人白血病相关蛋白16基因对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γ活性的影响

·糖尿病基础研究·

臧丽 母义明 吕朝晖 汪保安 窦京涛 陆菊明 潘长玉

作者单位：100853北京，解放军总医院内分泌科

通讯作者：母义明，E-mail: muyiming@301hospital.com.cn

基金项目：973重大基础研究项目 (2006CB503903);中华医学会临床医学科研专项基金(10020050227)

【摘要】 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。

doi:10.3969/j.issn.1006-6187.2011.04.016

Effects of LRP16 gene on glucose uptake and PPARγ activity in 3T3-L1 adipocytes

ZANG Li, MU Yi-ming, LV Zhao-hui, et al. Department of Endocrinoligy, PLA General Hospital, Beijing 100853, China

Corresponding author: MU Yi-ming, E-mail: muyiming@301hospital.com.cn

【Abstract】 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.

【Key words】LRP16; PPAR; Glucose uptake; 3T3-L1 adipocytes