脂联素信号通路分子在非酒精性脂肪性肝病模型构建不同时期的表达变化  

Dynamic expression of adiponectin signaling pathway molecules in a rat model of non-alcoholic fatty liver disease

作　　者：刘浩[1] 时昭红[1] Hao Liu;Zhao-Hong Shi(Department of Gastroenterology,Wuhan First Municipal Hospital,Wuhan 430022,Hubei Province,China)

机构地区：[1]武汉市第一医院消化内科,湖北省武汉市430022

出　　处：《世界华人消化杂志》2018年第28期1645-1650,共6页World Chinese Journal of Digestology

基　　金：武汉市卫生局临床医学科研资助项目;No.WX11C03~~

摘　　要：目的研究肝脏脂联素信号通路分子在大鼠非酒精性脂肪性肝病(non-alcoholic fatty liver disease, NAFLD)形成过程不同时期的表达变化.方法 SD雄性大鼠24只随机分为4组:正常组N组(6只),模型组M4组(6只),模型组M8组(6只),模型组M12组(6只).正常组给予普通饲料喂养, NAFLD模型组给予高脂饲料喂养.分别于第4周末处死M4组大鼠,第8周末处死M8组大鼠,第12周末处死N组和M12组大鼠.采用HE染色法观察各组大鼠肝组织病理学改变;ELISA检测各组大鼠血清脂联素水平;RT-PCR法检测大鼠肝脏AdipoR2、PPARα的mRNA表达; Western blot蛋白印记法检测各组大鼠肝脏AdipoR2、PPARα及磷酸化AMPK蛋白表达.结果肝脏HE染色显示,第12周末时模型组大鼠可见肝细胞肿胀明显,细胞质内可见大量的脂肪空泡,少量肝细胞发生坏死,提示造模成功. ELISA法结果表明,与正常组相比,模型组大鼠第4周末、第8周末、第12周末血清脂联素水平逐渐降低,每两组之间比较差异有显著性(P<0.05). RT-PCR法结果表明,与正常组相比,模型组大鼠肝脏第4周末、第8周末、第12周末AdipoR2、PPARα的mRNA表达逐渐减弱,每两组之间比较差异有显著性(P<0.05). Western blot蛋白印记法结果显示,与正常组相比,模型组大鼠肝脏AdipoR2、PPARα及磷酸化AMPK蛋白表达在第4周末、第8周末、第12周末逐渐减弱,每两组之间比较差异有显著性(P<0.05).结论脂联素信号通路分子在NAFLD形成过程中表达逐渐减弱.脂联素信号通路活性逐渐降低可能是NAFLD形成的机制之一.AIM To detect the dynamic expression of adiponectin signaling pathway molecules in a non-alcoholic fatty liver disease（NAFLD） model. METHODS Twenty-four male SD rats were randomly divided into four groups： a normal group（N, n = 6） and three model groups（M4, M8, and M12, n = 6 each）. The normal group was given an ordinary diet, and the NAFLD model groups were given a high-fat diet. At the end of the fourth week, rats in group M4 were sacrificed. M8 rats were killed at the end of the eighth week, and rats in groups N and M12 were sacrificed at the end of the twelfth weeks. Tissue samples were collected for histopathological examinations. Serum adiponectin was detected by ELISA. The expression of AdipoR2 mRNA and PPARα mRNA was determined by RT-PCR. Protein expression of AdipoR2, PPARα, and phosphorylated AMPK was examined by Western blot. RESULTS HE staining showed that liver cell swelling was obvious at the end of the twelfth weeks, with a large number of fat vacuoles in the cytoplasm and a small number of necrotic liver cells, which suggested that NAFLD was successfully induced. Compared with the normal group, serum adiponectin in the model group rats gradually decreased from week 4 to weeks 8 and 12. Compared with the normal group, the expression of AdipoR2 mRNA and PPARα mRNA in the liver of the model group rats gradually decreased from week 4 to weeks 8 and 12. Compared with the normal group, the protein expression of adiponectin signaling pathway molecules in the liver of the model group rats gradually decreased from week 4 to weeks 8 and 12. There was a significant difference between each two groups（P〈0.05）.CONCLUSION The protein expression of adiponectin signaling pathway molecules decreases gradually in the formation process of NAFLD. The activity decrease of the adiponectin signaling pathway is possibly one of the mechanisms contributing to NAFLD.

关 键 词：非酒精性脂肪性肝病 脂联素 脂联素受体2 过氧化物酶体增殖因子激活受体α 腺苷酸活化蛋白激酶 

分 类 号：R575.5[医药卫生—消化系统]