Gentianella acuta-derived Gen-miR-1 suppresses myocardial fibrosis by targeting HAX1/HMG20A/Smads axis to attenuate inflammation in cardiac fibroblasts.

BACKGROUND: Continuous activation and inflammation of cardiac fibroblasts (CFs) are essential for myocardial fibrosis. Gentianella acuta (Michx.) Hiitonen (G. acuta), that contains xanthones with cardioprotective properties, a typical healthful herb extensively used to treat cardiovascular diseases in Inner Mongolia region of China. However, it remains unknown whether or not G. acuta-derived miRNAs can shield CFs from activation by inflammatory stimulation. Therefore, we tend to investigated the role and core mechanism of G. acuta-derived Gen-miR-1 in regulating fibrosis and inflammation induced by TGF-ß1. METHODS: An animal model for myocardial infarction was built by subcutaneous injections of ISO and treated with Gen-miR-1 using intragastric administration. The protective effect of Gen-miR-1 on the heart was assessed by pathomorphological analysis of myocardial fibrosis. Using loss- and gain-of-function approaches, Gen-miR-1 regulation of HAX1/HMG20A/Smads axis was investigated by utilizing luciferase assay, Western blot, co-immunoprecipitation, etc. RESULTS: Screened and identified Gen-miR-1 from G. acuta. Gen-miR-1 can enter the mouse body, and markedly inhibit myocardial infarction induced by ISO in mice, as well as suppresses fibrosis in CFs and attenuates the inflammatory response elicited by TGF-ß1 in vitro. Gen-miR-1 downregulates HCLS1-related Protein X-1 (HAX1) expression through direct binding to the 3' UTR of HAX1, which in turn relieves HAX1 from promoting the expression of high-mobility group protein 20A (HMG20A), whereas HMG20A downregulation restrains the activation of TGF-ß1/Smads signaling pathways, subsequently resulting in a decrease of fibrosis and in facilitating CFs anti-inflammatory effects induced by Gen-miR-1 in the context of CFs activation induced by TGF-ß1. CONCLUSIONS: Our results first uncovered unique bioactive components in G. acuta and elucidated the molecular mechanism by which G. acuta-derived Gen-miR-1 suppress inflammation and myocardial fibrosis. These findings expand our understanding of G. acuta's therapeutic properties and bioactive constituents. Gen-miR-1-regulated HAX1/HMG20A/Smads axis will be one potential therapeutic target for cardiac remodeling.

Xanthones from Gentianella acuta (Michx.) Hulten Ameliorate Colorectal Carcinoma via the PI3K/Akt/mTOR Signaling Pathway.

Colorectal carcinoma (CRC) is a kind of malignant tumor closely related to ulcerative colitis. Xanthone derivatives are one of the most promising therapeutic drugs which have been used in phase I/II clinical trials for cancer therapy. Our previous study indicated that the aerial parts of Gentianella acuta Michx. Hulten (GA) was rich in xanthones and showed a good therapeutic effect on ulcerative colitis in mice, suggesting that GA xanthones might have some therapeutic or ameliorative effects on CRC. However, no relevant study has been reported. This study aims to find the effective substances of GA inhibiting CRC and clarify their mechanism. Solvent extraction, column chromatographic separation, and LC-MS analysis were used to characterize the 70% EtOH extract of GA and track xanthones abundant fraction XF. MTT assay was carried out to clarify the activity of GA fractions; the result showed XF to be the main active fraction. LC-MS analysis was executed to characterize XF, 38 xanthones were identified. Network pharmacology prediction, in vitro activity screening, and molecular docking assay were combined to predict the potential mechanism; the PI3K/Akt/mTOR signaling pathway was found to be most important. Western blot assay on the main active xanthones 1,3,5-trihydroxyxanthone (16), 1,3,5,8-tetrahydroxyxanthone (17), 1,5,8-trihydroxy-3-methoxyxanthone (18), and 1,7-dihydroxy-3,8-dimethoxyxanthone (19) was used to verify the above prediction; these xanthones were found to inhibit the PI3K/Akt/mTOR signaling pathway, and 17 played a significant role among them through Western blot assay using PI3K/AKT/mTOR agonist IGF-1. In conclusion, this study demonstrated that GA xanthones were effective compounds of GA inhibiting CRC by regulating PI3K/Akt/mTOR signaling pathway transduction, at least. Importantly, 1,3,5,8-tetrahydroxyxanthone (17), the most abundant active xanthone in GA, might be a candidate drug for CRC.

