Paris saponin VII extracted from Trillium tschonoskii induces autophagy and apoptosis in NSCLC cells Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Shijing Qian, Shanshan Tong, Juan Wu, Lulu Tian, Zhan Qi, Beilei Chen, Deqiu Zhu, Yan Zhang Abstract Ethnopharmacological relevance Trillium tschonoskii Maxim, a perennial herb of the Trilliaceae, has been widely used to treat inflammation, hypertension and cancer. We investigated Paris saponin VII's (PS VII), isolated from Trillium tschonoskii Maxim, function in mediating autophagy and apoptosis in NSCLC cells. Materials and methods We treated various NSCLC cells with different concentrations of PS Ⅶ and then measure the cell apoptosis by using flow cytometry assays and western blot. Autophagy were investigated by using western blot, transmission electron microscopy and immunofluorescence analysis. We also use a xenograft model of nude mice to measure the effect of PS Ⅶ in vivo. Results Treatment with PS Ⅶ significantly inhibit NSCLC cell growth, especially for A549 (IC50 = 1.53 μM). Moreover, PS VII induces caspase-dependent apoptosis and autophagy through AMPK-ULK1 pathway. After blocking autophagy by 3-methyladenine (3-MA), PS VII induced cell death was significantly increased. In vivo, the co-treatment with PS VII and 3-MA dramatically inhibited A549 tumor growth in immune deficient mice and has similar inhibition rates as cisplatin group. Conclusion Our results suggest that a combination of PS VII and autophagy inhibitor may be a potential anticancer strategy in the NSCLC therapy. Graphical abstract

Natural products against acute respiratory infections: Strategies and lessons learned Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Julia Langeder, Ulrike Grienke, Ya Chen, Johannes Kirchmair, Michaela Schmidtke, Judith M. Rollinger Abstract Ethnopharmacological relevance A wide variety of traditional herbal remedies have been used throughout history for the treatment of symptoms related to acute respiratory infections (ARIs). Aim of the review The present work provides a timely overview of natural products affecting the most common pathogens involved in ARIs, in particular influenza viruses and rhinoviruses as well as bacteria involved in co-infections, their molecular targets, their role in drug discovery, and the current portfolio of available naturally derived anti-ARI drugs. Materials and methods Literature of the last ten years was evaluated for natural products active against influenza viruses and rhinoviruses. The collected bioactive agents were further investigated for reported activities against ARI-relevant bacteria, and analysed for the chemical space they cover in relation to currently known natural products and approved drugs. Results An overview of (i) natural compounds active in target-based and/or phenotypic assays relevant to ARIs, (ii) extracts, and (iii) in vivo data are provided, offering not only a starting point for further in-depth phytochemical and antimicrobial studies, but also revealing insights into the most relevant anti-ARI scaffolds and compound classes. Investigations of the chemical space of bioactive natural products based on principal component analysis show that many of these compounds are drug-like. However, some bioactive natural products are substantially larger and have more polar groups than most approved drugs. A workflow with various strategies for the discovery of novel antiviral agents is suggested, thereby evaluating the merit of in silico techniques, the use of complementary assays, and the relevance of ethnopharmacological knowledge on the exploration of the therapeutic potential of natural products. Conclusions The longstanding ethnopharmacological tradition of natural remedies against ARIs highlights their therapeutic impact and remains a highly valuable selection criterion for natural materials to be investigated in the search for novel anti-ARI acting concepts. We observe a tendency towards assaying for broad-spectrum antivirals and antibacterials mainly discovered in interdisciplinary academic settings, and ascertain a clear demand for more translational studies to strengthen efforts for the development of effective and safe therapeutic agents for patients suffering from ARIs. Graphical abstract

