Exploring the key anti-fatigue components of American ginseng based on metabolomics and zebrafish models / 药学学报

The aim of this paper is to explore the key anti-fatigue active components in the saponin-like composition of American ginseng. The anti-fatigue activity of western ginseng samples was evaluated using a zebrafish model; metabolomics techniques were used to identify the main saponins in western ginseng from different origins; the active substances and relevant targets of the anti-fatigue effect of western ginseng were initially screened by constructing a PPI protein interaction network between western ginseng saponins and disease targets, and the key active ingredients were screened using a molecular docking method; finally, the anti-fatigue activity of the key active ingredients was evaluated using a zebrafish, animal experiment was approved by the Ethics Committee of Shandong Academy of Medical Sciences (SYXK20220005). The anti-fatigue activity of the key active ingredients was evaluated using a zebrafish model. The results of the zebrafish activity evaluation showed that there were significant differences in the activities of the western ginseng samples from the two origins, and a total of 10 different saponins were identified as possibly related to the anti-fatigue activity after further metabolomic testing and pattern discrimination. The core anti-fatigue targets were screened with the help of component-disease target PPI, combined with pharmacophore-like parameters and molecular docking techniques, and pseudoginsenoside F11 was found to have good binding activity to five of the targets. Finally, the zebrafish model revealed that pseudoginsenoside F11 exhibited significant anti-fatigue activity. This study used metabolomics and zebrafish model to screen the key active substances of pseudoginsenoside F11 for its anti-fatigue activity, which will provide a reference for further research on the anti-fatigue of pseudoginsenosides.

Application of UPLC-QTOF-MS non-targeted metabonomics in mechanism study of property differences of ginseng and American ginseng / 中国中药杂志

This study adopted ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF-MS)-based untargeted metabolomic approaches for exploring the changes in endogenous metabolites of rat serum related to property differences between ginseng and American ginseng. Then the action mechanisms of them with warm and cool properties and the effects of processing on their property changes were investigated. Based on principal component analysis(PCA), the differences in metabolite profiles between ginseng, red ginseng, American ginseng, and red American ginseng were compared. After that, 16 potential differential endogenous biomarkers were identified by orthogonal partial least squares discriminant analysis(OPLS-DA) and online database searching. And the related metabolic pathways were systematically analyzed. By comparing content variations of these 16 potential differential endogenous biomarkers, we have found that 10 potential differential biomarkers were responsible for the warm property of ginseng and red ginseng, and 9 were related to the cool property of American ginseng and red American ginseng. As demonstrated by in-depth analysis of related metabolic pathways of differential biomarkers, ginseng and American ginseng mainly played a role in regulating the energy metabolism of amino acid, glycolysis, and fatty acids, during which they exhibited differences in property. The comparison of content variations of these differential endogenous between groups revealed that the energy metabolism of red ginseng group was stronger than that of ginseng group, consistent with the traditional processing theory that the warming and tonifying effects of ginseng could be enhanced after processing. The property of red American ginseng was similar to that of American ginseng, both cool in property, but American ginseng was cooler than red American ginseng. It can be seen that non-targeted metabolomic approaches can be utilized to study mechanisms underlying property differences of Chinese medicines and the effects of processing on their property changes.

Dendrobium officinale Kimura et Migo and American ginseng mixture: A Chinese herbal formulation for gut microbiota modulation / 中国天然药物

