed as a potential therapeutic agent for the treatment of AD. Graphical abstract

Anti-inflammatory activities of Canarium subulatum Guillaumin methanol extract operate by targeting Src and Syk in the NF-κB pathway Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Eunju Choi, Mi-Yeon Kim, Jae Youl Cho Abstract Ethnopharmacological relevance Canarium subulatum Guillaumin is an herbal medicinal plant native to Southeast Asia. Ethnopharmacological evidence suggests that plants of the genus Canarium cure a variety of inflammatory diseases. Aim of the study The pharmacological mechanisms of C. subulatum Guillaumin remain poorly understood. In this study, we investigate inflammatory mechanisms and target molecules using C. subulatum Guillaumin methanol extract (Cs-ME) in inflammatory reactions managed by macrophages. Materials and methods To identify the anti-inflammatory activities of Cs-ME, lipopolysaccharide (LPS)-stimulated macrophages and a murine HCl/EtOH-induced gastritis model were chosen. The luciferase reporter gene assay, Western blot analysis, overexpression strategy, and the cellular thermal shift assay (CETSA) were employed to investigate the molecular mechanisms and target enzymes of Cs-ME. The active ingredients of this extract were also determined by HPLC. Results Released levels of nitric oxide (NO) and mRNA expression levels of iNOS and IL-6 were downregulated by Cs-ME without exhibiting cytotoxicity. This extract inhibited MyD88-induced promoter activity and the nuclear translocation of nuclear factor (NF)-κB. Moreover, we found that Cs-ME reduced the phosphorylation of NF-κB upstream signaling molecules including IκBα, IKKα/β, Src, and Syk in LPS-stimulated macrophage-like RAW264.7 cells. The results of Western blot and CETSA confirmed that Src and Syk are anti-inflammatory targets of Cs-ME. In addition, orally injected Cs-ME alleviated HCl/EtOH-induced gastric ulcers in mice. HPLC analysis indicated that quercetin, luteolin, and kaempferol are major active components of this extract with anti-inflammatory activity. Conclusions Cs-ME exhibits anti-inflammatory effects in vitro and in vivo by targeting Src and Syk in the NF-κB signaling pathway. Consequently, Cs-ME could be developed as an anti-inflammatory herbal medicine. Graphical abstract

Anti-obesity effect of argel (Solenostemma argel) on obese rats fed a high fat diet Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Riham A. El-shiekh, Dalia A. Al-Mahdy, Samar M. Mouneir, Mohamed S. Hifnawy, Essam A. Abdel-Sattar Abstract Ethnopharmacological relevance Solenostemma argel (Argel) is a desert plant commonly used in Egyptian and Sudanese traditional medicine to suppress appetite, for treatment of diabetes, and as an antispasmodic and anti-inflammatory agent. Previously the anti-diabetic, hypolipidemic and lipase inhibitory activities of Argel were reported in animal studies and in-vitro assays. However, its specific mechanism of action as an anti-obesity agent has not been studied before. Aim of the study Assessment of the possible anti-obesity effect of Solenostemma argel on diet-induced obesity and elucidation of its mechanism of action, as well as, standardization of the active plant extract. Materials and methods The ethanolic extract (EtOH-E) and its fractions (CH2Cl2-F: methylene chloride and BuOH-F: n-butanol) were prepared from the aerial parts of S. argel and studied at two dose levels; 200 and 400 mg kg−1 in a model of high fat diet (HFD) fed rats. The animals (72 Male Wister rats) were assigned into 9 groups: group (i) fed with normal diet and groups (ii-iv) fed with high fat diet (HFD) for 16 weeks and treated with orlistat, EtOH-E, CH2Cl2-F and BuOH-F in the beginning of the 8th week. At the end of the experiment, blood samples were analysed for lipid and liver biomarkers, glucose and insulin levels, as well as, adipokines and inflammatory markers. Liver and adipose tissues were examined histopathologically and their homogenates were used to determine levels of oxidative stress markers and lipogenesis-related genes. Body weight was monitored weekly during the experiment. Results Our data showed that consumption of S. argel significantly controlled weight gain, attenuated liver steatosis, improved the lipid profile, modulated adipokines activities, increased β-oxidation gene expression, as well as, decreased the expression of lipogenesis-related genes and ameliorated inflammatory and lipid peroxidation derangement. The ethanolic extract was also standardized using LC–MS analysis for its content of stemmoside C. Conclusions The current study revealed that S. argel is a promising Egyptian natural drug, rich in pregnane glycosides, and could be considered a new therapeutic candidate targeting obesity. Graphical abstract

