Effect Of HCPT On The Expression Of Collagen Type Ⅰ、Ⅲ And TGFβ1and TGFβ1/Smad Signaling Pathway In Rats With Hepatic Fibrosis

Objective:To study the role of HCPT on experimental hepatic fibrosis of CCl4-inducedrats，and explore its anti-fibrotic molecular mechanism by investigating the changesof collagen type I and III、TGFβ1、Smad3、Smad7expression in liver.Methods:25male and25famale SD rats were randomly and averagely divided into fivegroups: normal control group(A),CCl4-induced hepatic fibrosis group (B) and threetreatment groups given HCPT(C, D, E). Carbon tetrachloride solution(CCl4dissolvedin peanut oil,4:6, V/V)was given intraperitoneally in every other group rat with2ml/kg twice per week for8wk inducing experimental hepatic fibrosis, except innormal control group rats received intraperitoneal injection of physiological saline atthe same dose and frequency. In the meantime, rats in the treatment groups alsoreceived HCPT via intraperitoneal injection at dosages of0.25,0.5and1.5mg/kgrespectively three times a week. At the end of this study, rats in each group wereanesthetized and sacrificed. Expressions of collagen type I and Ⅲ、TGFβ1weredetected by immunohistochemistry. TGF-β signaling downstream molecules Smad3and Smad7were examinated by real-time quantitative PCR and Western blottingassay.Results:1、HCPT could inhibit the expressions of collagen type I and Ⅲ、TGF-β1proteins in fibrotic liver in rats：The semiquantitative histological scores showed thatthe expressions of and collagen type I and Ⅲ collagen、TGF-β1in groups A, D, Esignificantly reduced when compared with in group B （collagen type I:A-1.464±0.272、 D-4.505±0.307、 E-4.613±0.281vs B-6.504±0.310, F=404.116;collagen type Ⅲ: A-1.464±0.272、D-4.168±0.809、E-4.157±0.598vs B-6.132±0.322，F=139.393; TGFβ1: A-1.334±0.342、 D-5.251±0.257、 E-4.940±0.329vsB-7.853±0.433, F=554.028; all P<0.05）. Among them, the expressions of collagen type I and Ⅲ in group D was similar to in in group E (P<0.05）. And the expressionsof TGFβ1in group C,D,E were positively correlated with HCPT concentrations.2、HCPT could downregulate the expressions of TGFβ1、Smad3mRNA andupregulate the expression of Smad7mRNA in fibrotic liver in rats: Compared with ingroup B, the expressions of TGFβ1、Smad3mRNA were significantly decreased andthe expression of Smad7mRNA was considerably increased in group A; while theexpressions of TGFβ1、Smad3mRNA were gradually lowered and the expression ofSmad7mRNA was gradually increased in groups C, D, E than those in group B withdosage-depentent（.TGFβ1mRNA: A-0.281±0.059、C-0.916±0.059、D-0.799±0.029、E-0.521±0.108vs B-1.219±0.112, F=202.093; Smad3mRNA: A-0.279±0.061、C-0.871±0.110、D-0.633±0.074、E-0.431±0.056vs B-1.219±0.112，F=179.439;Smad7mRNA: A-0.899±0.212、C-1.455±0.231、D-1.980±0.217、E-2.374±0.128vsB-0.498±0.080, F=173.821; all P <0.05)3、HCPT could reduce the expression of Smad3protein and increase theexpression of Smad7protein in fibrotic liver in rats: The expression of Smad3proteinwas markedly decreased in groups A, C, D, E than it in group B, while the expressionof Smad7protein was obviously raised. The expressions in groups C, D, E were in adosage-depentent way.（Smad3: A-0.134±0.330、C-0.678±0.049、D-0.482±0.053、E-0.420±0.041vs B-0.856±0.098, F=210.508; Smad7: A-0.345±0.113、C-0.605±0.048、D-0.797±0.061、 E-0.988±0.180vs B-0.101±0.063, F=105.400; all P<0.05）Conclusion：HCPT can inhibit hepatic fibrosis induced by CCl4in rats, which is probablyassociated with its down-regulation on fibrogenic signal transduction of TGFβ1/Smadpathway.