Paronychia argentea Lam. protects renal endothelial cells against oxidative injury Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): L. Arkoub-Hamitouche, V. González-del-Campo, M.E. López-Oliva, B. Fatiha, O.M. Palomino Abstract Ethnopharmacological relevance Paronychia argentea Lam. (Arabic tea), a species spontaneously growing in the Mediterranean area, has been used in folk medicine for renal diseases. Aim of the study To assess the antioxidant and protective potentials of different extracts from P. argentea in the renal endothelial NRK-52E cell line by several in vitro models, including a H2O2-induced oxidative stress model. Material and methods Aerial parts of P. argentea were collected in Algeria and ethanolic, chloroform and aqueous-chloroform extracts were obtained from dried plant. The antioxidant capacity was first evaluated by the Oxygen Radical Absorbance Capacity (ORAC) and the free radical scavenging activity (DPPH) methods. Cellular viability was assessed by MTT method assay after 24 h pretreatment with each extract concentration in order to measure protection from H2O2 in NRK-52E cells. Furthermore, the intracellular ROS formation (DCFH-DA method), was determined. Results P. argentea showed in vitro antioxidant activity as evidenced by the ORAC and DPPH assays. No cell toxicity was observed for concentrations ranging from 0.1 to 100 μg/mL of each extract. These extracts also exerted a protective effect on renal endothelial cells simultaneously treated with 1 mM H2O2. Chemical composition for the aqueous-chloroform extract was assessed by HPLC, as it showed the strongest antioxidant ability, revealing three quercetin derivatives as the main phenolic compounds. Conclusion P. argentea is endorsed with antioxidant activity and protects renal endothelial cells against oxidative damage which indicate this plant constitutes a potential treatment for renal diseases. Graphical abstract

Ethno-medicinal study of Artemisia ordosica Krasch. (traditional Chinese/Mongolian medicine) extracts for the treatment of allergic rhinitis and nasosinusitis Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Bin Xiao, Jin-Hua Wang, Cheng-Yan Zhou, Jun-Miao Chen, Na Zhang, Na Zhao, Xiao-Yan Han, Yi-Xuan Niu, Yu-Bao Feng, Guan-Hua Du Abstract Ethnopharmacological relevance Artemisia ordosica Krasch. (AOK) has been used for rheumatic arthritis, cold headache, sore throat, etc. in traditional Chinese/Mongolian medicine and is used for nasosinusitis by local Mongolian “barefoot” doctors. Up to now, their mechanisms are still unclear. Aim To evaluate the in vivo anti-inflammatory and allergic rhinitis (AR) alleviating effect as well as in vitro antimicrobial activities of AOK extracts to verify its ethno-medicinal claims. Materials and methods Crude extracts (methanol/95%-ethanol/ethyl acetate) of AOK root/stem/leaf and fractions (petroleum ether/ethyl acetate/n-butanol/aqueous) of AOK root extract were prepared. Xylene-induced ear swelling model in mouse and ovalbumin (OVA)-induced AR model in guinea pig were established. Ear swelling degrees of mice were measured. The numbers of rubbing movement and sneezes of guinea pigs were counted to evaluate the symptoms of AR. The serum levels of histamine, INF-γ, IL-2/4/10, and VCAM-1 were measured by ELISA assay. The histological changes of nasal mucosa were investigated by light microscope after H&E staining. Antimicrobial activities of AOK extracts were also tested. LC-MS/MS analysis was performed to characterize the constituents of active extract and molecular docking was conducted to predict the biological mechanism. Results In ear-swelling model, extract (100.00 mg/kg) from the ethyl acetate layer of 95% ethanol (100.00 mg/kg) showed better swelling inhibition in mice than positive control (dexamethasone, 191.91 mg/kg). In AR model, extract from the ethyl acetate layer of 95% ethanol significantly alleviated the AR symptoms in guinea pigs, decreased the serum levels of histamine, INF-γ, IL-2/4/10, and VCAM-1, and reduced the infiltration of eosinophil in nasal mucosa. For Staphylococcus aureus, the ethyl acetate extract of AOK stem showed the highest inhibition (MIC=1.25 mg/mL), for Escherichia coli, n-butanol layer of 95% ethanol extract of AOK root showed the highest inhibition (MIC=15.00 mg/mL), for Candida glabrata, 95%-ethyl acetate extract of AOK leaf showed the best inhibition (MIC=0.064 mg/mL), while ethyl acetate and n-butanol layers showed similar inhibition on MRSA (MIC=7.50 mg/mL). LC-MS/MS characterization showed that dicaffeoylquinic acids account for more than 30% of ethyl acetate layer of AOK extract. Dicaffeoylquinic acids bind with histamine-1 receptor with high affinities and interesting modes. Conclusions Extracts from AOK had interesting anti-inflammatory activity in mice, alleviating effect against OVA-induced AR in guinea pigs, and antimicrobial activities in vitro, which support the ethno-medicinal use of it. The main constituents in ethyl acetate layer of AOK root extract are dicaffeoylquinic acids and could bind with histamine-1 receptor well. These findings highlighted the importance of natural product chemistry study of AOK. Graphical abstract

