Effect Of HL-10on Stably Transfection With HBV X Gene In HL-7702of Inflammation-related Factors

AIMHepatitis B virus infection (hepatitis B virus, HBV) is one of the major pathogensof viral hepatitis. X gene encoding X protein having a variety of biological functions，including interact with many proteins of the host cell, regulate the expression of thehost cell gene, thereby affecting the host cell, such as signal transduction, cellproliferation and differentiation. The infection of HBV can activate multiple signalingpathways and affect the expression of inflammation-related genes, such as IL-6,cyclooxygenase-2(cyclooxygenase-2, COX-2) and PGE2, etc. This study was toinvestigate the expression of inflammation-related factors in the stably transfected HBVX gene of liver cells, to observe effects of IL-10, in order to further clarify themechanisms of the hepatitis B virus X protein in the pathogenesis of HBV to providesome experimental basis.METHODSThe expression of COX-2protein in HL-7702、HL-7702load、HL-7702X wereexamined by immunohistochemistry technique.Different concentrations ofinterleukin-10(IL-10)(0,40,60,80ng/ml) treated cultured hepatocytes, and learned atdifferent times (24,48h) the expression of COX-2protein. The effect of IL-10on theexpression of COX-2was tested by semi-RT-PCR.Hepatocytes cultured with differentconcentrations of the IL-10(0,40,60,80ng/ml) in vitro,collected the cell supernatant.Detect the expression of supernatant IL-6, TNF-α, PGE2by ELISA.RESULTSThe expression of COX-2protein and mRNA in HL-7702which was stablytransfected HBx compared with the control group and the no-load group wassignificantly higher (P<0.05; P<0.05).There were no significant differences in differenttimes (P>0.05). After joining40,60,80ng/ml of IL-10for24hand48h, the expressionof COX-2protein and mRNA were significantly reduced, and between each group were statistical significances (P<0.05). The expression of IL-6, TNF-α, PGE2were detectedin cell supernatants. The expression of IL-6, TNF-α, PGE2in HL-7702which wasstably transfected with HBx was no significant difference(P>0.05). After adding IL-10in each groups the expression of inflammatory factors had no significant chang.CONCLUSIONThe expression of COX-2in liver cells which was stably transfected with HBxgene was significantly upregulated. Exogenous IL-10can suppress the expression ofCOX-2protein and mRNA, thereby reducing the inflammation of liver cells.There wasno significance in the expression of IL-6, TNF-α, PGE2in cell supernatant,and theinfluence of IL-10was not obvious.