| Background and ObjectiveHepatic fibrosis is a pathological change caused by chronic liver damage caused by multiple causes and a necessary stage for the development of chronic liver disease to cirrhosis.It is characterized by excessive abnormal deposition of extracellular matrix in the liver and affecting the function of the liver.During the pathogenesis of liver fibrosis,the activation and proliferation of hepatic stellate cells(HSC)is the core link.Although it is now clear that hepatic fibrosis is a reversible process,modern medicine has not found effective drugs to reverse it yet.Hepatic fibrosis belongs to the category of lump in the abdomen causing distension and pain and hypochondriac pain in traditional Chinese medicine.Various studies have shown that Chinese medicine,including compound Chinese Medicine,Chinese herbal medicine and some ingredients of traditional Chinese medicine,has a good effect on the treatment of liver fibrosis.As dried root and rhizome of Polygonum cuspidatum,Polygonum cuspidatum has a long medicinal history.In modern research,it is considered that the heat-clearing and detoxifying effect of Polygonum cuspidatum is related to the pesticide effect of liver protection and cholagogue.antibiosis and antiviral.However,there is no experimental study to reveal the effect and specific mechanism of Polygonum cuspidatum on hepatic fibrosis.To find out the effective components in the Polygonum cuspidatum,which has the effect of anti hepatic fibrosis,so as to provide the basis for the development of anti hepatic fibrosis drugs,the activated rat hepatic stellate cell(HSC-T6)was used as an in vitro cell model,and the anti hepatic fibrosis effect and active components of Polygonum cuspidatum were studied by combination of cell activity and drug separation.Methods1.Drug separation and identification techniques(1)The decoction of Polygonum cuspidatum was made into extractum,and the extractum was extracted with petroleum ether,ethyl acetate and n-butyl alcohol in turn.Then the polar parts were obtained respectively.(2)The polar parts were separated by solvent system separation and high performance liquid chromatography(HPLC),and the isolated monomers were further purified.(3)Modern spectroscopy was used to identify chemical structures.2.Activity verificationCell model:rat hepatic stellate cell line(HSC-T6).(1)Cell Counting Kit-8(CCK-8)assay was used to test the activation and proliferation of HSC-T6 cells.(2)5-ethynyl-2’-deoxyuridine(EdU)stain were used to assess the proliferation of HSC-T6 cell.(3)Western Blot assay was used to detect the expression of Bcl-2 and Bax.3.Statistical analysisAll data were presented as mean±standard deviation.Statistical analysis was carried by SPSS(version 21).Statistical differences were evaluated by t-test and one-way ANOVA test.Value of P<0.05 was considered to be statistically significant.Results1.The ethyl acetate part of Polygonum cuspidatum had a strong inhibitory effect on the proliferation of HSC-T6 cells,followed by petroleum ether part,and the effect of n-butanol part was weak.2.A large number of light yellow needle like crystals were isolated from the ethyl acetate part of Polygonum cuspidatum,and the structure was identified as physcion.3.Compared with the control group,physcion could inhibit the proliferation of HSC-T6 cells in a concentration-dependent manner.4.In the EdU staining experiment,the number and proportion of the positive cells in the emodin methyl ether group were significantly smaller than those in the control group(P<0.001).5.Western Blot showed that the expression of Bcl-2 was decreased by physcion at 10μmol/L and 20μmol/L(P<0.001),and the level of Bax was increased(P<0.001). |
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