肝脏脂肪变对胰高血糖素受体的表达影响  

Effect of Hepatic Steatosis on the Expression of Glucagon Receptor

作　　者：王倩倩[1] 肖元元[1] 祝超瑜[1] 魏燕燕[1] 李旭[1] 张明亮[1] 韩峻峰[1] 魏丽[1] 贾伟平[1] 

机构地区：[1]上海交通大学附属第六人民医院内分泌代谢科、上海市糖尿病研究所、上海市糖尿病重点实验室,200233

出　　处：《医学研究杂志》2016年第9期33-38,共6页Journal of Medical Research

基　　金：国家自然科学基金资助项目(面上项目)(81370956);上海交通大学无锡研究院交大专项基金资助项目(2011JDZX021)

摘　　要：目的观察胰高血糖素受体(GCGR)在ob/ob小鼠肝组织以及肝癌细胞株HepG_2细胞脂肪变后表达变化。方法分别检测10周龄的雄性ob/ob小鼠和同窝野生对照小鼠的体重、肝功、血糖、血脂以及胰岛素等指标,用荧光定量PCR方法、Western blot法和免疫组化分别检测两组鼠肝脏中gcgr基因、蛋白表达的差异以及定位情况。不同浓度油酸处理HepG_2细胞,油红O染色观察HepG_2细胞脂肪病变程度,QPCR法和Western blot法检测细胞gcgr mRNA和蛋白的表达变化。结果 GCGR广泛表达于多种组织,以肝脏表达较多,ob/ob小鼠呈现明显的脂肪肝性病变,且免疫组化结果发现,ob/ob小鼠与正常对照小鼠相比,肝脏中GCGR表达减少;mRNA表达水平分别为0.709±0.174 vs 1.000±0.000,与对照组比较,差异有统计学意义(P<0.05);蛋白表达水平分别为0.761±0.211 vs 1.200±0.346,与对照组比较,差异有统计学意义(P<0.05)。不同浓度梯度油酸处理HepG_2细胞,其GCGR mRNA和蛋白水平表达随油酸浓度增加而逐渐降低,差异有统计学意义(P<0.05)。结论肝脏脂肪性病变降低了肝脏GCGR表达水平,从而可能影响肝脏的糖脂代谢。Objective To determine the expression of glucagon receptor （GCGR） in ob/ob mice liver and in hepatic steatosis HepG2 cell. Methods We detected several indicators such as weight, liver function, blood glucose, blood lipids and insulin concentra- tions of 10 -week -old male ob/ob mice and wild littermates. The expression of gcgr were measured by quantitative PCR, Western blot, and the localization of GCGR was achieved by immunohistochemistry. After HepG2 ceils were treated with different concentrations of oleic acid, oil red O staining was conducted to detect hepatic steatosis, gcgr mRNA and protein expression were measured by QPCR and Western blot,respectively. Results GCGR widely expressed in many tissues, especially in liver, ob/ob mice manifested fatty liver disease, and gcgr expression decreased in ob/ob mice liver（P 〈 0.05）. The GCGR protein expression levels in ob/ob mice liver were significantly decreased （ P 〈 0.05 ）. As concentrations of oleic acid increased, gcgr mRNA and protein levels decreased, and the difference was statistically significant. Conclusion Hepatic steatosis reduces the liver GCGR expression levels, which may affect glucose and lipid metabolism in the liver.

关 键 词：胰高血糖素受体 非酒精性脂肪肝 糖尿病 ob/ob小鼠HepG2细胞 

分 类 号：R5[医药卫生—内科学]