Dendrobium officinale Kimura et Migo (D. officinale) is a famous traditional Chinese medicine (TCM). A mixture of D. officinale and American ginseng has been shown to enhance cell-mediated immunity, humoral immunity, and monocyte/macrophage functions in mice. Here, the effects of a D. officinale and American ginseng mixture on the structure of gut microbial community in dogs were examined using high-throughput 16S rRNA gene amplicon sequencing. The data revealed that while the mixture did not change the diversity of gut microbial community significantly, differences among individuals were significantly reduced. Furthermore, the mixture-responsive operational taxonomic units (OTUs) exhibited a phase-dependent expression pattern. Fifty-five OTUs were found to exhibit a mixture-induced expression pattern, among which one third were short-chain fatty acid (SCFA)-producing genera and the others were probiotic genera included Lactobacillus spp., Sutterella, Alistipes, Anaerovorax, Bilophila, Coprococcus, Gordonibacter, Oscillibacter, among others. By contrast, 36% of the OTUs exhibiting a mixture-repressed expression pattern were disease-associated microorganisms, and six genera, namely Actinomyces, Escherichia/Shigella, Fusobacterium, Slackia, Streptococcus and Solobacterium, were associated with cancer. In addition, five genera were closely associated with diabetes, namely Collinsella, Rothia, Howardella, Slackia and Intestinibacter. Our results indicate that this D. officinale and American ginseng mixture may be used as a prebiotic agent to enhance SCFA-producing genera and prevent gut dysbiosis.

Effect of American ginseng saponins on enhancing immunity based on zebrafish model organisms / 中草药

Objective: To study the immune enhancing effect of saponins from American ginseng and its mechanism. Methods: The drug dosage of American ginseng saponin extract and the optimal modeling dose of rapamycin were determined by tolerance test and immunodeficiency model establishment experiments. Zebra fish were divided into control group (0.5% dimethyl sulfoxide), model group (4 μg/mL rapamycin), and three origins American ginseng treatment groups with different concentrations (Shandong, Northeast, Canada, 4 μg/mL rapamycin +10, 25 and 50 μg/mL American ginseng saponin extract) 48 h after fertilization (48 hpf). After a certain period of incubation, the number of neutrophils in the tail of zebrafish, macrophage phagocytosis, and interferon gamma (IFN-γ) content in the body were used as indexes, and the growth rate of neutrophils was calculated to investigate the immune-enhancing effect of American ginseng saponins. Results: When the dosage of American ginseng saponins was higher than 50 μg/mL, the mortality of Zbra-fish increased with the increase of the concentration and the time of administration; Compared with control group, in model group, the number of neutrophils in the tail of zebrafish was decreased significantly (P < 0.01), and the content of IFN-γ in the body was decreased significantly (P < 0.01); Compared with model group, in 10 μg/mL and 25 μg/mL American ginseng treatment group, the number of neutrophils in the tail of zebrafish was increased significantly (P < 0.01), the number of macrophages engulfing ink particles was increased significantly (P < 0.01), the content of IFN-γ in vivo was increased significantly (P < 0.01), and the growth rate of neutrophils was between 19.73% and 96.49%. Conclusion: The American ginseng saponins have a better effect on enhancing immunity.

Prediction of the globally ecological suitability of Panax quinquefolius by the geographic information system for global medicinal plants (GMPGIS) / 中国天然药物

American ginseng (Panax quinquefolius L.) is a well-known Asian traditional herbal medicine with a large market demand. The plant is native to eastern North America, and its main producing areas worldwide are decreasing due to continuous cropping obstacles and environmental changes. Therefore, the identification of maximum similarities of new ecological distribution of P. quinquefolius, and prediction of its response to climate change in the future are necessary for plant introduction and cultivation. In this study, the areas with potential ecological suitability for P. quinquefolius were predicted using the geographic information system for global medicinal plants (GMPGIS) based on 476 occurrence points and 19 bioclimatic variables. The results indicate that the new ecologically suitable areas for P. quinquefolius are East Asia and the mid-eastern Europe, which are mainly distributed in China, Russia, Japan, Ukraine, Belarus, North Korean, South Korea, andRomania. Under global climate change scenarios, the suitable planting areas for P. quinquefolius would be increased by 9.16%-30.97%, and expandingnorth and west over the current ecologically suitable areas by 2070. The potential increased areas that are ecologically suitable include northern Canada, Eastern Europe, and the Lesser Khingan Mountains of China, and reduced regions are mainly in central China, the southern U.S., and southern Europe. Jackknife tests indicate that the precipitation of the warmest quarter was the important climatic factor controlling the distribution of P. quinquefolius. Our findings can be used as auseful guide for P. quinquefolius introduction and cultivation in ecologically suitable areas.

