Based On The TGF-?1/Smad2 Signaling Pathway To Explore The Mechanism Of Guizhi Fuling Pills In Protecting And Treating Hepatic Fibrosis

Objective:1.By using the data mining software of the Traditional Chinese Medicine Inheritance Auxiliary Platform(V2.5),observe and summarize the rules and prescriptions of Chinese Medicine for treating hepatic fibrosis from 2015 to 2020.2.Through animal experiments and cell experiments in vitro,explore the efficacy,characteristic and mechanism of Guizhi Fuling Pills in the treatment hepatic fibrosis.In order to make clear whether Guizhi Fuling Pills inhibit HSCs activation pass through the TGF-?1/Smad2 signaling pathway and inhibit hepatic sinusoid capillarization to analyze whether Guizhi Fuling Pills can prevent and treat hepatic fibrosis.Methods:1.Data mining:Collect the prescriptions of traditional Chinese medicine for hepatic fibrosis from 2015 to 2020 on CNKI and use the Traditional Chinese Medicine Inheritance Auxiliary Platform(V2.5)software to analyze the four qi,five flavors,meridian tropism,and drug prescription rules of high-frequency medication.2.Animal experiment:60 male SD rats were randomly divided into 6 groups after adaptive feeding for 1 week,which were normal control group,model control group,Guizhi Fuling Pill low,medium and high dose group,and Dahuang Zhechong Pills group using carbon tetrachloride-induced hepatic fibrosis model,the treatment group was given low,medium and high doses of Guizhi Fuling Pills and Dahuang Zhechong Pills at the same time,the normal control group and the model control group were given normal saline.During the experiment observe and record the general conditions of the rats,adjust the amount of gavage according to changes in body weight,and sacrifice after 8 weeks of treatment.Collect liver tissue and blood samples,use HE and Masson staining to observe the degree of hepatocyte injury and hepatic fibrosis in rat liver tissues,and test ALT,AST,PIIINP,TGF-?1,pSmad2,a-SMA,vWF,CD31,Col-III.Scanning electron microscopy was used to observe the changes of hepatic sinusoidal endothelial cell aperture and basement membrane.3.Cell experiment:Hepatic stellate cells(HSC-T6)were cultured in vitro,and Guizhi Fuling pills-containing serum were used to interfere with HSC-T6 cells induced by TGF-?1 and observe cell viability,proliferation and migration.After extracting protein and mRNA,observe pSmad2,?-SMA,Col-?,MMP-9,TIMP-1 expression level.Results:1.The traditional Chinese medicine for treating hepatic fibrosis is mainly promoting blood circulation and removing blood stasis,and preferring to use cold,warm and flat drugs in the Four Qi,bitter,dry and pungent in the Five Flavors and belongs to liver,spleen and heart in meridian;2.Results of animal experiments:?Guizhi Fuling Pills can improve the general condition of hepatic fibrosis induced by carbon tetrachloride;?Guizhi Fuling Pills can reduce the infiltration of inflammatory cells and reduce the proliferation of fibrous tissue,liver weight and relative liver index;?Guizhi Fuling Pills can reduce the expression of ATL,AST,P?NP,TGF-?1/Smad2,?-SMA,Col-?,vWF,CD31,maintain the number of fenestrations,and inhibit the formation of continuous subendothelial basement membrane.3.Cell test results:The serum containing Guizhi Fuling Pills could reduce the proliferation of cells induced by TGF-?1,reduce cell proliferation,migration and invasiveness,decrease pSmad2,?-SMA,Col-? mRNA,TIMP-1mRNA and increase MMP-9 mRNA expression.Conclusions:1.In the past 6 years,the treatment of hepatic fibrosis with traditional Chinese medicine is mainly to promote blood circulation and remove blood stasis.It is basically consistent with the prescription of Guizhi Fuling Pills.2.Animal experiment research:Guizhi Fuling Pills can improve the general condition and hepatic injuries of hepatic fibrosis rats and have a certain protective effect on the liver;Guizhi Fuling pill can inhibit the expression of TGF-?1/Smad2 in CCl4 induced liver fibrosis SD rats,thus reducing the activation of HSCs;Guizhi Fuling pill can reduce the deposition of liver extracellular matrix;reducing the degree of capillarity of the hepatic sinusoids.In summary,it may be an important mechanism for Guizhi Fuling Pills in preventing and treating hepatic fibrosis.3.Cell experiment research:The serum containing Guizhi Fuling Pills can inhibit the proliferation and invasion of HSC-T6 induced by TGF-?1 and reduce the expression of TGF-?1/Smad2 signal pathway,inhibit the activation of HSCs,reduce the production of extracellular matrix and promote its degradation,and alleviate the degree of liver fibrosis.