ABSTRACT
Objective:To reveal the effective components， targets and possible mechanisms of Qinggan Huayu granules in the treatment of non-alcoholic fatty liver disease （NAFLD） and liver cancer based on network pharmacology and experimental verification， and to provide a basis for its rational interpretation of treating different diseases with same method for NAFLD and liver cancer. Method:Based on databases of traditional Chinese medicine and disease， the network pharmacology was used to screen main active compounds and potential targets of Qinggan Huayu granules for NAFLD and liver cancer. STRING 11.0 was used to analyze the interaction between potential targets. The core targets were selected from the interaction targets by cytoHubba plug-in. The gene ontology （GO） function and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis were performed on the target by Metascape database. At the same time， <italic>in vitro</italic> experiments were conducted to validate the effect of kaempferol， one of the main active ingredients of Qinggan Huayu granules， on hepatocellular carcinoma cell model and NAFLD cell model. Result:A total of 43 potential targets of Qinggan Huayu granules for for NAFLD and liver cancer were screened， corresponding to 136 active ingredients in 8 herbal medicines. Through enrichment analysis of potential targets， there were 20 biological processes， 13 molecular functions， 9 cellular components and 15 signaling pathways. Qinggan Huayu granules regulated biological behaviors of tumors related to liver cancer and NAFLD （such as apoptosis inhibition and oxidative stress） mainly through kaempferol， quercetin， luteolin and other active ingredients for Caspase-3 （CASP3）， tumor protein p53 （TP53）， vascular endothelial growth factor A （VEGFA） and other hub genes. <italic>In vitro</italic> experiments revealed that kaempferol could inhibit cell proliferation in a dose-dependent manner in hepatocellular carcinoma cell model. And kaempferol could modulate the levels of malondialdehyde （MDA） and glutathione peroxidase （GPx）， which were the molecular markers of oxidative stress of NAFLD cell model. Kaempfero also regulated the expression level of CASP3 in hepatocellular carcinoma cell model and NAFLD cell model. Conclusion:The main mechanism of Qinggan Huayu granules in treating liver cancer and NAFLD with concept of treating different diseases with same method is related to systematic synergy effect of multiple compounds （represented by quercetin， luteolin and kaempferol）， multiple targets （represented by VEGFA， TP53 and CASP3） and multiple signaling pathways （represented by oxidative stress and cell apoptosis）.
ABSTRACT
In order to provide a foundation for the development and application of Ophiocordyceps gracilis and increase the new resources of cordyceps,an asexual Paraisaria dubia was isolated from an O. gracilis fruit body. After 10 days of liquid fermentation,white globular mycelium and clear transparent fermentation were produced. The mycelium was extracted by hot water and precipitated with ethanol to obtain intracellular crude polysaccharide. The protein was deproteinized to obtain deproteinized polysaccharide. The intracellular pure polysaccharide was purified by Sepharose 4 B column chromatography and were analyzed by UV,IR,1 H-NMR,and13 CNMR data,as well as GC and HPLC. The results showed that the intracellular polysaccharide of P. dubia was composed of glucose,galactose and mannose with a molar ratio of 25. 54 ∶2 ∶1. It was a β-configuration glycosylic bond,containing pyranoside. The initial connection of polysaccharide was β(1→2)(1→4)(1→6) connection. This experiment provides a theoretical basis for the development and application of P. dubia.
Subject(s)
Fungal Polysaccharides , Chemistry , Galactose , Glucose , Hypocreales , Chemistry , Mannose , Mycelium , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To promote development and utilization of Ophiocordyceps gracilis in xinjiang and provide basic data for researching and sustainable developing medicine fungus related to O. gracilis.</p><p><b>METHOD</b>A white strain SFYT002 isolated from the sclerotium of O. gracilis in Xinjiang was researched by morphological observation, ITS and 18SrDNA sequencing. The ITS and 18SrDNA sequences of the strain were determined, BLAST was compared with the other sequences of Tolypocladium in GenBank. The phylogenetic trees of ITS and 18SrDNA sequences were analyzed in Tolypocladium. In addition, the filter paper method was used to study the antibacterial effects.</p><p><b>RESULT</b>The main morphological characters of this strain were white cotton-like colonies, phialide with inflated base, drastically sharping with partially bending tips, small and transparent budding spores with being always assemble to spearhead and globular, subglobular or ellipse conidiospores. The phylogenetic trees of ITS and 18SrDNA sequences were constructed and analyzed in Tolypocladium. It was resulted that Tolypocladium was confirmed to be monophyletic, and the strain SFYT002 was the same as the systematic position of others of T. inflatum. Meanwhile, the antibacterial test was performed against the 4 common pathogenic bacteria. It was showed that both fermentation and its extracts of different polar from this strain possessed good anti-bacteria capacities.</p><p><b>CONCLUSION</b>The strain SFYT02 was identified as T. inflatum, and inhibited effectively growth of bacteria.</p>