Evaluation of the schistosomicidal, antioxidant and anti-inflammatory activities of the ethyl acetate fraction from Ozoroa pulcherrima Schweinf. Roots on Schistosoma mansoni-induced liver pathology in mice and its phytochemical characterization Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Hermine Boukeng Jatsa, Nestor Gipwe Feussom, Ulrich Membe Femoe, Mérimé Christian Kenfack, Emilienne Tienga Nkondo, Joseph Bertin Kadji Fassi, Nadège Distele Simo, Cyriaque Moaboulou, Calvine Noumedem Dongmo, Christelle Dongmo Tsague, Etienne Dongo, Pierre Kamtchouing, Louis-Albert Tchuem Tchuente Abstract Ethnopharmacological relevance Ozoroa pulcherrima Schweinf. (syn.: Heeria pulcherrrima Schweinf.) is a small shrub belonging to the family Anacardiaceae. In Africa, the stem and the leaves are used to treat dystocia, hyperthermia, and conjunctivitis, while the root is used to treat dysmenorrhea and intestinal helminthiasis. Aim of the study The aim of this study was to assess the schistosomicidal, antioxidant and anti-inflammatory effects of the ethyl acetate fraction from O. pulcherrima roots methanolic extract (EAOp) on S. mansoni- induced liver pathology in mice. Additionally, its phytochemical composition was elucidated. Material and methods The phytochemical characterization of EAOp was carried out by High-Performance Liquid Chromatography-Mass spectrometry (HPLC-MS). Total phenolic and flavonoid contents were also quantified in the fraction. S. mansoni-infected mice received daily and per os, for 28 days, EAOp at 200 or 400 mg/kg, starting from the 36th day post-infection. Praziquantel was used as reference drug. Uninfected-untreated, uninfected-treated and infected-untreated mice served as controls. At the 65th day post-infection mice were sacrificed and parasitological burden monitored. Transaminases, total bilirubin, and total proteins levels were determined in the plasma. Malondialdehyde (MDA), nitrites, superoxide dismutase (SOD), catalase (CAT) and reduced glutathione (GSH) levels were measured in the liver as biomarkers of the oxidative stress. Liver histology and morphometric analysis of granulomas were also conducted. Results The HPLC-MS analysis data of EAOp revealed the presence of four triterpenes namely oleaterminaloic acid, hydroxyoleanolic acid, moronic acid, and oleanolic acid; a flavonoid dipentoxybenzoic acid and two alkaloids. Its total phenolic content was 76.46 ± 0.01 mg GAE/g and total flavonoid content 6.26 ± 0.31 mg rutin equivalent/g. The reductions of worm burden (48.89 and 75.56%), fecal egg count (77.76 and 69.52%) and egg load in the liver (65.33 and 77.18%) and intestine (78.06 and 84.63%) were significant after EAOp treatment. EAOp at all doses significantly (p < 0.001) reversed the increasing transaminases activities and total bilirubin level induced by the infection. A normalization of total proteins concentration was also recorded. Treatment of S. mansoni-infected mice with EAOp at 200 or 400 mg/kg resulted in a significant reduction (p < 0.001) of MDA concentration by 73.20% and 67.78% respectively. The level of nitrites which was reduced by the infection significantly increased after the treatment. EAOp significantly increased by 4.67 and 5.69-fold the CAT activity and by 126.67% the GSH level. Histologically, a significant reduction of the number (66.39 and 57.82%) and the volume (52.25 and 34.81%) of liver inflammatory granulomas was recorded after EAOp treatment at all doses. Conclusions These results suggest that the liver pathology in S. mansoni infection is improved by EAOp which disclosed good schistosomicidal, antioxidant and anti-inflammatory activities. Its effects on the liver dysfunction and the hepatic oxidative stress were comparable to that of praziquantel. These findings justified the traditional use of O. pulcherrima for the treatment of intestinal helminthiasis. This fraction can be considered as a promising source for schistosomicidal agents. Graphical abstract

