Effects Of Autoimmune Hepatitis Correlation Factor On Synthesis Of Asialoglycoprotein Receptor In HepG2 Cells

Background:Autoimmune hepatitis is related with body autoimmune,and etiology and pathogenesis have been still not defined.It was reported that two aspaects were involved in the mechanisms of liver damage with autoimmune hepatitis:T cellmediated cytotoxicity and antibody-dependent cell-mediated cytotoxity.,which is the major mechanism has not been confired.But,cytokines changes throughout the damage mechanism.Asialoglycoprotein receptos is currently considered to play an important role in the pathogenesis of autoimmune hepatitis.It was reported that significant changes in ASGPR expressions and function in Hep G2 cells took place by treatment with IL-1、IL-6 and so on.However, the influence of autoimmune hepatitisrelated cytokines on ASGPR is partially unkown. And this research would improved our knowing to the pathogenesis of asialoglycoprotein receptor and damage of autoimmune hepatitis Objective:To research the expression of ASGPR in patients of autoimmune hepatitis and influence of autoimmune hepatitis related cytokines TNF-α,IL–2, and IL–12 on synthetic of ASGPR in Hep G2 cells, so this research could providing new clues to the mechanisms of liver damage of autoimmune hepatitis. Mehhods:1.The expression changes of ASGPR1 m RNA and ASGPR2 m RNA of the Hep G2 cell were detected by meaning of RT-PCR after the cytokines stimulation in 2h, 20 h and 24 h in TNF-α,IL-2 and IL-12.2.The ASGPR protein changes on the surface were detected using Western blot method after 2h,20 h,24h stimulation of TNF-α、IL-2、IL-12 on the Hep G2 cell.3.The expression status of the ASGPR on the hepatocyte surface of AIH patients using immunohistochemical method. Results:1.TNF-α was able to synchronously work on ASGPR1 m RNA and ASGPR2 m RNA with different expression in 2h and 20h(P=0.002,P=0.000);but IL-2 could not so.ASGPR1 m RNA dropped first and then showed in an increase trend.There was a big difference between 20 h and 24h(P=0.046),ASGPR2 m RNA rose first and then fell, and the expression status in 2h- 20 h,and 2h- 24 h were of statistic value(P=0.001、P=0.000); IL-12 could not synchronously work on ASGPR1 m RNA and ASGPR2 m RNA.Compared to the expression in 24 h,that was much higher(p=0.042)in 2h,and ASGPR2 m RNA rose first and then fell,but with no differences among the three groups(χ2=3.289,P=0.193).2.The ASGPR expression on the surface of Hep G2 cell increased under the stimulation of TNF-α(P>0.05), and decreased under the stimulation of IL-2, and also rose at first and then turned to fall with lower expression(P<0.05), under the stimulation of IL-12 in 24h(P<0.05).3.Reduced ASGPR expression was shown in hepatocyte of the AIH patients,and expressed on the surface of bile. Conclusion:1.TNF-α, IL-12 and IL-2 could affect the synthesis of ASGPR directly The surface ASGPR were involved in pathogenesis and damage mechanisms of autoimmune hepatitis.2. TNF-αtraetment caused equal actvation of both ASGPR1 m RNA and ASGPR2 m RNA in 24 h,while IL-2/IL-12 could not.