Gastroprotective effects of Erythrina speciosa (Fabaceae) leaves cultivated in Egypt against ethanol-induced gastric ulcer in rats Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Nouran M. Fahmy, Eman Al-Sayed, Haidy E. Michel, Mohamed El-Shazly, Abdel Nasser B. Singab Abstract Ethnopharmacological relevance Members of the genus Erythrina have been traditionally used in the treatment of various ailments such as inflammation and gastrointestinal disorders. Erythrina speciosa (Fabaceae) is a spiny, deciduous shrub or small tree native to Southern America in Brazil. It is cultivated in Africa and Asia. The traditional usage of E. speciosa indicated its antibacterial, analgesic, and anti-inflammatory activities. Aim of the study Evaluation of the phytochemical constituents, gastroprotective effects and possible mechanism of action of the ethyl acetate fraction obtained from the methanol extract of E. speciosa leaves (ESLE). Materials and methods Chemical characterization of ESLE was done using high performance liquid chromatography coupled to mass spectrometry (HPLC-MS). The gastroprotective activity of ESLE was evaluated using ethanol-induced gastric-ulcer model in rats. Rats were pre-treated with ESLE 25, 50 and 100 mg/kg 1 h before the administration of absolute ethanol. Histological analysis, mucin content, and total acidity were evaluated. The possible mechanism of action of ESLE was studied through the examination of oxidative stress and inflammatory markers, PGE2, and NF-κB, iNOS, COX-2, and HSP-70 immunoexpression. In vitro, anti-Helicobacter pylori activity of ESLE was also studied using micro-well dilution method. Results Fourteen compounds were tentatively identified including alkaloids, flavonoids, and saponins. ESLE exerted a powerful gastroprotective effect. The pre-treatment with ESLE at different doses resulted in a significant reduction in gastric lesions and significant elevation in the mucin production. These effects could be partially mediated by the potent anti-inflammatory activity of ESLE as evidenced by the significant reduction in the immunoexpression of NF-κB, COX-2, iNOS and the reduction in the pro-inflammatory marker, TNF-α. ESLE counteracted the ethanol-induced oxidative stress by increasing the levels of depleted GSH and catalase as well as significantly attenuating the ethanol-induced lipid peroxidation tissue levels. In addition, ESLE exhibited in vitro antibacterial activity against H. pylori. Conclusions The chemical constituents of ESLE strongly support its potent gastroprotective effect suggesting its future potential application in the management of gastric ulcer by eliminating its symptoms and causes including H. pylori. Graphical abstract

Traditional Medicine (Mahuang-Tang) Improves Ovarian Dysfunction and the Regulation of Steroidogenic Genes in Letrozole-Induced PCOS Rats Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Hyun Yang, Young Ho Lee, Sang R. Lee, Pelin Kaya, Eui-Ju Hong, Hye Won Lee Abstract Ethnopharmacological relevance Mahuang-Tang (MHT) has traditionally been used in Asia to treat a variety of diseases, such as fever without sweating, joint pain, lower back pain, asthma, and gynecological conditions. Polycystic ovary syndrome (PCOS) is a kind of gynecological disease that causes amenorrhea, infertility, and menopausal and urogenital disorders that could benefit from MHT treatment. Aim of the study In this study, we examined the effects of MHT on ovarian hormones and steroidogenic enzymes in female PCOS rats. Methods and results The PCOS rat model was induced by Letrozole, and an in vivo evaluation of whether the dietary consumption of MHT improved the PCOS-like symptoms was conducted. The luteinizing hormone (LH) level and luteinizing hormone/follicular-stimulating hormone (LH/FSH) ratio increased in PCOS rats but decreased following MHT treatment. In the PCOS rats, the reduced estrogen level was restored to that of normal controls with MHT treatment in serum. The transcription level(s) of gonadotropin receptors (Fshr and Lhr), steroid receptors (Pgr, and Esr1) and steroidogenic enzymes (Cyp19a1, Hsd3b1, Hsd17a1, and Cyp11a1) changed under the PCOS condition, and were regulated by MHT treatment in the ovaries of PCOS rats. The reproductive tissues of Letrozole-induced PCOS rats were restored into estrogenic condition from androgen environments. Conclusion These results suggest that MHT ameliorates the symptoms of PCOS by improving the dysregulation of ovarian steroids and steroidogenic enzymes in PCOS rats. Graphical abstract

