【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。更多还原

【Abstract】 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 supernatant were measured,and the intracellular SOD,MDA,GSH and i NOS were detected.The expression of GLUT4,GSK-3B and e NOS were determined by Western Blotting method.Results:After treating with 0.25μM2-arachidonylglycerol for 24 hours,the L02 cells showed decreased insulin sensitivity.Lipid droplets accumulated in about 75% L02 cells.The cells treated with 2-arachidonylglycerol showed higher AST and ALT in supernatant and higher MDA and i NOS in cytoplasm than cells without treatment.The GLUT4,GSK-3B and e NOS expression increased in cells with 2-arachidonylglycerol treatment.Conclusion:2-arachidonylglycerol can cause insulin resistance that accompanied by the change of GLUT4,GSK-3B and e NOS expression and imbalance oxidation-antioxidant system in the L02 cells.更多还原