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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 investigate the expression of glucagon receptor (GCGR) in ob / ob mouse liver and hepatocellular carcinoma HepG_2 cells after steatosis. Methods The body weight, liver function, blood glucose, blood lipid and insulin of 10-week-old male ob / ob mice and control mice were detected by fluorescence quantitative PCR, Western blot and immunohistochemistry respectively Difference and localization of gcgr gene and protein in liver of group mice. HepG_2 cells were treated with different concentrations of oleic acid, the degree of adipose tissue was observed by oil red O staining, and the expression of gcgr mRNA and protein was detected by QPCR and Western blot. Results GCGR was widely expressed in many tissues and expressed more in the liver. The ob / ob mice showed obvious fatty liver disease. Immunohistochemistry showed that the obese ob / ob mice were significantly higher in the liver than in the normal control mice (P <0.05). The expression of GCGR was significantly lower than that of the control group (0.761 ± 0.211 vs 1.200 ± 0.346, P <0.05). Compared with the control group, the expression of GCGR was decreased and the mRNA expression level was 0.709 ± 0.174 vs 1.000 ± 0.000 , The difference was statistically significant (P <0.05). The expression of GCGR mRNA and protein in HepG_2 cells treated with different concentrations of oleic acid decreased gradually with the increase of oleic acid concentration (P <0.05). Conclusion Fatty liver lesions reduce the level of hepatic GCGR expression, which may affect the liver glucose and lipid metabolism.