| In China, chronic hepatitis B is the major cause of hepatic fibrosis and cirrhosis. It is also one of diseases threatening peoples health. But the pathogene-sis of hepatic fibrosis is not clear completely. In this study the interaction between hepatic pathology and sera TGFβ1 FN and ALT was investigated via liver biopsy analysis and detections of TGFβ1 FN and ALT in 68 patients with chronic hepatitis B; FN, FN mRNA as well as FN mediated cAMP in the WB rat live epithelial cells in vitro based on TGFβ1 stimulating were detected by RT - PCR, Western Blotting and immuno - gold electronic microscopy. WB cell apoptosis was observed as well. The purpose is to asses the relationship between TGFβ1 and hepatic fibrosis as well as FN expression and apoptosis in hepatic non - solid cells besides HSC and to supply some experimental data in pathogenesis of liver fibrosis in chronic hepatitis.Materials and methods68 patients with chronic hepatitis B, male 54, female 14, age: 16-6, were involved in this study. The diagnosis based on guideline for prevention and treatment of viral hepatitis by the association of infectious disease and parasitolo-gy and the association of hepatology in September 2000. ALT, TGFB 1 and LN were measured and liver biopsies were done in all patients. The pathology was a-nalysis by HE, reticular fibers and Masson triple stains. The inflammatory activity and fibrosis were defined based on HAI scores. The sera FN and TGFβ1 were detected with ELISA (Sigma ELISA Kit).Study on FN protein expression in WB live epithelial cells stimulating with TGFβ1. FN expression in WB live epithelial cells stimulating with TGFβ1 was studied by cell culture at time point 0, 4hr, 8 hr and 24 hr. The cells were ly-sated with 100ul lysate buffer and then added 4ul PMSF, 1ul Aprotin following by collecting the cells and centrifuging at 10000g for 15 min. The supernatants were stored at - 130C for further study. The protein was separated by 10% SDS - PAGE and detected by Western blotting follow by semi - quantitative analysis with GIS gel picture processor system. And FN mRNA expression was detected by RT - PCR in WB live epithelial cells in vitro stimulating with TGFβ1. In brief collecting the cells at time point 0, 4hr, 8 hr and 24 hr, the total RN A was extracted with TRIzol, phenol and chloroform and washed with 75% alcohol. The semi - quantitative analysis of mRNA was done by GIS gel picture processor system.FN mediated cAMP and cell apoptosis in WB live epithelial cells. The cells were cultured with TGFβ1 and collected at time point 0, 4hr, 8 hr and 24he and then digested with 4ml 2. 5% trypsin and 0. 03% EDTA at ratio 1:1 and then centrifuged at 1300g for lOimmune min, washing with Hanks solution twice. The pellet was fixed with glutaraldehyde for immuno - gold electronic microsco-py-ResultsThe sera TGFβ1, FN and ALT detection as well as correlation between inflammatory activity and fibrosis in patients with chronic hepatitis.The results of sera TGFβ1, FN and ALT level were FN 525. 04 +414.97, TGFβ1 33.24 +25.48, ALT 96.62 + 120.45 respectively. There is a close correlation between sera TGFβ1 level and sera FN and ALT level in patients with chronic hepatitis based on SPSS software analysis, correlative coefficient 0. 338 and 0. 336, P =0.019 and 0.020. And also comparing FN level and hepatic in-flammatory activity, FN level and fibrosis were corrective closely, correlative coefficients were 0.341 and 0.285 respectively, P =0.007 and 0.025. Those results shown that sera TGFp,, FN and ALT level were positive correlation in patients with chronic hepatitis; FN level and hepatic inflammatory activity, FN level and fibrosis were positive correlation in patients with chronic hepatitis too; there is no relationship between TGFβ1 and hepatic inflammatory activity as well as fibrosis. Those results elucidated that though it is the important cause of extracellular matrix (ECM) such as FN over expression, TGFp,level is not in parallel with degree of fibrosis in patients with chron...
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