Extraction Technology Optimization of the Total Polysaccharides and Saponins from American Ginseng / 中国药师

Objective: To optimize the extraction method for American ginseng. Methods: Based on the contents of the total poly-saccharides and saponins in American ginseng, the ethanol concentration, extraction duration, solid-liquid ratio and extraction time were selected to determine the factors and levels by single-factor test. And then 3 factors further considered were the concentration of ethanol, solid-liquid ratio and extraction time. Results: The optimum extraction process was adding 12-fold amount of 40% ethanol, and extracting for 3 times. Conclusion: The optimized extraction process is simple, quick and stable, and can be used to extract the total polysaccharide and saponins from American gingeng.

Evaluation and prediction of ecological suitability of medicinal plant American ginseng (panax quinquefolius) / 中草药·英文版

Objective: American ginseng is a medicinal plant with large market demands, however, its producing areas are shrinking because of the continuous cropping obstacles in China. Therefore, it is urgent to establish a suitable model to determine the new producing areas. Here we evaluated and predicted the suitable areas of American ginseng using the maximum entropy model (MaxEnt). Methods: Based on the 37 environmental variables over thirty years from 1970 to 2000 and 226 global distribution points of American ginseng, MaxEnt was used to determine the global ecological suitable areas for American ginseng. The Receiver Operating Curve (ROC) was used to evaluate the model prediction accuracy. Meanwhile, an innovative ecological variable, the precipitation–temperature ratio, was established to indicate the climate characteristic in the American ginseng suitable areas based on the monthly precipitation and temperature. Results: The potential ecological suitable areas of American ginseng were primarily in Appalachian Mountain in America and Changbai Mountain in China, about in the range of 35°N–50°N, 60°W–120°W and 35°N–50°N, 110°E–145°E, respectively, including the United States, Canada, China, North Korea, South Korea, Russia and Japan. South Korea and Japan were the potential producing regions. The precipitation–temperature ratios were stable at (0.22, 0.56) of the vigorous growth period (April–October) in the best suitable areas of American ginseng, serving as characteristic parameters to optimize the prediction model. The model showed that the common soil parameters were pH 4.5–7.2, Base Saturation (BS) above 80%, Cation Exchange Capacity (CEC) 10–20 cmol/kg, organic carbon (OC) < 1.4%, and the soil types were sandy loam or loam. Conclusion: An optimized MaxEnt model was established to predict the producing area for American ginseng that needed to be validated by a field test.

Effect of Ginseng on Blood Pressure: A Systematic Review and Meta-Analysis

OBJECTIVE: A meta-analysis was performed to determine effect of ginseng on blood pressure. METHODS: The databases of PubMed, Embase, Cochrane Library, RISS, DBpia, KISS, and Koreamed were searched for all published studies from inception to January 2016. The following terms were used: "ginseng", "hypertension", and "blood pressure". Using the Review Manager 5, mean differences (MDs) were pooled to measure the effect of ginseng on blood pressure compared to that of placebo. RESULTS: Eleven randomized controlled trials were included. In this meta-analysis, ginseng treatment significantly lowered systolic blood pressure (SBP) in a dose-independent way (MD: -1.99, p = 0.04). In subgroup analysis, 8-12 week consumption of ginseng achieved significantly greater reduction in SBP (MD: -3.14, p = 0.03), while single administration of ginseng failed to show BP-lowering effect. When ingested over 8-12 weeks, ginseng significantly lowered diastolic blood pressure (DBP) (MD: -1.96, p = 0.03). No significant association was found between ginseng dose and the magnitude of BP-lowering effect. However, a significant positive relationship was observed between baseline SBP level and the magnitude of SBP reduction (r = 0.848, p = 0.033). Such a relationship was not seen in DBP. CONCLUSION: Consumption of ginseng for 8-12 weeks achieved significant reductions in SBP and DBP in a dose-independent way. There was a significant positive relationship between baseline SBP level and the magnitude of SBP reduction.