Preclinical study of safety of Dendropanax morbifera Leveille leaf extract: General and genetic toxicology Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Jun-Won Yun, Seung-Hyun Kim, Yun-Soon Kim, Eun Jin Choi, Ji-Ran You, Eun-Young Cho, Jung-Hee Yoon, Euna Kwon, Hyoung-Chin Kim, Ja-June Jang, Jin-Sung Park, Jeong-Hwan Che, Byeong-Cheol Kang Abstract Ethnopharmacology relevance Dendropanax morbifera Leveille (DM) has been used in traditional medicines for infectious and skin diseases, and dysmenorrhea. It exhibits a diverse therapeutic potential including anti-cancer, anti-thrombotic, anti-diabetic, anti-oxidant, and anti-inflammatory activities. Aim of the study Despite promising health benefits of DM, knowledge of its potential adverse effects is very limited. The current study focused on the investigation of subchronic toxicity and genotoxicity of extract obtained from DM according to the test guidelines published by the Organization for Economic Cooperation and Development. Materials and methods We conducted a toxicological evaluation of DM extracts using 14-day repeated-dose toxicity study and 13-week repeated-dose toxicity study in Sprague-Dawley rats administered orally at doses of 500, 1000, or 2000 mg/kg/day. The clastogenicity of DM extract was also evaluated by in vitro chromosome aberration assay and in vivo micronucleus assay. Results Assessment of subchronic toxicity of DM extract by oral administration in rats revealed unremarkable treatment-related findings with respect to food/water consumption, body weight, mortality, urinalysis, hematology, serum biochemistry, necropsy, organ weight and histopathology at doses of 500, 1000, and 2000 mg/kg. Accordingly, the level of no-observed-adverse-effect for DM extract in 13-week subchronic toxicity study was considered to be 2000 mg/kg/day in rats. The data observed from in vitro chromosome aberration assay and in vivo micronucleus assay exclude any clastogenicity of DM extract. Conclusion The results suggest that the oral consumption of DM extract has no adverse effects in humans and represents a safe traditional medicine. Graphical abstract

Curcuma longa L. ameliorates asthma control in children and adolescents: A randomized, double-blind, controlled trial Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Gabriel Manarin, Daniela Anderson, Jorgete Maria e Silva, Juliana da Silva Coppede, Persio Roxo-Junior, Ana Maria Soares Pereira, Fabio Carmona Abstract Ethnopharmacological relevance Roots of Curcuma longa L. are used as medicine for millennia. They possess several pharmacological properties, including anti-inflammatory action, and can be suitable for asthma treatment. Aim of the study We aimed to test the hypothesis that, in children and adolescents with persistent asthma, the administration of powdered roots of C. longa for 6 months, in addition to standard treatment, compared to placebo, will result in better disease control. Patients and methods We conducted a randomized, double-blind, placebo-controlled, phase II clinical trial. Patients were randomly assigned to receive 30 mg/kg/day of C. longa for 6 months, or placebo. Data were collected prospectively. All patients were categorized for asthma severity and control according to GINA-2016 and underwent pulmonary function tests. Results Overall, both groups experienced amelioration of their frequency of symptoms and interference with normal activity, but no differences were found between the two treatment groups. However, patients receiving C. longaexperienced less frequent nighttime awakenings, less frequent use of short-acting β-adrenergic agonists, and better disease control after 3 and 6 months. Conclusion The powdered roots of C. longa led to less frequent nighttime awakenings, less frequent use of short-acting β-adrenergic agonists, and better disease control after 3 and 6 months, when compared to placebo. Graphical abstract

