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目的:探讨磷脂酰肌醇3-激酶(PI-3K)通路及其相关蛋白二酰甘油酰基转移酶2(DGAT2)、蛋白激酶C(protein kinaseC,PKC)的亚型PKC-ε在大鼠非酒精性脂肪性肝病(NAFLD)发病中的作用.方法:48只大鼠随机分为6组:正常对照组(N组普通饮食喂养4wk,N2组普通饮食喂养8wk,N3组普通饮食喂养12wk);NAFLD模型组(H1组高脂喂养4wk、H2组高脂喂养8wk、H组高脂喂养12wk);评价各组大鼠肝脂肪变程度、炎症活动和纤维化程度;免疫组织化学法观测各组PKC-ε蛋白表达量;RT-PCR法检测PI-3K(p85a)mRNA和DGAT2mRNA的表达.结果:NAFLD模型组脂肪变性明显,纤维化和炎症活动度记分均显著高于正常对照组H2、H3组脂肪变性分度及纤维化分期比H1组明显增高,气球样变也有显著的差异;H2、H组PKC-ε蛋白表达量与正常对照组比较明显增多(7.68±1.32vs6.68±2.16,8.46±1.19vs5.52±1.05,P<0.05或0.01);PI-3KmRNA(?Ct与正常对照组相比明显升高,且H2、H3组与H1组比较明显升高,PI-3KmRNA表达量在模型组中随脂肪肝程度的加重呈进行性降低DGAT2mRNA(?Ct)与正常对照组比较显著降低,且H2、H3组与H1组比较明显降低,DGAT2mRNA表达量在NAFLD模型组中随脂肪肝程度的加重呈进行性升高.结论:PI-3K通路及其相关蛋白PKC-ε和DGAT2可能与NAFLD发病机制有关.
Objective: To investigate the effects of phosphatidylinositol 3-kinase (PI-3K) pathway and its related proteins diacylglycerol acyltransferase 2 (DGAT2) and PKC isoform PKC in rats Alcoholic fatty liver disease (NAFLD) .Methods: Forty-eight rats were randomly divided into 6 groups: normal control group (normal diet group N for 4 weeks, N2 diet for 8 weeks, N3 normal diet for 12 weeks) ; The rats in NAFLD group were fed with high fat diet for 4 weeks, group H2 was fed with high fat diet for 8 weeks and group H was fed with high fat diet for 12 weeks. The degree of hepatic steatosis, inflammatory activity and fibrosis were evaluated in all groups. Immunohistochemistry (P85a) mRNA and DGAT2 mRNA were detected by RT-PCR.Results: The fatty degeneration of NAFLD model group was significantly higher than that of normal control group (P <0.05), while the score of fibrosis and inflammation activity was significantly higher than that of normal control group The degree of steatosis and fibrosis stage in group H3 were significantly higher than those in group H1, and there were also significant differences in balloon-like changes. The expression of PKC-ε in group H2 and H was significantly higher than that in control group (7.68 ± 1.32 vs 6.68 ± 2.16 , 8.46 ± 1.19vs5.52 ± 1.05, P <0.05 or 0.01). The PI-3K mRNA level was significantly higher than that of the control group The expression of PI-3K mRNA in the model group decreased progressively with the severity of fatty liver (P <0.05), while the expression of DGAT2 mRNA (? Ct) decreased significantly in the three groups compared with the control group And the expression of DGAT2mRNA increased progressively with the increase of fatty liver in NAFLD model group.Conclusion: The PI-3K pathway and its related proteins PKC-ε and DGAT2 may be related to the pathogenesis of NAFLD.