Quality evaluation of American ginseng using UPLC coupled with multivariate analysis / 中国中药杂志

An ultra performance liquid chromatography (UPLC)method combined with multivariate data analysis was developed to evaluate the quality of American ginseng by simultaneously determining the concentrations of six ginsenosides (Rg₁, Re, Rb₁, Rc, Ro and Rd)in the samples. For UPLC, acetonitrile with 0.01% formic acid and water with 0.01% formic acid were used as the mobile phase with gradient elution. Under the established chromatographic conditions, the six ginsenosides could be well separated and the results of linearity, stability, precision, repeatability, and recovery rate all reached the requirement of quantification analysis, respectively. The total contents of Rg₁, Re, and Rb₁ in 57 samples all reached the requirement of the 2015 edition of Chinese Pharmacopoeia. At the same time, the experimental data were analyzed by principle component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). The crude drugs and the decoction pieces can be discriminated by a PCA method and the samples with different age can be distinguished by a PLS-DA method.

Advances in Transcriptomic Studies and Ginsenoside Biosynthesis of American Ginseng / 中草药·英文版

American ginseng (. Panax quinquefolius L.), belonging to the Araliaceae family, is one of the most widely used traditional herbs in the world. Its major bioactive constituents are triterpene saponins known as ginsenosides. Up to date, it is still a big challenge to sequence and assemble the large and repeat-enriched genome of tetraploid American ginseng, using whole genome shotgun (WGS) sequencing strategy. The lack of American ginseng genome information has significantly impeded its genetic and functional genomic studies. With the development of next-generation sequencing (NGS) technologies, sequencing and analysis of transcriptomes have become powerful tools for the discovery of novel genes and elucidation of specific biosynthetic pathways of secondary metabolites. Here we summarized the recent advances in the transcriptomic studies of American ginseng, including high-throughput transcriptome sequencing, assembly, and functional gene annotation and classification. Based on the results of transcriptomic data mining and co-expression analyses, many candidate genes possibly involved in the biosynthetic pathway of ginsenosides have been found, thereby providing an unparalleled opportunity to fully understand the mechanism of ginsenoside biosynthesis and its regulations in American ginseng. Advances in transcriptomic studies will contribute to the molecular breeding and planting management of American ginseng and to the development of novel ginsenoside-type drugs.

RAPD and ISSR analyses of genetic diversity of American ginseng germplasm from different habitats in China / 中草药

Objective: To analyze the genetic diversity of American ginseng (Panax quinquefolium) produced in 10 regions of China by usting RAPD and ISSR. Methods: Genomic DNA was extracted by CTAB and the following two kinds of ginseng were used as controls which are ginseng (P. ginseng) produced in China and American ginseng produced in Canada. Thirteen RAPD primers and 12 ISSR primers were selected to perform PCR amplification. According to the band number, NTSYS-pc2.10e software was applied to the cluster analysis using UPGMA. Results: Thirteen RAPD primers had amplified 97 clear bands, 81 polymorphic bands, and the percentage of polymorphism was 85.51%; Twelve ISSR primers had amplified 99 clear bands, 64 polymorphic bands, and the percentage of polymorphism was 64.65%; Through cluster analysis, the samples were clustered into four categories by RAPD and RAPD + ISSR and two categories by ISSR. Conclusion: RAPD and ISSR markers are used to construct a dendrogram of samples, which is slightly different in classification, but the overall trend is consistent. Ginseng and American ginseng are clear distinction. On the ISSR, American ginseng from Xingshen Town and Beigang Town in Jilin province are gathered for a major categories with ginseng. In the growing environment and planting conditions, the genetic diversity of American ginseng has been changed in the part of northeast China compared with Canada.

