橄榄苦苷对胰岛素抵抗HepG2肝细胞胰岛素信号传导的影响  

Effect of Oleuropein on Insulin Signal Transduction in Insulin Resistant HepG2 Hepatocytes

作　　者：杨双[1] 徐继璞[1] YANG Shuang;XU Ji-pu(Suining Central Hospital,Suining 629000,China)

机构地区：[1]遂宁市中心医院

出　　处：《中国实验方剂学杂志》2018年第20期164-168,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：遂宁市科学技术局科研项目(14)

摘　　要：目的：观察橄榄苦苷（oleuropein,OL）对胰岛素抵抗HepG2肝细胞的胰岛素信号传导的影响。方法：常规复苏细胞,于10%FBS＋1%青-链霉素的DMEM（1 g·L^（-1）葡萄糖）培养基中,37℃5%CO_2细胞培养箱中培养,常规细胞培养传至第3代待用;利用1×10^（-6）mol·L^（-1）胰岛素溶液刺激HepG2肝细胞36 h后建立肝细胞胰岛素抵抗模型,分为模型组,OL组（50μmol·L^（-1））,另设正常HepG2肝细胞为正常组,每组设6个复孔;对数生长期细胞,饥饿培养24 h后,诱导建立胰岛素抵抗模型,进行药物干预36 h后,以细胞计数试剂盒-8（CCK-8）检测OL对细胞活性的影响;OL对胰岛素抵抗HepG2肝细胞干预后,实时荧光定量PCR（Real-time PCR）和蛋白免疫印迹法（Western blot）分别检测肝细胞胰岛素受体（InsR）,胰岛素受体底物-1（IRS-1）,葡萄糖转运蛋白-2（GLUT-2）mRNA和蛋白表达水平。结果：与正常组比较,模型组胰岛素抵抗的HepG2肝细胞活性明显降低（P〈0.05）,胰岛素信号分子InsR,IRS-1,GLUT-2 mRNA和蛋白表达明显降低（P〈0.05,P〈0.01）。与模型组比较,OL干预后胰岛素抵抗的HepG2肝细胞活性显著升高（P〈0.01）;胰岛素信号分子InsR,IRS-1,GLUT-2 mRNA和蛋白表达明显升高（P〈0.05,P〈0.01）。结论：OL能够增加胰岛素抵抗肝细胞活性,上调肝细胞胰岛素信号通路中InsR,IRS-1,GLUT-2 mRNA和蛋白的表达,这可能是其改善胰岛素抵抗的机制之一。Objective： This study aimed to explore the effect of oleuropein（ OL） on insulin signal transduction in insulin resistant HepG2 hepatocytes. Method： Routine resuscitation HepG2 cells were cultured in DMEM medium（ 10% FBS ＋ 1% streptomycin）,incubated in 5% CO_2 cell incubator at 37 ℃,and the routine cell culture was spread to third generation for stand-by. The insulin resistant hepatocyte model was established by stimulating HepG2 hepatocytes for 36 h with 1 × 10^（-6） mol·L^（-1） insulin solution. Then the cells were divided into model group,OL group（ 50 μmol·L^（-1））,and normal HepG2 hepatocytes were also used as a normal group. Each group had 6 duplicate holes and three times repeated. After 24 h of starvation culture,the logarithmic growth phase cells were induced to establish insulin resistance model. After drug intervention for 36 h,the effect of OL on cell viability was detected by cell counting kit-8（ CCK-8）. After intervention by OL,Real-time polymerase chain reaction（ Real-time PCR） and Western blot were used to respectively detect mRNA and protein expression levels of insulin receptor（ InsR）,insulin receptor substrate-1（ IRS-1）,and glucose transporter-2（ GLUT-2） in liver cells.Result： As compared with normal group,cell viability was reduced significantly in model group（ P〈0. 05）,and the mRNA and protein expressions of InsR,IRS-1,GLUT-2 were significantly decreased（ P〈0. 05,P〈0. 01）.As compared with model group,the cell viability of HepG2 cells was significantly increased after OL intervention（ P〈0. 01）; the mRNA and protein expressions of InsR,IRS-1,and GLUT-2 were significantly increased（ P〈0. 05,P〈0. 01）. Conclusion： OL can reduce the blood glucose and improve the insulin resistance,and the mechanism may be related to up-regulating the mRNA and protein expressions of InsR,IRS-1,and GLUT-2 in HepG2 cells.

关 键 词：橄榄苦苷 2型糖尿病 胰岛素抵抗 HepG2肝细胞 胰岛素受体 胰岛素受体底物-1 葡萄糖转运蛋白-2 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学]