摘要
目的:验证2-花生四烯酸甘油引起L02细胞代谢过程中的胰岛素抵抗及其发生机制。方法:用不同浓度2-花生四烯酸甘油处理L02细胞,使用台盼蓝检测处理对细胞存活率的影响;同时观察细胞油红染色以及检测培养上清谷丙转氨酶(ALT)、谷草转氨酶(AST)的改变;用分光光度计法检测胞浆内超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、诱导型一氧化氮合酶(i NOS)的含量;最后使用Western Blot鉴定2-花生四烯酸甘油的作用。结果:当2-花生四烯酸甘油为0.25μM,处理24h,其所致L02细胞胰岛素敏感性下降,且细胞存活无明显差异,其中约有75%胞内有大量脂质沉积。比较2-花生四烯酸甘油联合胰岛素处理组和胰岛素处理组,前者处理后L02细胞胞浆内SOD、GSH显著下降,MDA、i NOS显著升高,其培养上清中ATL、AST含量均升高。随后的WB实验结果显示,2-花生四烯酸甘油处理组较未处理组其胞浆内GLUT4、GSK-3B、e NOS升高。结论:2-花生四烯酸甘油可以通过对L02细胞通路蛋白GLUT4、GSK-3B、e NOS的影响引发胰岛素耐受,进而造成细胞的氧化-还原失衡导致细胞的损伤。
Objective：To study the mechanism of insulin resistance induced by 2-arachidonylglycerol in L02 cells.Methods：The different concentrations of 2-arachidonylglycerol were used during L02 cells culture and the cell survival rate was detected by trypan blue stained.Meanwhile the AST and ALT of culture liquid su- pernatant were measured,and the intracellular SOD,MDA,GSH and iNOS were detected.The expression of GLUT4,GSK-3B and eNOS were determined by Western Blotting method.Results：After treating with 0.25txM 2-arachidonylglycerol for 24 hours,the L02 cells showed decreased insulin sensitivity.Lipid droplets accumulat- ed in about 75% L02 cells.The ceils treated with 2-arachidonylglycerol showed higher AST and ALT in su- pernatant and higher MDA and iNOS in cytoplasm than cells without treatment.The GLUTd,GSK-3B and eNOS expression increased in ceils with 2-arachidonylglycerol treatment.Conelusion：2-arachidonylglycerol can cause insulin resistance that accompanied by the change of GLUT4,GSK-3B and eNOS expression and im- balance oxidation-antioxidant system in the L02 cells.
出处
《中日友好医院学报》
2015年第3期169-172,共4页
Journal of China-Japan Friendship Hospital
基金
武汉大学自主科研青年教师项目
编号2042014kf0152
