The Study On The Relation Of Hic-5and Hepatolithiasis Liver Tissue Fibrosis

Objective：In this study, we have collected fibrotic liver tissues normal livertissues, Liver tissue samples for HE staining and Masson staining，The degreeof tissue fibrosis were observed under light microscopy；Detection ofReal-time PCR in liver tissue of type I collagen, type Ⅲ collagen, Hic-5,α-SMA, TGF-β1gene mRNAexpression abundan；The expression of Hic-5, α-SMA, TGF-β1in liver tissues detected by immunohistochemistry andWestern-blot; to explore the pathogenesis of Hic-5in hepatic fibrosis.Methods:1、80cases of liver specimens were selected, including60cases of operationresection for intrahepatic bile duct stones in patients with liver tissue,20casesof hemangioma patients tumor operation resection of adjacent normal livertissues.2、 we stained by HE fibrosis on liver fibrosis group. Determination ofhepatic fibrosis collagen deposition area of collagen Masson. Statisticalanalysis of preoperative patients more than12hours of fasting serumbiochemistry and coagulation index.3、Detection of Real-time PCR in livertissue of type I collagen, type Ⅲ collagen, Hic-5, α-SMA, TGF-β1genemRNA expression abundan.4、 the expression of Hic-5,α-SMA, TGF-β1protein in liver tissues was detected by immunohistochemistry andWestern-blot in liver tissue. The results of optical densityimmunohistochemistry using Imagepro-IPP image system, semi quantitative determination of Hic-5, TGF-β1and α-SMA value. The results of Westernblot were scanned into the computer by Bio-Rad gel image analysis system,gel quantitative analysis software Quantity One to analyze the gray valu.5、SPSS17.0statistical software for statistical tools. Measurement data processingmeans and standard deviations(ˉx±s), among the groups were compared withsingle factor analysis of variance or Krusksl-Wallis H rank sum test, multiplesamples were compared using SNK test, count data using chi square test.Correlation analysis using Pearson correlation. The bilateral α=0.05for thestandard test, P<0.05had statistical significance.Results:1、The analysisbetween the normal group and hepatic fibrosis indexes of liver function andcoagulation index: ALT, AST, TBIL, DBIL and IBIL fibrosis group wassignificantly higher than normal group, Statistical analysis showed significantdifferences, P<0.05.The difference of PT, INR, APTT, TT, PBG in normalgroup and hepatic fibrosis group differences are also has no statisticalsignificance (P>0.05).2、 Real-time analysis showed that, among theexpression of Hic-5, α-SMA, TGF-β1, Col I, Col III mRNA: in normalgroup and hepatic fibrosis group, with the degree of liver fibrosis increasedexpression gradually increased, the degree of fibrosis is heavier, strongermRNA expression.3、immunohistochemistry showed that the optical density,Hic-5, α-SMA, TGF-β1values in normal group and hepatic fibrosis groupwere with the degree of liver fibrosis aggravated gradually increased, theoptical density is highest at severe fibrosis in liver tissue. Correlation analysis was performed between normal group and hepatic fibrosis group, R values aregreater than zero;Hic-5, TGF-β1, α-SMA expression was highly positivecorrelation r=0.986, P<0.001(Hic-5and α-SMA); r=0.984, P<0.001(Hic-5andTGF-β1); r=0.961, P<0.001(α-SMA and TGF-β1).4、Western blot analysisshowed that, in the normal group and hepatic fibrosis group，The expressionof Hic-5,α-SMA, TGF-β1is increased with the degree of fibrosis aggravatedgradually increased,The protein expression was the highes in severefibrosis.Conclusion:1、Hic-5and TGF-β1have Participated the developmentprocess of cholestasis induced liver fibrosis.2、In hepatolithiasis patients, withthe severity of hepatic fibrosis,Hic-5， TGF-β1and α-SMA expressionstrengthene.3、In cholestasis induced liver fibrosis, positive expression ofHic-5and TGF-β1related, there may be a regulatory relationship.