Dendrobium officinale polysaccharide ameliorates diabetic hepatic glucose metabolism via glucagon-mediated signaling pathways and modifying liver-glycogen structure Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Yage Liu, Linlin Yang, Yu Zhang, Xiaocui Liu, Zhijing Wu, Robert G. Gilbert, Bin Deng, Kaiping Wang Abstract Ethnopharmacological relevance Dendrobium officinale polysaccharide (DOP) is the main active ingredient of Dendrobium officinale Kimura & Migo, which is a precious traditional Chinese medicine and often used in treatment of hepatitis, diabetes, obesity and rheumatoid arthritis. Aim of the study: DOP exhibits significant hypoglycemic activity, while its mechanism remains unclear. The present study aims to investigate the hypoglycemic mechanisms of DOP based on the glucagon-mediated signaling pathways and the liver glycogen structure, which catalyze hepatic glucose metabolism, and provide new knowledge about the antidiabetic mechanism of DOP and further evidence for its clinical use for diabetes. Materials and methods DOP were obtained from the dry stems of Dendrobium officinale by water extraction and alcohol precipitation method. T2DM mice model was established by high-fat diet combined with streptozotocin. Liver histopathological changes were observed by H&E and PAS straining. Pancreatic histology was studied by H&E staining and immunofluorescence analysis. The levels of glucagon and insulin were detected by Elisa Kit and the hepatic glycogen content was detected by GOPOD. The expressions of the hepatic glycogen-related metabolism enzymes, hepatic gluconeogenesis enzymes, and the related protein in cAMP-PKA and Akt/FoxO1 signaling pathways were detected by western blots. Liver glycogen was extracted from the liver tissues by sucrose density gradient centrifugation, and size exclusion chromatography (SEC) was used to analyze the structure of liver glycogen. Results DOP could significantly affect the glucagon-mediated signaling pathways, cAMP-PKA and Akt/FoxO1, to further promote hepatic glycogen synthesis, inhibit hepatic glycogen degradation and hepatic gluconeogenesis. Moreover, DOP could reverse the instability of the liver glycogen structure and thus probably suppressed glycogen degradation. Thus, DOP finally would ameliorate hepatic glucose metabolism via glucagon-mediated signaling pathways and modifying liver-glycogen structure in diabetic mice. Conclusions The hypoglycemic mechanism of DOP might be associated with the regulation of glucagon-mediated hepatic glycogen metabolism and gluconeogenesis, and of liver glycogen structure, contributing to improved hepatic glucose metabolism in diabetic mice. Graphical abstract DOP could regulate glucagon-mediated signaling pathways and reverse the instability of liver glycogen structure, contributing to improved hepatic glucose metabolism.

Toxicological safety evaluation in acute and 28-day studies of aqueous extract from Bei-Qi-Wu-Jia formula Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Liutao Zhao, Pan Li, Hongde Xu, Bingqian Han, Jingjing Chen, Ziqing Gao, Jianglong Li, Xianbin Li, Chunli Wu Abstract Ethnopharmacological relevance Bei Qi Wu Jia (BQWJ), a modern preparation of a traditional Chinese medicinal formula, is a combination of Radix Astragali and Acanthopanacis Senticosi. Although BQWJ has been used to treat insomnia, fatigue, and loss of appetite, toxicological safety studies are rare in the literature. Aim of the study: To evaluate the acute and subacute toxicity of BQWJ extract after oral administration in mice and rats, respectively. Materials and methods In the acute toxicity study, mice underwent oral administration of 67.5 g extract/kg/day. In the subacute toxicity study, rats underwent a single oral administration of 1.25, 2.5, 5.0, or 10.0 g/kg/day of BQWJ extract for 28 days. The animals’ general behavior, body weight, food intake, biochemical and hematologic parameters, organ coefficients, and pathological morphology were analyzed. Results No evidence of toxicity was observed in the mice after acute exposure to BQWJ extract. The subacute results included no deaths and no changes in general behavior. Although BQWJ extract resulted in some significant changes in other parameters, these alterations cannot be considered treatment-related because they remained within normal ranges throughout the 28 days. Conclusions In conclusion, the oral administration of BQWJ extract at doses of less than 67.5 g/kg/day for 1 day or 10.0 g/kg/day for 28 consecutive days can be considered safe and showed no distinct toxicity or side effects in this study. Graphical abstract The mixture of Radix Astragali and Acanthopanacis Senticosi (ratio 1:1.12) was powdered and extracted in boiling water two times under reflux condition for 3 h each time. The two decoctions were mixed and filtered and then concentrated under reduced pressure, yielding Bei Qi Wu Jia extract, and stored at room temperature. Strict quality control was established by high-performance liquid chromatography (HPLC) analysis using two methods.In the acute toxicity study, mice underwent oral administration of 67.5 g/kg/day of Bei Qi Wu Jia extract. In the subacute toxicity study, rats underwent a single oral administration of 1.25, 2.5, 5.0, or 10.0 g/kg/day of Bei Qi Wu Jia extract for 28 days. The animals’ general behavior, body weight, food intake, biochemical and hematologic parameters, organ coefficients, and pathological morphology were analyzed.