Gentianella acuta improves TAC-induced cardiac remodelling by regulating the Notch and PI3K/Akt/FOXO1/3 pathways.

Cardiac remodelling mainly manifests as excessive myocardial hypertrophy and fibrosis, which are associated with heart failure. Gentianella acuta (G. acuta) is reportedly effective in cardiac protection; however, the mechanism by which it protects against cardiac remodelling is not fully understood. Here, we discuss the effects and mechanisms of G. acuta in transverse aortic constriction (TAC)-induced cardiac remodelling in rats. Cardiac function was analysed using echocardiography and electrocardiography. Haematoxylin and eosin, Masson's trichrome, and wheat germ agglutinin staining were used to observe pathophysiological changes. Additionally, real-time quantitative reverse transcription polymerase chain reaction and western blotting were used to measure protein levels and mRNA levels of genes related to myocardial hypertrophy and fibrosis. Immunofluorescence double staining was used to investigate the co-expression of endothelial and interstitial markers. Western blotting was used to estimate the expression and phosphorylation levels of the regulatory proteins involved in autophagy and endothelial-mesenchymal transition (EndMT). The results showed that G. acuta alleviated cardiac dysfunction and remodelling. The elevated levels of myocardial hypertrophy and fibrosis markers, induced by TAC, decreased significantly after G. acuta intervention. G. acuta decreased the expression of LC3 II and Beclin1, and increased p62 expression. G. acuta upregulated the expression of CD31 and vascular endothelial-cadherin, and prevented the expression of α-smooth muscle actin and vimentin. Furthermore, G. acuta inhibited the PI3K/Akt/FOXO1/3a pathway and activated the Notch signalling. These findings demonstrated that G. acuta has cardioprotective effects, such as alleviating myocardial fibrosis, inhibiting hypertrophy, reducing autophagy, and blocking EndMT by regulating the PI3K/Akt/FOXO1/3a and Notch signalling pathways.

Immunosuppressive gentianellane-type sesterterpenoids from the traditional Uighur medicine Gentianella turkestanorum.

Whole plants of Gentianella turkestanorum are commonly used as a traditional Uighur medicine. A phytochemical investigation led to the isolation of eight undescribed gentianellane-type sesterterpenoids (18-epi-nitidasin, gentianelloids D-F, and 18-epi-gentianelloids C-F), one undescribed 11,12-seco-gentianellane (18-epi-alborosin), and three known analogs (nitidasin, gentianelloid C and alborosin) among which gentianelloid C was found for the first time from a natural source. The structures of these compounds were elucidated by extensive spectroscopic analyses (including 1D and 2D NMR, HRMS, IR, and specific rotation) and in the case of 18-epi-gentianelloid C by the single-crystal X-ray diffraction analysis. A putative biosynthetic route for these sesterterpenoids was proposed. The immunosuppressive activity of the isolated compounds was also evaluated by their ability to inhibit the proliferation of T cells and T cell cytokine IFN-γ production. Nitidasin suppressed IFN-γ production with an IC50 value of 16.50 µM, while gentianelloid F and alborosin inhibited the proliferation of and IFN-γ production in T cells with IC50 values of 12.40-14.66 µM.

Gentianella acuta prevents acute myocardial infarction induced by isoproterenol in rats via inhibition of galectin-3/TLR4/MyD88/NF-кB inflammatory signalling.

