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目的探讨人白血病相关蛋白(LRP)16对3T3-L1脂肪细胞葡萄糖摄取及过氧化物酶体增殖物激活受体(PPAR)γ活性的影响。方法利用脂质体转染及慢病毒介导的RNA干扰技术构建LRP16过表达、抑制表达及对照细胞系。检测LRP16对细胞葡萄糖摄取的影响。Luciferase法检测LRP16对PPAR反应元件(PPRE)相对荧光素酶活性的影响。Western Blot法检测LRP16对PPARγ及葡萄糖转运蛋白(GluT)-4表达的影响。结果 (1)成功构建LRP16过表达、抑制表达及对照细胞系。(2)过表达LRP1 6抑制细胞胰岛素刺激的葡萄糖摄取;抑制表达LRP16促进细胞胰岛素刺激的葡萄糖摄取。(3)LRP16剂量依赖性的抑制COS7细胞PPRE的相对荧光素酶活性;(4)LRP16抑制脂肪细胞PPARγ及GluT-4蛋白表达。结论 LRP16通过下调PPARγ活性抑制3T3-L1脂肪细胞葡萄糖摄取导致胰岛素抵抗。
Objective To investigate the effects of human leukemia-associated protein (LRP) 16 on glucose uptake and peroxisome proliferator-activated receptor (PPAR) γ activity in 3T3-L1 adipocytes. Methods Liposome transfection and lentivirus mediated RNA interference technology was used to construct LRP16 overexpression, inhibition and control cell lines. The effect of LRP16 on glucose uptake was examined. The effect of LRP16 on the relative luciferase activity of PPAR response element (PPRE) was detected by Luciferase assay. The effect of LRP16 on the expression of PPARγ and glucose transporter (GluT) -4 was detected by Western Blot. Results (1) Overexpression of LRP16 was successfully constructed, and the expression of LRP16 was inhibited. (2) Overexpression of LRP16 inhibits cellular insulin-stimulated glucose uptake; inhibition of expression of LRP16 promotes insulin-stimulated glucose uptake in cells. (3) LRP16 dose-dependently inhibited the relative luciferase activity of PPRE in COS7 cells; (4) LRP16 inhibited the expression of PPARγ and GluT-4 in adipocytes. Conclusion LRP16 inhibits glucose uptake by 3T3-L1 adipocytes by down-regulating PPARγ activity, leading to insulin resistance.