Effects of Triterpenoid Glycosides from Fresh Ginseng Berry on SW480 Human Colorectal Cancer Cell Line / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The pharmacological activities, notably the anticancer properties, of bioactive constituents fromfresh American ginseng berry have not yet been well studied. In this study, we investigated the antiproliferative effects of fresh American ginseng berry extract (AGBE) and its representative triterpenoid glycosides using the human colorectal cancer cell line SW480. MATERIALS AND METHODS: Using high performance liquid chromatography (HPLC), the contents of 8 ginsenosides in AGBE were determined. The cell growth inhibitory effects of AGBE and three triterpenoid glycosides (ginsenosides Rb3, Re, and Rg3) were evaluated by proliferation assay and 3H-thymidine incorporation assay. Cell cycle and apoptotic effects were analyzed by using flow cytometry after staining with propidium iodide and annexin V. RESULTS: HPLC analysis data showed that AGBE has a distinct ginsenoside profile. AGBE inhibited SW480 cell growth significantly in a time-dependent (24-96 hours) and concentration-dependent (0.1-1.0 mg/mL) manner. Ginsenosides Rb3, Re, and Rg3 also possess significant antiproliferative activities on SW480 cells. 3H-thymidine incorporation assay indicated that AGBE and ginsenosides Rb3, Re, and Rg3 might inhibit the transferring and duplication of DNA in SW480 cells. Flow cytometric assay data suggested that AGBE arrested SW480 cells in S and G2/M phases, and significantly induced cell apoptosis. CONCLUSION: AGBE and ginsenosides Rb3, Re, and Rg3 possessed significant antiproliferative effects and induced changes of morphological appearance on SW480 cells. The mechanisms of the antiproliferation of AGBE and tested ginsenosides involved could be cell cycle arrest and induction of apoptosis.

American Ginseng Transcriptionally Activates p21 mRNA in Breast Cancer Cell Lines

American ginseng (AG) has been demonstrated to inhibit breast cancer cell growth in vitro. p21 protein, a universal cell cycle inhibitor, binds cyclin-CDK complexes, an important mechanism in cell cycle regulation. The purpose of this investigation was to determine if AG induces p21 gene expression in hormone sensitive (MCF-7) and insensitive (MDA-MB-231) breast cancer cell lines. Cells grown in steroid stripped medium (SSM) were treated with AG, 17-beta-estradiol (E2), genistein or cycloheximide (CHX). Northern blot analyses were performed using human p21Cip1 and 36B4 cDNA probes. Cell lines were transiently transfected with select mouse p21 CAT reporter constructs, including those lacking a p53 binding site. Cell cycle analyses was performed by FACScan. The results revealed that AG induced p21 mRNA expression in MCF-7 and MDA-MB-231 cells (p=0.0004; p< or =0.0001, respectively). Neither E2 nor genistein alter p21 mRNA expression. CHX, a protein synthesis inhibitor, did not block p21 mRNA expression induced by AG, indicating that p21 is induced as an immediate early gene. AG activated p21 reporter constructs in transfected cells, independent of p53 binding sites. The cell cycle proliferative phase was significantly decreased by AG and increased by E2 (p< or =0.0001). AG may inhibit breast cancer cell growth by transcriptional activation of the p21 gene, independent of p53.

Prospect of studies on four natures of ginseng,red ginseng,and American ginseng based on expression of apoptosis / 中草药

To study on the four natures (cold, hot, warm, and cool) of Chinese materia medica (CMM) is a central and knotty issue in the field of traditional Chinese medicine (TCM) and no substantial progress has been made so far, which is an obstacle in the course for the modernization of TCM. By application of apoptosis for researching the four natures of CMM, the relationship between the biological efficiency and apoptosis has been explained in a qualitative and quantitative way. A pattern recognition system for the four natures of CMM has been set up in a molecular level. That is a breakthrough to measure the four natures of CMM. Based on the abundant literatures at home and abroad, and the study done as well, the original thinking and fresh viewpoint on the orientation and development for the four natures of CMM have been put forward and then the explanation and prospective analysis on them have been carried out.