Gentianella acuta (G. acuta), as a folk medicine, was used to treat heart disease by the Ewenki people in Inner Mongolia. However, the effect of G. acuta on acute myocardial infarction (AMI) is not clear. To explore the mechanisms of G. acuta on isoproterenol (ISO)-induced AMI, rats were administered G. acuta for 28 days, then injected intraperitoneally with ISO (85 mg/kg) on days 29 and 30. An electrocardiogram helped to evaluate the myocardial injury. Serum lactate dehydrogenase (LDH), creatinine kinase (CK) and aspartate aminotransferase (AST) levels were evaluated, and haematoxylin eosin, Masson's trichrome staining and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) staining were used to detect myocardial histological changes. Radioimmunoassay was used to measure serum tumour necrosis factor alpha (TNFα) and interleukin (IL)-6. An enzyme-linked immunosorbent assay kit was used to analyse serum galectin-3 (Gal-3) levels. Immunohistochemistry, Western blotting and reverse transcription polymerase chain reaction were used to examine relevant molecular events. The results revealed that pre-treatment with G. acuta decreased the elevation in the ST segment; reduced serum LDH, CK and AST levels; alleviated cardiac structure disorder; and reduced inflammatory infiltration, abnormal collagen deposition and cardiomyocyte apoptosis that were induced by ISO. Furthermore, pre-treatment with G. acuta inhibited serum Gal-3 levels and Gal-3 expression in heart tissue, and also impeded TLR4/MyD88/NF-кB signalling activation, which ultimately prevented the expression of inflammatory cytokines. The study indicated that pre-treatment with G. acuta protects against ISO-induced AMI, and the protective role may be related to inhibiting Gal-3/TLR4/MyD88/NF-кB inflammatory signalling.

Research on the Potential Mechanism of Gentiopicroside Against Gastric Cancer Based on Network Pharmacology.

BACKGROUND: Gastric cancer was still one of the commonly diagnosed cancer types and the third-most common cause of cancer-related death in the world. Gentiopicroside, which is extracted from the Gentianella acuta, is commonly used in both traditional treatment and modern clinical care; therefore, its anticancer effects have been attracted more attention. However, the systematic analysis of action mechanism of Gentiopicroside on gastric cancer (GC) has not yet been carried out. AIM: A network pharmacology-based strategy combined with molecular docking studies and in vitro validation was employed to investigate potential targets and molecular mechanism of Gentiopicroside against GC. MATERIALS AND METHODS: Potential targets of Gentiopicroside, as well as related genes of GC, were acquired from public databases. Potential targets, and signaling pathways were determined through bioinformatic analysis, including protein-protein interaction (PPI), the Gene Ontology (GO), and the Kyoto Encyclopedia of Genes and Genomes (KEGG). Subsequently, molecular docking and cell experiments were performed to further verify the above findings. RESULTS: Our findings revealed that the anticancer activity of Gentiopicroside potentially involves 53 putative identified target genes. In addition, GO, KEGG, and network analyses revealed that these targets were associated with cell proliferation, metabolic process, and other physiological processes. Furthermore, we have proved that critical compound affected the expression of CCND1, CCNE1, p-AKT and p-P38 at protein levels. These findings provide an overview of the anticancer action of Gentiopicroside from a network perspective; meanwhile, it might also set an example for future studies of other materials used in traditional Chinese medicine (TCM). CONCLUSION: This study comprehensively illuminated the potential targets and molecular mechanism of Gentiopicroside against GC. It also provided a promising approach to uncover the scientific basis and therapeutic mechanism of TCM treating for disease.

Gentianelloids A and B: Immunosuppressive 10,11-seco-Gentianellane Sesterterpenoids from the Traditional Uighur Medicine Gentianella turkestanorum.

Two sesterterpenoids, possessing an unusual 10,11-seco-gentianellane skeleton, gentianelloids A and B, were isolated from a traditional Uighur medicine Gentianella turkestanorum. Through extensive spectroscopic analysis and single-crystal X-ray diffraction, their structures including absolute configurations were unambiguously determined. A plausible biosynthetic pathway for the two compounds was proposed. Both compounds showed remarkable immunosuppressive activity, including inhibition of the proliferation, activation, and cytokine IFN-γ production of T cells. The findings suggested that sesterterpenoids could contribute positively to the therapeutic effects of this popular traditional Uighur medicine.

Diversity of endophytic plant-growth microorganisms from Gentianella weberbaueri and Valeriana pycnantha, highland Peruvian medicinal plants.

