中国糖尿病杂志
中國糖尿病雜誌
중국당뇨병잡지
CHINESE JOURNAL OF DIABETES
2011年
4期
293-297
,共5页
臧丽%母义明%吕朝晖%汪保安%窦京涛%陆菊明%潘长玉
臧麗%母義明%呂朝暉%汪保安%竇京濤%陸菊明%潘長玉
장려%모의명%려조휘%왕보안%두경도%륙국명%반장옥
人白血病相关蛋白16%PPAR反应元件%过氧化物酶体增殖物激活体受体%葡萄糖摄取%3T3-L1脂肪细胞
人白血病相關蛋白16%PPAR反應元件%過氧化物酶體增殖物激活體受體%葡萄糖攝取%3T3-L1脂肪細胞
인백혈병상관단백16%PPAR반응원건%과양화물매체증식물격활체수체%포도당섭취%3T3-L1지방세포
LRP16%PPAR% Glucose uptake% 3T3-L1 adipocytes
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.
目的 探討人白血病相關蛋白(IRP)16對3T3-L1脂肪細胞葡萄糖攝取及過氧化物酶體增殖物激活受體(PPAR)γ活性的影響.方法 利用脂質體轉染及慢病毒介導的RNA榦擾技術構建LRP16過錶達、抑製錶達及對照細胞繫.檢測LRP16對細胞葡萄糖攝取的影響.Luciferase法檢測LRP16對PPAR反應元件(PPRE)相對熒光素酶活性的影響.Westem Blot法檢測LRP16對PPARγ及葡萄糖轉運蛋白(GluT)-4錶達的影響.結果 (1)成功構建LRP16過錶達、抑製錶達及對照細胞繫.(2)過錶達LRP16抑製細胞胰島素刺激的葡萄糖攝取;抑製錶達LRP16促進細胞胰島素刺激的葡萄糖攝取.(3)LRP16劑量依賴性的抑製COS-7細胞PPRE的相對熒光素酶活性;(4)LRP16抑製脂肪細胞PPARγ及GluT-4蛋白錶達.結論 LRP16通過下調PPARγ活性抑製3T3-L1脂肪細胞葡萄糖攝取導緻胰島素牴抗.
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.
