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目的 了解胰岛素降解酶 (IDE)基因、酶蛋白表达水平与胰岛素敏感性的关系 ,并初步探讨胰岛素降解抑制剂氯喹改善胰岛素敏感性的分子机制。方法 以原代大鼠肝细胞胰岛素抵抗(IR)模型为研究对象 ,分别采用免疫印迹技术和逆转录 聚合酶链反应 (RT PCR)技术检测IR细胞模型的IDE酶蛋白表达 (EIP)及基因表达水平 (EIG) ,同时检测IDE活性和反映细胞胰岛素敏感性的14 C 2 脱氧葡萄糖掺入率及14 C 醋酸盐掺入率 ,并观察氯喹对IR细胞模型上述指标的影响。结果IR细胞模型的EIG及EIP水平显著高于对照细胞 ,并与其IDE活性呈显著正相关 ,与两种掺入率呈显著负相关。同时氯喹可明显抑制IR细胞模型以及对照细胞的EIG及EIP水平 ,而且抑制的程度与氯喹作用时间的长短密切相关。结论 大鼠肝细胞的EIG和EIP与其胰岛素敏感性之间呈负相关关系 ;高浓度胰岛素可刺激大鼠肝细胞的IDE基因mRNA及酶蛋白的表达 ,而氯喹则对其起明显的抑制作用 ;胰岛素和氯喹对大鼠肝细胞EIG的影响可能是通过影响IDE基因转录来实现。
Objective To investigate the relationship between insulin gene expression and insulin sensitivity and insulin sensitivity and to explore the molecular mechanism of chloroquine, an inhibitor of insulin degradation, in improving insulin sensitivity. Methods The insulin resistance (IR) model of primary rat hepatocytes was used as the study object. The expression of IDE enzyme protein (EIP) and gene expression in IR cell model were detected by Western blotting and reverse transcription polymerase chain reaction (RT PCR) (EIG). At the same time, the activity of IDE, the incorporation rate of 14 C 2 deoxyglucose and the incorporation rate of 14 C acetate, which reflect the insulin sensitivity of cells, were measured. The effects of chloroquine on the above indexes of IR cell model were observed. Results The levels of EIG and EIP in IR cell model were significantly higher than those in control cells, and were positively correlated with their IDE activity and negatively correlated with the two kinds of incorporation rate. At the same time, chloroquine significantly inhibited the EIG and EIP levels of IR cell model and control cells, and the degree of inhibition was closely related to the duration of chloroquine. CONCLUSION: EIG and EIP in rat hepatocytes have a negative correlation with insulin sensitivity. High concentrations of insulin can stimulate the expression of IDE gene mRNA and protein in rat hepatocytes, while chloroquine has a significant inhibitory effect on it. The effect of insulin and chloroquine on the EIG of rat hepatocytes may be through the influence of IDE gene transcription.