Microbial diversity in Peruvian mountain areas is poorly know, specially endophytic microorganisms of medicinal native plants from the Cordillera Blanca. So, nine bacterial and six fungal species were isolated from Gentianella weberbaueri and Valeriana pycnantha. According to 16S rDNA analysis, bacterial strains belong to genera Rahnella, Pseudomonas, Serratia, Rouxiella, and Bacillus; while ITS analysis showed that fungi belong to Pyrenochaeta, Scleroconidioma, Cryptococcus, and Plenodomus genera. Rahnella sp. GT24B and P. trivialis VT20B solubilized tricalcium phosphate and produced siderophores at 10 and 24 °C. Five bacteria strains produced indol-3-acetic acid (IAA) at 10 and 24 °C, where Rahnella sp. VT19B showed more production at 10 °C than 24 °C. Rahnella sp. GT24B, Serratia sp. VT28B, and Rahnella sp. GT25B inhibited Fusarium oxysporum growth up to 100, 78 and 74 %, respectively. R. inusitata VT25B and B. licheniformis GT10B showed high cellulolytic and proteolytic activities. On the other hand, only a few fungi moderately inhibited growth of F. oxysporum, and produced siderophores and cellulases. Most of bacteria inoculated on Medicago sativa "alfalfa" and Triticum aestivum "wheat" seeds got better root development, especially Rahnella sp. GT24B, Rouxiella sp.VT24B, Serratia sp. VT28B, and Rahnella sp. VT34B. Finally, this study is the first report of endophytic microorganisms associated to wild medicinal high-mountain Peruvian plants and it show a valuable microbial diversity and its possible role in promoting growth of crops and wild medicinal plants.

The aqueous extract of Gentianella acuta improves isoproterenol­induced myocardial fibrosis via inhibition of the TGF­ß1/Smads signaling pathway.

Gentianella acuta (G. acuta) is one of the most commonly used herbs in Chinese Mongolian medicine for the treatment of heart disease. Previously, it was found that G. acuta ameliorated cardiac function and inhibited isoproterenol (ISO)­induced myocardial fibrosis in rats. In this study, the underlying anti­fibrotic mechanism of G. acuta was further elucidated. Histopathological changes in the heart were observed by hematoxylin­eosin, Masson trichrome and wheat germ agglutinin staining. Relevant molecular events were investigated using immunohistochemistry and western blotting. The results revealed that G. acuta caused improvements in myocardial injury and fibrosis. G. acuta also inhibited collagens I and III and α­smooth muscle actin production in heart tissue. G. acuta downregulated the expression of transforming growth factor ß1 (TGF­ß1) and notably inhibited the levels of phosphorylation of TGF­ß receptors I and II. Furthermore, G. acuta caused downregulation of the intracellular mothers against decapentaplegic homolog (Smads)2 and 4 expression and inhibited Smads2 and 3 phosphorylation. The results further demonstrated that the mechanism underlying anti­myocardial fibrosis effects of G. acuta was based upon the suppression of the TGF­ß1/Smads signaling pathway. Therefore, G. acuta may be a potential therapeutic agent for ameliorating myocardial fibrosis.

Can we learn from the ecology of the Bohemian gentian and save another closely related species of Gentianella?

Bohemian gentian (Gentianella praecox subsp. bohemica) is an endemic taxon that occurs on the Czech Massif and together with the Sturmian gentian (Gentianella obtusifolia subsp. sturmiana) are the only autumnal species of Gentianella with large flowers in central Europe. Both species have declined dramatically in both population size and numbers of populations. The Bohemian gentian rescue programme, which recommended appropriate management measures, was adopted in 2011. Here we study the ecology of this species, results of the rescue programme and explore the possibilities of using the experience resulting from this programme for improving the viability of the second species. Long-term monitoring of populations of the Bohemian gentian has shown that regular mowing or grazing together with careful litter removal and gap creation are necessary for its survival in the current climatic conditions. We found some ecological differences between these two closely related species of Gentianella. However, our empirical experience of the largest population of the Sturmian gentian at a site where it thrives, and general evidence that gaps are crucial for the successful establishment of Gentianella seedlings, indicate that regular mowing or grazing together with careful litter removal and creation of gaps, should also be recommended as in the case of the Bohemian gentian rescue programme. Artificial gaps are especially crucial for successful seedling regeneration in oligotrophic meadows with dense vegetation, where the last Sturmian gentian populations survive.

