CAJ | 학술논문

目的探讨人白血病相关蛋白(IRP)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.Westem 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LRP16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COS-7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.
목적 탐토인백혈병상관단백(IRP)16대3T3-L1지방세포포도당섭취급과양화물매체증식물격활수체(PPAR)γ활성적영향.방법 이용지질체전염급만병독개도적RNA간우기술구건LRP16과표체、억제표체급대조세포계.검측LRP16대세포포도당섭취적영향.Luciferase법검측LRP16대PPAR반응원건(PPRE)상대형광소매활성적영향.Westem Blot법검측LRP16대PPARγ급포도당전운단백(GluT)-4표체적영향.결과 (1)성공구건LRP16과표체、억제표체급대조세포계.(2)과표체LRP16억제세포이도소자격적포도당섭취;억제표체LRP16촉진세포이도소자격적포도당섭취.(3)LRP16제량의뢰성적억제COS-7세포PPRE적상대형광소매활성;(4)LRP16억제지방세포PPARγ급GluT-4단백표체.결론 LRP16통과하조PPARγ활성억제3T3-L1지방세포포도당섭취도치이도소저항.
Objective To investigate the effect of LRP16 （leukemia related protein 16） gene on glucose uptake and PPARγ activity in 3T3-L1 adipocytes. Methods lipidosome transfection and lentivirus mediated siRNA technology were used to construct the cell lines. 2-deoxy-［3H］-D-glucose was used to measure glucose uptake. Luciferase was used to measure the relative luciferase activities of PPRE (PPAR-response elements) and Western blot was used to measure the expression of PPARγ and GluT4 protein. Results (1) Cell line was successfully established. (2) Over-expression of LRP16 gene inhibited the glucose uptake, and down-expression of LRP16 sitimulated the glucose uptake when stimulated by insulin. (3) LRP16 gene inhibited the relative luciferase activity of PPRE in a dose dependent manner. (4) LRP16 gene inhibited the expression of PPARγ and GluT-4 protein. Conclusions LRP16 gene inhibits the glucose uptake and leads to insulin resistance through inhibiting the activity of PPARγ in 3T3-L1 adipocytes.