环孢素A对肝细胞中磷酸化AKT蛋白表达的影响  

Effects of cyclosporine A on the expression of phosphorylated AKT in human hepatocytes in vitro and rat hepatocytes in vivo

作　　者：杨柳[1] 刘晓军[2] 王宏宇[1] 曹永[1] 马永华[1] 徐春[1] 

机构地区：[1]武警总医院内分泌科,北京100039 [2]武警总医院药剂科,北京100039

出　　处：《中国比较医学杂志》2016年第6期18-22,共5页Chinese Journal of Comparative Medicine

基　　金：武警总医院科研项目(编号:WZ20130403)

摘　　要：目的观察环孢素A(cyclosporin A,Cs A)对肝细胞磷酸化AKT(phosphorylated AKT,P-AKT)表达的影响,探讨Cs A导致胰岛素抵抗的机制。方法本研究分两部分:(1)体外研究:用不同浓度Cs A(0.1μmol/L、1μmol/L、5μmol/L)作用于人肝细胞(HL7702细胞)48 h,用Western blot方法检测P-AKT蛋白的表达水平。(2)体内研究(动物实验):建立环孢素A诱发的大鼠糖尿病模型,用免疫组化方法分析P-AKT蛋白在糖尿病大鼠肝细胞中的表达水平。结果 (1)各浓度(0.1μmol/L、1μmol/L、5μmol/L)Cs A组与对照组相比,HL7702肝细胞中的PAKT蛋白的表达呈现逐渐下降趋势,差异有统计学意义(P<0.05、P<0.01、P<0.01)。(2)环孢素A引起大鼠血糖升高,在服用环孢素A的第5个月,大鼠平均血糖为9.28 mmol/L,说明环孢素A诱导的糖尿病大鼠模型建立成功,糖尿病大鼠肝细胞中P-AKT蛋白的表达明显低于服药前和正常对照组(P<0.05)。结论服用治疗剂量的环孢素A导致大鼠血糖升高,环孢素A抑制大鼠肝细胞中P-AKT的表达,环孢素A抑制人肝细胞HL7702中P-AKT的表达,提示影响PI3K/AKT信号通路可能是环孢素A导致胰岛素抵抗的机制之一。Objective To observe the effects of cyclosporin A on the expression of phosphorylated AKT in hepatocytes,and to investigate the mechanism of insulin resistance caused by cyclosporin A. Methods This study included two parts. 1. In vitro experiment： Human hepatocyte HL77022 cell line was cultured at different concentrations of cyclosporin A（ 0.1 μmol/L,1 μmol/L,5 μmol/L） for 48 hours. The expressions of phosphorylated AKT（ P-AKT） in HL77022 cells were measured by Western blot assay. 2. Rat in vivo experiment： SD rats were randomly divided into 2groups. The rats in the control group were administrated with distilled water 1 m L/Kg/d. The rats in the cyclosporine group were administrated with cyclosporine 25 mg/Kg/d. The total experiment time was 5 months. The levels of fasting blood glucose and insulin were tested at the end of the experiment. The insulin resistance index and insulin sensitivity index were calculated. The expression of P-AKT in the rat hepatocytes was measured by immunohistochemistry. Results 1. Each group of the HL7702 cells treated by Cs A（ 0. 1 μmol/L,1 μmol/L,5 μmol/L） showed a significantly decreasedexpression of P-AKT（ P 0. 05,P 0. 01,and P 0. 01）. 2. After 5 months of therapy,the fasting blood glucose level of rats in the cyclosporine group was 9. 28 mmol / L, indicating that cyclosporine-induced diabetic rat models were established. The insulin sensitivity index in the cyclosporine group was lower than that in the control group（ P 0. 05）.The expression of P-AKT in liver in the cyclosporine group was significantly lower than that in the control group（ P 0. 05）. Conclusions Therapeutic dose of cyclosporine has hyperglycemic effects on rats. Cyclosporine can reduce the expression of phosphorylated AKT in hepatic tissue in rats and also decrease the expression of P-AKT in human hepatocyte HL77022 cells,which indicate that cyclosporine may cause insulin resistance by interfering PI3 K / AKT signal transduction pathway.

关 键 词：环孢素A 胰岛素抵抗 移植后糖尿病 AKT P-AKT 

分 类 号：R33[医药卫生—人体生理学]