Potencial antihiperglicémico de Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle en la absorción intestinal de glucosa in vitro en ratas albinas diabéticas / Antihyperglycemic potential of Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle for in vitro intestinal glucose absorption in diabetic albino rats

Objetivo: Evaluar el potencial antihiperglicémico de la infusión de Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle en la absorción intestinal de glucosa in vitro en Rattus norvegicus cepa Holtzman con diabetes inducida químicamente. Materiales y métodos: Estudio de tipo experimental y analítico. Se realizó el tamizaje fitoquímico de la planta. Para el estudio in vitro se utilizaron veinte ratas albinas machos, a las que se les indujo diabetes utilizando aloxano monohidratado (Sigma-Aldrich, Saint Louis, MO, EE. UU.) en dosis de 120 mg/kg p.c, vía intraperitoneal, y luego fueron distribuidas al azar en dos grupos de diez ratas albinas cada uno. Luego de diez días fueron sacrificados para extraerles un segmento intestinal con la técnica del saco intestinal evertido, y medir el transporte y absorción de glucosa. En el grupo control (intestinos evertidos incubados en solución Krebs-Henseleit glucosado) y en el grupo experimental (intestinos incubados en solución Krebs Henseleit glucosado más la infusión de Gentianella gilgiana al 10% p/v), se tomaron muestras basales de los líquidos de incubación, luego otras muestras a los 15, 30, 45 y 60 minutos para cuantificar la glucosa de los compartimentos mucoso y seroso mediante el método enzimático de la glucosa oxidasa. Resultados: Se evidenciaron menores concentraciones de glucosa absorbida en compartimento seroso intestinal del grupo experimental (G. gilgiana) con diferencia estadísticamente significativa respecto al control (p< 0,05). Conclusiones: La infusión de Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle presenta potencial antihiperglicémico significativo que disminuye la absorción intestinal de glucosa in vitro en ratas albinas diabéticas.

Objective: To evaluate the antihyperglycemic potential of the Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle infusion for in vitro intestinal glucose absorption in Rattus norvegicus of the Holtzman strain with chemically-induced diabetes. Materials and methods: The study had an experimental and analytical design. A phytochemical screening of the whole plant was carried out. For the in vitro study, twenty male albino rats were induced with diabetes using alloxan monohydrate (Sigma-Aldrich, Saint Louis, MO, USA) at a dose of 120 mg/kg bw administered by intraperitoneal route, and then randomly distributed into two groups of ten albino rats each. After ten days, they were sacrificed to extract an intestinal segment using the everted intestinal sac technique, and measure the glucose transport and absorption. In the control group (everted intestines incubated in Krebs-Henseleit solution containing glucose) and the experimental group (intestines incubated in Krebs-Henseleit solution containing glucose plus the Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle infusion at 10% w/v), basal samples of fluids were taken from incubation, and then other samples were taken at 15, 30, 45 and 60 minutes to quantify the glucose from the mucous and serous compartments using the glucose oxidase enzymatic method. Results: Lower concentrations of absorbed glucose were observed in the intestinal serous compartment of the experimental group (G. gilgiana) with a statistically significant difference compared to the control group (p <0.05). Conclusions: The Gentianella gilgiana (Reimers) Fabris ex J.S. Pringle infusion exhibits a significant antihyperglycemic potential by decreasing the in vitro intestinal glucose absorption in diabetic albino rats.

Gentianella turkestanerum Showed Protective Effects on Hepatic Injury by Modulating the Endoplasmic Reticulum Stress and NF-κB Signaling Pathway.

