摘要:
目的:在整体水平研究促炎细胞因子IL-17在巨细胞病毒肝炎发病机制中的作用.方法:首先建立MCMV全身播散性感染模型,设立正常对照组、MCMV感染对照组、IL-17阻断组和同型抗体对照组.在实验第7天处死小鼠,Westernblot检测各组肝脏组织中IL-17蛋白表达水平,病理评估各组肝组织损伤程度;罗氏生化分析仪检测小鼠血清ALT水平;双抗体夹心ELISA法检测血清中IL-17水平;RT-PCR检测肝脏组织内IL-17R、IFN-γ和IL-10 mRNA的表达.结果:与MCMV感染对照组及同型抗体对照组相比较,IL-17抗体阻断肝脏组织IL-17表达明显下降(P<0.05),病理损伤程度明显减轻,ALT水平显著降低[(146±15)vs (102±11)vs (37±12),P<0.05];血清中IL-17水平降低[(719.76±6.06) vs(722.1 ±4.62) vs(707.53±8.58),P<0.05];IFN-γ[(0.56±0.06)vs (0.55±0.13)vs (0.96±0.2),P<0.05]与IL-10 mRNA[(0.55±0.073)vs(0.51±0.07) vs(0.903±0.18),P<0.05]的表达明显增加,而IL-17R mRNA表达差异无显著统计学意义[(0.81 ±0.16) vs(0.89±0.38)vs (0.87±0.23),P>0.05].结论:促炎因子IL-17高表达参与了巨细胞病毒肝炎的免疫损伤过程,阻断IL-17有助于改善肝功能,减轻肝组织病理损伤.%Objective:To investigate the role of pro-inflammatory cytokine IL-17 involved in the pathogenesis of cytomegalovirus hepatitis in vivo.Methods:First of all,disseminated infection model was established.Then,mice were randomly divided into 4 groups:normal control group,MCMV-infected control group,IL-17 blockade group,and isotype control group.Mice were sacrificed on day 7 after infection.The levels of IL-17 protein were detected by Western blot.Hematoxylin eosin (HE) staining was performed to evaluate the pathologic change of the liver.Serum ALT levels were detected by a Roche DPPI biochemical analyzer.The level of serum IL-17 was measured by double antibody sandwich ELISA.The expressions of mRNA of IL-17R,IFN-γand IL-10 in liver were detected by RT-PCR.Results:Compared with MCMV-infected mice and isotype control,the blockade of IL-17 inhibited the expression of IL-17 protein in liver (P<0.05).The degree of liver damage reduced obviously.The serum ALT was significantly lower [(146±15)vs (102±11)vs (37±12),P<0.05].The level of serum IL-17 was relatively reduced[(719.76±6.06)vs (722.1±4.62) vs (707.53 ±8.58),P<0.05].The expression of IFN-γmRNA [(0.56± 0.06)vs (0.55±0.13)vs (0.96±0.2),P<0.05] and IL-10 mRNA[(0.55±0.073) vs (0.51 ±0.07) vs (0.903 ±0.18),P<0.05] increased significantly,while that of IL-17R did not change apparently[(0.81±0.16)vs (0.89±0.38) vs (0.87±0.23),P>0.05].Conclusion:The increased expression of pro-inflammatory cytokine IL-17 is involved in the pathogenesis of immune injury in cytomegalovirus hepatitis.The blockade of IL-17 is helpful to relieve the liver damage and improve the liver function.