Evaluation of the subacute toxicity of Yongdamsagan-tang, a traditional herbal formula, in Crl:CD Sprague Dawley rats Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Eunsook Park, Mee-Young Lee, Chang-Seob Seo, Hyeun-Kyoo Shin, Su-Cheol Han, Hyekyung Ha Abstract Ethnopharmacological relevance Yongdamsagan-tang, a traditional herbal formula, is used widely for the treatment of inflammatory and viral diseases. However, the safety of Yongdamsagan-tang has not been established. Aim of the study To evaluate the subacute toxicity of Yongdamsagan-tang water extract (YSTE) in Crl:CD Sprague Dawley rats. Materials and methods We evaluated the subacute toxicity of YSTE in male and female Crl:CD Sprague Dawley rats (n = 5 per group). Rats were treated with YSTE at doses of 0, 1000, 2000 and 5000 mg/kg administered once a day by oral gavage for 4 weeks. Results There were no significant changes in mortality, body weight, food intake, serum biochemistry, or results of hematology and urinalysis after YSTE administration. However, all rats treated with 5000 mg/kg/day YSTE exhibited excessive salivation and discolored urine. Necropsy findings showed discoloration in the liver of both male (n = 1) and female (n = 3) rats treated with 5000 mg/kg/day YSTE, and an increase in the relative weights of kidney and liver was also found in male rats treated with 5000 mg/kg/day. In addition, decreases in serum creatinine, total bilirubin, alanine transaminase, and alkaline phosphatase were observed in male rats treated with 2000 or 5000 mg/kg/day YSTE. Conclusions Abnormalities in some rats are considered to be independent of YSTE toxicity. Therefore, the results suggest that oral administration of YSTE in rats for 4 weeks is safe at doses of up to 5000 mg/kg/day. Graphical abstract

Huo Xue Tong Luo capsule ameliorates osteonecrosis of femoral head through inhibiting lncRNA-Miat Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Bin Fang, Ying Li, Chen Chen, Qiushi Wei, Jiaqian Zheng, Yamei Liu, Wei He, Dingkun Lin, Gang Li, Yonghui Hou, Liangliang Xu Abstract Ethnopharmacological relevance Traditional Chinese medicine has a long history of treating various bone diseases including osteoporosis and osteonecrosis etc. In clinical treatment, Huo Xue Tong Luo capsule (HXTL capsule) containing Peach kernel, Safflower carthamus, Angelica sinensis, Ligusticum wallichii etc, is one of the mostly used prescriptions for treating osteonecrosis of the femoral head (ONFH) with promising effects. Objectives This study aims to identify the underlying molecular mechanism of how HXTL capsule exerts its function to ameliorate ONFH. Materials and methods All femoral bone tissues were collected during surgeries. Rat bone marrow mesenchymal stem cells (rMSCs) were used. Quantitative real time PCR was used to check the relative expression levels of genes. ChIP assay was performed to evaluate the binding of H3K4me3 and H3K27me3 in Miat promoter. Results We showed that HXTL capsule promoted osteogenesis in rat MSCs as demonstrated by quantitative real time PCR and Alizarin Red S staining. Then we found silencing the endogenous lncRNA-Miat could promote osteogenesis of rMSCs. In addition, the ChIP assay showed that HXTL capsule significantly increased occupancy of H3K27me3 and decreased H3K4me3 in promoter regions of Miat, meaning HXTL capsule inhibited Miat expression through histone modifications. At last, by examining the femoral heads samples obtained from patients with ONFH during total hip arthroplasty surgery, we found the RNA level of hMiat in necrotic tissue was much higher than that of normal tissue. Conclusions Taken together, our study shows that lncRNA-Miat might play an important role in pathogenesis of ONFH, and HXTL capsule can promote osteogenesis to ameliorate ONFH through inhibiting the transcriptional expression of Miat, at least partially. Graphical abstract