OBJECTIVE: To investigate the protective effects of Gentianella turkestanerum extraction by butanol (designated as GBA) on hepatic cell line L02 injury induced by carbon tetrachloride (CCl4) and hydrogen peroxide (H2O2). METHODS: L02 cells were incubated with 5 µg/mL, 10 µg/mL, 20 µg/mL, 40 µg/mL, 60 µg/mL, 80 µg/mL and 100 µg/mL GBA for 24 hours, and then MTT assay was used to screen the cytotoxicity for GBA. Cells were divided into blank control group, CCl4/H2O2 model group, treated by CCl4 (20 mmol/L) or H2O2 (100 µmol/L); silymarin+CCl4/H2O2 group, treated by CCl4 (20 mmol/L) or H2O2 (100 µmol/L) and 5 µg/mL silymarin; GBA+CCl4/H2O2 group, treated by CCl4 (20 mmol/L) or H2O2 (100 µmol/L) and GBA (5 µg/mL, 10 µg/mL and 20 µg/mL). MTT assay was performed to determine the cellular activity. Malondialdehyde (MDA) content was determined using a commercial kit. The alanine transaminase (ALT), aspartate transaminase (AST) in the supernatant was determined. PE-Annexin V/7-ADD method was utilized to determine the apoptosis of cells. RT-PCR was used to evaluate the expression of endoplasmic reticulum stressrelated genes (CHOP, PERK, IRE1 and ATF6) mRNA. Western blot analysis was performed to determine the expression of CHOP, Caspase 12 and NF-κB protein. RESULTS: Cellular survival after GBA (5 µg/mL, 10 µg/mL and 20 µg/mL) incubation was ≥ 75%. After GBA incubation, levels of ALT and AST showed a significant decrease (P < 0.05), while that of the MDA showed a significant decrease (P < 0.05). The apoptosis in the CCl4 or H2O2 group showed a significant increase compared to the control group (P < 0.05). In contrast, GBA-preincubation could attenuate the cellular apoptosis compared to the CCl4 or H2O2 group, which displayed a dose-dependent manner (P < 0.05). The expression of CHOP, PERK, IRE1 and ATF6 mRNA was significantly up-regulated in the presence of CCl4 or H2O2 (P < 0.05). Whereas, GBA induced a significant decrease in these mRNA thereafter (P < 0.05), together with a decrease in CHOP and Caspase 12 proteins (P < 0.05). Besides, it could attenuate the expression of NF-κB p65 in nuclear protein. CONCLUSION: G. turkestanerum could inhibit the lipid peroxidation and increase the antioxidant activity. Also, it could inhibit the cellular apoptosis through down-regulating the transcriptional level of ERS related genes and proteins. This process was associated with the nuclear translocation of NF-κB p65 protein.

Effects of Four Compounds from Gentianella acuta (Michx.) Hulten on Hydrogen Peroxide-Induced Injury in H9c2 Cells.

In previous studies, Gentianella acuta (Michx.) Hulten was reported to contain xanthones, iridoids, terpenoids, and sterols and is mainly used to cure hepatitis, jaundice, fever, headache, and angina pectoris. In this study, we used bioassay guided fractionation to identify compounds from G. acuta and investigated their activity against hydrogen peroxide (H2O2)-induced apoptosis of H9c2 cells using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The levels of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and glutamate-cysteine ligase catalytic (GCLC) expression were assessed using quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was evaluated using western blot. The results showed that all four compounds had protective effects on H9c2 cells. The transcription levels of HO-1 and GCLC significantly increased in H9c2 cells pretreated with norswertianolin (1), swetrianolin (2), demethylbellidifolin (3), and bellidifolin (4). However, compared to the model group, the transcription levels of Nrf2 were not enhanced by pretreatment with compounds 1, 2, and 4. The protein expression levels of HO-1 and GCLC in H9c2 cells were greater than that in the H2O2-treated group, and the expression of Nrf2 was not significantly changed except by swetrianolin treatment; inhibitors can reverse the protective effect by ZnPP (15 µM), BSO (10 µM), and brusatol (10 µM). The results indicated that the four compounds isolated from G. acuta inhibited the oxidative injury induced by H2O2 by activating the Nrf2/ARE pathway in H9c2 cells and provide evidence that G. acuta may be a potential therapeutic agent for the treatment of cardiovascular diseases.

Protective role of Gentianella acuta on isoprenaline induced myocardial fibrosis in rats via inhibition of NF-κB pathway.

Gentianella acuta (Michx.) Hulten (G. acuta) has been widely used in Mongolian medicines for the treatment of cardiovascular diseases in Ewenki and Oroqen, Inner Mongolia autonomous region, China. The aim of this study was to investigate the effects and related mechanism of G. acuta on isoproterenol (ISO)-induced oxidative stress, fibrosis, and myocardial damage in rats. Male Sprague Dawley rats were randomly divided into the normal control group, ISO induced group and ISO+G. acuta treatment group. Rats were administered with ISO subcutaneously (5 mg/kg/day) for 7 days, and were orally administered simultaneously with aqueous extracts of G. acuta for 21 days. This investigation showed G. acuta treatment ameliorated cardiac structural disorder, excessive collagenous fiber accumulation and cardiac malfunction. Compared with the ISO induced model group, G. acuta treatment increased superoxide dismutase (SOD) activities and glutathione (GSH) level, prevented the rise of malondialdehyde (MDA), and decreased hydroxyproline contents in the heart tissues. Moreover, G. acuta reduced the expression of transforming growth factor ß1 (TGF-ß1) and connective tissue growth factor (CTGF), and inhibited the expression and activation of NF-κB-P65 in myocardial tissues. These results suggested that G. acuta protects against ISO-induced cardiac malfunction probably by preventing oxidative stress, and fibrosis, and the mechanism might be through inhibiting NF-κB pathway.