Polyalthia longifolia leaves methanolic extract targets entry and budding of viruses-an in vitro experimental study against paramyxoviruses Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Prashant Yadav, Soumen Choudhury, Sanjay Barua, Nitin Khandelwal, Naveen Kumar, Amit Shukla, Satish K. Garg Abstract Ethnopharmacological relevance Synthetic antiviral drugs have several limitations including high cost. Thus research on antiviral property of medicinal plants is continuously gaining importance. Polyalthia longifolia possesses several medicinal properties and has been used in traditional ayurvedic medicine for treatment of dermatological ailments as kushta, visarpa/herpes virus infection and also to treat pyrexia of unknown origin as mentioned in Visarpa Chikitsa. Aim of the study Keeping in view the cytotoxic, anti-cancer activity and antiviral efficacy of Polyalthia longifolia against herpes, present study was undertaken to evaluate the in vitro antiviral activity of methanolic extract of Polyalthia longifolia leaves, if any, and to unravel the possible target(s)/mechanism of action. Material and methods Antiviral activity of Polyalthia longifolia methanolic extract was studied using Vero cell lines against paramyxoviruses, namely-peste des petits ruminants virus (PPRV) and Newcastle disease virus (NDV). Cytotoxicity of the test extract was evaluated employing MTT assay. Virucidal activity, and viral-attachment, virus entry and release assays were determined in Vero cells using standard experimental protocols. The viral RNA in the virus-infected cells was quantified by qRT-PCR. Results At non-cytotoxic concentration, methanolic extract of Polyalthia longifolia leaves was found to inhibit the replication of PPRV and NDV at viral entry and budding level, whereas other steps of viral life cycle such as attachment and RNA synthesis remained unaffected. Conclusions Polyalthia longifolia leaves extract possesses promising antiviral activity against paramyxoviruses and acts by inhibiting the entry and budding of viruses; and this plant extract evidently possesses excellent and promising potential for development of effective herbal antiviral drug. Graphical abstract

Antinociceptive effects of Salvia divinorum and bioactive salvinorins in experimental pain models in mice Publication date: 10 February 2020 Source: Journal of Ethnopharmacology, Volume 248 Author(s): Lorenzo Leonel Tlacomulco-Flores, Myrna Déciga-Campos, María Eva González-Trujano, Azucena Ibeth Carballo-Villalobos, Francisco Pellicer Abstract Ethnopharmacological relevance Salvia divinorum Epling & Játiva is a Mexican plant used not only in rituals but also in traditional medicine for pain relief. One of the most known bioactive compounds is salvinorin A, which acts centrally in kappa-type opioid receptors. Aim of the study Despite its traditional use as a medicinal plant, there is not enough scientific investigation to reinforce its potential as analgesic. In this study, Salvia divinorum antinociceptive activity was evaluated in experimental models of nociceptive pain; the writhing test and formalin-induced licking behavior in mice. Material and methods Different Salvia divinorum extracts were prepared by maceration at room temperature in increased polarity (hexane, ethyl acetate and methanol). The ethyl acetate extract (EAEx) was chosen in order to be fractioned and to obtain a mixture of salvinorins. The antinociceptive effect of EAEx (3, 10, 30, and 100 mg/kg, i.p.) was compared with that of tramadol (a partial opioid agonist analgesic drug, 30 mg/kg, i.p.) and the mixture of salvinorins (30 mg/kg, i.p.). In addition, a participation of opioids (naloxone, NX 1 and/or 3 mg/kg, i.p.) and serotonin 5-HT1A receptors (WAY100635, 0.32 mg/kg, i.p.) was investigated as possible inhibitory neurotransmission involved. Results As a result, the EAEx produced significant and dose-dependent antinociceptive effect concerning salvinorins constituents. This effect was blocked in the presence of NX and WAY100635 in the abdominal test, but only by NX in the formalin-induced licking behavior. Whereas, the effect of salvinorins mixture involved opioids and serotonin 5-HT1A receptors. Conclusion Data provide evidence of the potential of this species, where salvinorin A is in part responsible bioactive constituent involving participation of the opioids and/or 5-HT1A serotonin receptors depending on the kind of pain model explored. Graphical abstract