Effects Of Unique Granules On The Expression Of Smad3 And Smad7 In Liver Tissue Of Rats With Liver Fibrosis Induced By Carbon Tetrachloride Combined With High Fat Diet

Objective:The study is based on the research of previous experiments,to investigate mechanism of Duyiwei Granule has significant inhibitory effect through the observation of Smad3、Smad7 expression changes in hepatic fibrosis model rats with liver fibrosis, to provide theoreticalbasisfor clinical application and expand clinical of Duyiwei Granule.Methods:Seventy-eight SD rats were randomized into six groups(each group has thirteen) including normal control group, model group, Colchicine group, Duyiwei Granulehigh-dose group, middle-dose group, low-dose group.The rats in model group and treatment groups were subcutaneously injected with 40%CCL4 oil chloramphenic-ol once per three days for eight weeks.plus high lipid forage to establish hepatic fibrosis model, while the normal control group were with olive oil and standard forage. From the first day the experiment,positive medicine group was given colchicines(0.04 5g/kg)gavage,each treatment group of Duyiwei was given high(2.4g/kg),medium(1.2/ kg) and low dose(0.6g/kg) of particles gavage while model and normal control group were given equivalent normal saline for eight weeks. When the experiment ended,all rats were be executed with femoral aetery,and then to test the Hepatic biopsy was performed by optical microscope. Hyaluronic acid(HA), Laminin(LN), procollagen type Ⅲ(PCⅢ) and collagen type Ⅳ(Ⅳ-C) were detected by radioimmunoassay(RIA) and to test the Smad3and Smad7 protein express and mRNA express in the hepatic tissues were detected by immunohistochemical assay and real time PCR method.Results:1.The sensitive indexes in serum liver fibrosis(HA、LN、PCⅢ、Ⅳ-C):Content in serum of rats in the model group was significantly higher than that in normal control group (P<0.01);Compared with the model group,serumindexes in each dose teaatment groupwere significantlyinduced in rats(P<0.01);Compared with the colchicinegroup,have statisttically significant difference were observed in the Duyiwei Granule (P>0.05).2.Pathological changes of liver tissue:The normal control group ofliver tissue structure intact,cytoplasm rich,portal area was noinflammatory cellinfiltrationandhype-rplasia of fibrous tissue;compared with normal control group,the hepatic lobule structure of the rats in the model groupthe serious damage ofliver tissue,there are a large number ofinflam matorycell infiltration,necrosisof liver cellsand fattydegegener-atin;compared with the model group,the existence of each group of liver small leaf structure,arranged inneat,but only asmall amount ofinflammatory cell infiltration,no obvious fiber hyperplasia,nofibrous septaformation.3.1mmuno-histochemical test Smad3 and Smad7 in hepatic tissues of rat with liver fibrosis:a small amount of Smad3 and Smad7 basic expression were visible in the hepatic tissue stromalcells of normal controlgroup rats;Compared with normal control group,the number of significantly increased Smad3,Smad7 expression in hepatic tissue in model group rats,brown yellow cytoplasmicdistribution;compared with model group,each group of ratsliver tissue Smad3,Smad7slightly expressed in the portal area,positive stainingto a lesser degree.4.Smad3^ Smad7mRNA expression level in hepatic tissues:compared with the control group,the expression of model group rats hepatic tissue Smad3mRNAwassing-nificantly enhanced,Smad7mRNA expression decreased, the difference was statistica-lly significant(p<0.05); compared with the model group,the expression of the dose of Smad3 mRNA medicine groupand colchicine group decreased significantly(P<0.0 5),the expression of Smad7mRNA was increased,there is statistically significant differ-rences(P<0.05).Conclusion:①DDuyiwei Granule can significantly reduce main serum hepatic fiber indexex of (HA,LN,PCⅢ,Ⅳ-C) contents in hepatic fibrosis model rats.②Duyiwei Granule can reduce the pathological change of hepatic tissue,the mechanism may be related to inhibiting the expression of Smad3, there is a close relationship between the up regulation of Smad7 expression.