Growing competitive or tolerant? Significance of apical dominance in the overcompensating herb Gentianella campestris.

As a compensatory response to herbivory, plants may branch vigorously when the growth of dormant meristems is triggered by shoot damage. Undamaged plants, on the other hand, often restrain branching, and this limitation on growth can be considered a cost of tolerance to herbivory. Restrained branching is caused by apical dominance and may, alternatively, be associated with fitness benefits in competitive environments that favor fast vertical growth. To test these hypotheses regarding selection for restrained branching, we compared the performance of two subspecies of the biennial grassland herb Gentianella campestris; the tall, apically dominant ssp. campestris and the short, multi-stemmed ssp. islandica, which shows reduced apical dominance. For both subspecies, we manipulated the height of surrounding vegetation (competition) and damage intensity in grasslands of differing productivity (high, medium, low), and examined population growth rates using matrix population models combined with life table response experiments. In the absence of damage, ssp. campestris exhibited a higher population growth rate than ssp. islandica in the tallest vegetation, however with the growth rate still being below one. In the medium and low productivity environments where the vegetation was shorter, the population growth rate of ssp. islandica was considerably higher than that of ssp. campestris as long as no more than about 50% of the plants were damaged. When plants were damaged, the apically dominant ssp. campestris showed a positive population growth rate (λ > 1) and often overcompensatory seed production in all productivity levels, while ssp. islandica showed no compensation and therefore the population was predicted to decline (λ < 1). We conclude that restrained branching in Gentianella cannot be selected for by competition alone, but that episodes of apical damage are required to maintain the trait. Furthermore, because of the costs of restrained branching, apical dominance should be selected against in grasslands where competition and disturbance are low.

Xanthones isolated from Gentianella acuta and their protective effects against H2O2-induced myocardial cell injury.

In the present study, two new xanthones, (5'S,8'S)-1,3,5,8-tetrahydroxyxanthone(7→2')-1,3,5,8-tetrahydroxy-5',6',7',8'-tetrahydroxanthone (1), 5-hydroxy-3,4,6-trimethoxyxanthone-1-O-ß-D-glucopyranoside (2), and eight known xanthones (3-10) were isolated from the whole plants of Gentianella acuta. Their structures were identified by the spectroscopic analyses (HR-ESI-MS, and 1D and 2D NMR). Meanwhile, cell-protective effects against H2O2-induced H9c2 cardiomyocyte injury and cytotoxic activities of compounds 1-10 were also determined.

Bioactive Constituents from the Whole Plants of Gentianella acuta (Michx.) Hulten.

As a Mongolian native medicine and Ewenki folk medicinal plant, Gentianella acuta has been widely used for the treatment of diarrhea, hepatitis, arrhythmia, and coronary heart disease. In the course of investigating efficacy compounds to treat diarrhea using a mouse isolated intestine tissue model, we found 70% EtOH extract of G. acuta whole plants had an inhibitory effect on intestine contraction tension. Here, nineteen constituents, including five new compounds, named as gentiiridosides A (1), B (2), gentilignanoside A (3), (1R)-2,2,3-trimethyl-4-hydroxymethylcyclopent-3-ene-1-methyl-O-ß-d-glucopyranoside (4), and (3Z)-3-hexene-1,5-diol 1-O-α-l-arabinopyranosyl(1→6)-ß-d-glucopyranoside (5) were obtained from it. The structures of them were elucidated by chemical and spectroscopic methods. Furthermore, the inhibitory effects on motility of mouse isolated intestine tissue of the above mentioned compounds and other thirteen iridoid- and secoiridoid-type monoterpenes (7-10, 13-16, 18, 19, 21, 22, and 25) previously obtained in the plant were analyzed. As results, new compound 5, some secoiridoid-type monoterpenes 7, 10, 12-14, 16, and 17, as well as 7-O-9'-type lignans 31 and 32 displayed significant inhibitory effect on contraction tension at 40 µM.

Cardioprotective effect of the xanthones from Gentianella acuta against myocardial ischemia/reperfusion injury in isolated rat heart.