Urginea indica attenuated rheumatoid arthritis and inflammatory paw edema in diverse animal models of acute and chronic inflammation Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Ghazala Akhtar, Arham Shabbir Abstract Ethnopharmacological relevance Urginea indica has been used in the traditional system of medicine to treat various inflammatory diseases. Aim of the study Present study investigates the effects of aqueous and ethanolic extracts of U. indica on joint inflammation using different models of acute and chronic inflammation. Materials and methods FCA-induced arthritic rat model, a model of chronic joint inflammation, was used to evaluate the anti-arthritic effects of plant extracts (500 mg/kg, each extract). Macroscopic arthritic scoring, digital water plethysmometery, and histopathological evaluation (H & E staining) were performed to measure the severity of arthritis. Acute inflammatory models like, carrageenan-, histamine- and serotonin-induced paw edema models were used to evaluate effects of U. indica, and supported by xylene-induced ear edema model. Results Both extracts significantly inhibited arthritic development, paw edema, bone erosion, pannus formation, and infiltration of inflammatory cells. Treatment with U. indica extracts resulted in almost normalization of altered counts of white blood cells (WBCs), platelets, and red blood cells (RBCs), along with Hb content. Both extracts were found safe in terms of hepatotoxicity and nephrotoxicity as determined by non-significant difference of alanine aminotransferase (ALT), aspartate transaminase (AST), urea, and creatinine levels among all groups. U. indica significantly attenuated carrageenan-induced paw edema. There are several mechanisms involved in the attenuation of carrageenan-induced paw edema; inhibition of autacoids is one of those important mechanisms. The autacoid inhibition was confirmed by reduction of histamine- and serotonin-induced paw edema found in plant extract treated groups. Suppression of xylene-induced ear edema by plant extract further validated the suggested mechanism of autacoid inhibition. GC-MS analysis showed the presence of isopropyl palmitate in the highest quantity (26.852%). Conclusions This study validated the folkloric uses of U. indica and showed that plant possessed anti-arthritic and anti-inflammatory properties which might be ascribed to inhibition of autacoids. Graphical abstract

Subchronic toxicity and concomitant toxicokinetics of long-term oral administration of total alkaloid extracts from seeds of Peganum harmala Linn: A 28-day study in rats Publication date: 28 June 2019 Source: Journal of Ethnopharmacology, Volume 238 Author(s): Youxu Wang, Hanxue Wang, Liuhong Zhang, Yunpeng Zhang, Yuchen Sheng, Gang Deng, Shuping Li, Ning Cao, Huida Guan, Xuemei Cheng, Changhong Wang Abstract Ethnopharmacological relevance The seeds of Peganum harmala Linn, in which the most abundant active compounds are harmaline and harmine, have been widely used as a traditional medicine in various countries to treat a broad spectrum of diseases including asthma, cough, depression, Parkinson's and Alzheimer's diseases. However, few studies on long-term or subchronic toxicity of seeds of P. harmala were reported after overdose. Aim of the study To investigate the subchronic toxicity and concomitant toxicokinetics of total alkaloid extracts from seeds of P. harmala (TAEP) after oral administration for four weeks in rats. Materials and methods The subchronic toxicity and concomitant toxicokinetics of TAEP were evaluated after 28-day oral administration in rats at daily dose levels of 15, 45, and 150 mg/kg. The signs of toxicity and mortality were monitored and recorded daily. The body weight and average food consumption were measured weekly. The analyses of hematology, biochemistry, urine, relative organ weights and histopathology were conducted at the termination of treatment and recovery phase. For concomitant toxicokinetics study, the plasma toxicokinetic parameters, tissue distribution, and excretion of predominant ingredients harmaline and harmine in TAEP and metabolites harmalol and harmol were tested. Results Following initial repeated exposure to high-dose (150 mg/kg/day) of TAEP excitotoxic reaction, such as tremor, was observed, but tolerated on the fourth day after multiple dosing. The significant alterations in blood glucose and lipid metabolism in liver were observed, but recovered after four weeks of drug withdrawal. The no-observed-adverse-effect level (NOAEL) of TAEP was considered to be 45 mg/kg/day under the present study conditions. There were no significant gender differences in most indexes of subchronic toxicity throughout the experimental period with the exception of food consumption and body weight. In concomitant toxicokinetics study, the alterations of dynamic characteristic for harmaline, harmine and metabolite harmol after multiple oral administration at three doses had been observed. Harmaline, harmine and metabolites harmalol and harmol were widely distributed in organs and there was no accumulation in the tissues examined. The reduction of harmaline and metabolite harmalol in brain after multiple dosing at dose of 150 mg/kg might be closely related to the tremor tolerance. The main excretory pathway for metabolites harmalol and harmol was urinary excretion via kidney. Conclusions The results revealed that TAEP at doses of 15 and 45 mg/kg/day in rats might be safe. Excitotoxic reaction such as tremor occurred initially at dose of 150 mg/kg/day, however, the toxicity was tolerant and reversible. In addition, harmaline and harmine in TAEP had a quick absorption into blood and metabolized to harmalol and harmol, and there was no drug accumulation in the detected tissues. Further studies should be investigated to clarify the mechanisms of tremor tolerance and neurotoxicity of TAEP. Graphical abstract