Gentianella acuta (Michx.) Hulten is widely used for the treatment of arrhythmia and coronary heart disease in Ewenki Folk Medicinal Plants and Mongolian Medicine, popularly known as "Wenxincao" in China. To investigate the potential protective role of the xanthones from G. acuta against myocardial I/R injury in isolated rat heart and its possible related mechanism. The protective role of xanthones on myocardial I/R injury was studied on Langendorff apparatus. The hemodynamic parameters including the left ventricular developed pressure (LVDP), the maximum rate of up/down left intraventricular pressure (±dp/dtmax), coronary flow (CF) and heart rate (HR) were recorded during the perfusion. The results demonstrated that the xanthones from G. acuta treatment significantly improved myocardial function (LVDP, ±dp/dtmax and CF), increased the levels of superoxide dismutase (SOD) and catalase (CAT), succinate dehydrogenase (SDH), malate dehydrogenase (MDH), ATP and the ratio of glutathione and glutathione disulfide (GSH/GSSG), whereas suppressed the levels of Lactate dehydrogenase (LDH), creatine kinase (CK) and malondialdehyde (MDA). Furthermore, the xanthones upregulate the level of Bcl-2 protein and downregulate the level of Bax protein. These results indicated that xanthones from G. acuta exhibited cardioprotective effects on myocardial I/R injury through its activities of anti-oxidative effect and anti-apoptosis effect.

Chemical Composition, Antioxidant and Anticholinesterase Activities of Gentianella azurea from Russian Federation.

Phytochemical study of Gentianella azurea (Bunge) Holub (Gentianaceae) collected in Buryatia Republic (Russian Federation) resulted in the isolation of twenty-one compounds including bellidifolin, bellidin, isobellidifolin, norswertianolin, isobellidifolin-8-O-ß-D-glucopyranoside, orientin, cynaroside, .cosmosiin, apigenin, 4'-O-caffeoylswertiamarin, swertiamarin-6'-O-ß-D-glucopyranoside and sweroside, firstly detected in this species. The extracts and individual compounds were shown to possess antioxidant and anticholinesterase potential.

Efecto hepatoprotector del extracto acuoso de Gentianella nitida en un modelo experimental inducido por paracetamol / Hepatoprotective effect of the aqueous extract of Gentianella nitida in an experimental model induced by paracetamol

El objetivo fue evaluar el efecto hepatoprotector del extracto acuoso de Gentianella nitida (hercampuri) en un modelo experimental inducido por paracetamol. Para evaluar el efecto de hepatoprotección del extracto de Gentianella nitida se empleó paracetamol como inductor del daño hepático. Se analizó in vitro la capacidad antioxidante (DPPH, ABTS, FRAP), el contenido de fenoles totales y flavonoides del extracto acuoso de Gentianella nitida. En el modelo in vivo se trabajó con 24 ratas Holtzman hembras de 2 meses, formándose 4 grupos (n= 6): grupo control, grupo paracetamol, grupo silimarina y grupo Gentianella nitida. Al grupo Gentianella nitida se administró una dosis de 200 mg/kg de peso corporal durante 7 días, seguido del paracetamol 300 mg/kg de peso corporal por 4 días más. Se utilizó silimarina 50 mg/kg de peso como estándar de referencia. En el homogenizado de hígado se midió catalasa, superóxido dismutasa, glutatión S- transferasa, glutatión, TBARs, proteínas totales. En el análisis estadístico se aplicó prueba Kruskal-Wallis, y como prueba pos hoc Mann Whitney. Se trabajó con una significancia p < 0,05. La capacidad antioxidante equivalente a ácido ascórbico (AAEAC-DPPH) fue 56 g AA/mg ss y la capacidad antioxidante equivalente a trolox (TEAC-ABTS) fue 87,7 g trolox/mg ss. Expresado en FRAP fue 98,5 g FeSO4/mg ss. Fenoles totales fue 65,8 g EAG/mg ss y de flavonoides, 11,7 g EQ/mg ss. En el modelo in vivo, el grupo Gentianella nitida tuvo resultados significativos en la actividad de SOD y TBARs (aumento; p< 0,05) y en la actividad de glutatión S-transferasa y glutatión (disminución; p < 0,05). El extracto de Gentianella nitida exhibe capacidad antioxidante en correlación con el contenido de fenoles totales, protegiendo la actividad de las enzimas antioxidantes hepáticas frente al daño de las ROS producidas por el paracetamol.