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ATP敏感钾通道参与大鼠前脂肪细胞增殖和分化

王曜晖, 郑海燕, 秦娜琳, 余上斌, 刘声远

华中科技大学同济医学院病理生理学系.湖北,武汉 430030；华中科技大学同济医学院免疫学系.湖北,武汉 430030

摘要

为了探讨ATP敏感钾通道在前脂肪细胞增殖分化中作用，该实验用逆转录实时定量PCR方法检测大鼠前脂肪细胞和诱导5d获得的脂肪细胞中该通道磺脲类受体2（sulphonylurea receptor2，SUR2）mRNA表达，探讨该通道阻滞剂格列本脲和激动剂二氮嗪对前脂肪细胞中SUR2mRNA表达的影响；MTT检测前脂肪细胞增殖；流式细胞仪检测细胞周期；油红O染色法检测细胞内脂质含量；Image--Pro Plus5.0软件测量细胞直径；逆转录PCR检测过氧化物酶体增殖物激活受体--#gamma#（peroxisome proliferator--activatedreceptor--#gamma# PPAR--#gamma#）mRNA表达。结果显示：前脂肪细胞及诱导5d获得的脂肪细胞均有SUR2mRNA表达，且后者明显高于前者；格列本脲抑制前脂肪细胞SUR2mRNA表达，剂量依赖性地促进前脂肪细胞增殖，增加G_(2)／M+S期细胞百分比，增加细胞脂质含量，使脂肪细胞直径增大，增加PPAR--#gamma# mRNA的表达；二氮嗪在这些方面的作用与格列本脲相反。以上结果提示，ATP敏感钾通道在前脂肪细胞增殖和分化中可能起调节作用，PPAR--#gamma#可能参与这些作用。

关键词： 大鼠; 钾通道; 磺脲类受体; 脂肪细胞; 肥胖症; 细胞增殖; 细胞分化

Involvement of ATP--sensitive potassium channels in proliferation and differentiation of rat preadipocytes

Wang Yaohui, Zheng Haiyan, Qin Nalin, Yu Shangbin, Liu Shengyuan

Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology.Wuhan 430030,Hubei；China

Abstract

This paper was aimed to investigate the effects of ATP-sensitive potassium channels on the proliferation and differentiation of rat preadipocytes. We examined the expression of sulphonylurea receptor 2 (SUR2) mRNA in preadipocytes and adipocytes obtained by inducing for 5 d and the effects of the inhibitor (glibenclamide) and opener (diazoxide) of ATP-sensitive potassium channels on the expression of SUR2 mRNA in preadipocytes by real-time PCR. Preadipocyte proliferation and cell cycle were measured by MTT spectrophotometry and flow cytometer. The content of intracellular lipid was measured by oil red O staining, cell diameter was determined by Image-Pro Plus 5.0 software and the expression of peroxisome proliferator-activated receptor-#gamma#(PPAR-#gamma#) mRNA was estimated by RT-PCR. SUR2 mRNA was expressed in both preadipocytes and adipocytes obtained by inducing for 5 d, and the expression in adipocytes was obviously higher than that in preadipocytes. Glibenclamide inhibited the expression of SUR2 mRNA in preadipocyte, promoted preadipocyte proliferation in a dose-dependent manner, increased the cell percentages in G_(2)/M + S phase,increased lipid content, augmented adipocyte diameter, and promoted the expression of PPAR-#gamma# mRNA. But the actions of diazoxide were contrary to those of glibenclamide. These results suggest that ATP-sensitive potassium channels regulate the proliferation and differentiation of preadipocytes, and PPAR-#gamma# is probably involved in the effect of ATP-sensitive potassium channels.

Key words: Rat;Potassium channels;sulphonylurea receptor;adipocyte;obesity;Cell proliferation;cell differentiation

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引用本文：

王曜晖, 郑海燕, 秦娜琳, 余上斌, 刘声远. ATP敏感钾通道参与大鼠前脂肪细胞增殖和分化[J]. 生理学报 2007; 59 (1): 8-12.

Wang Yaohui, Zheng Haiyan, Qin Nalin, Yu Shangbin, Liu Shengyuan. Involvement of ATP--sensitive potassium channels in proliferation and differentiation of rat preadipocytes. Acta Physiol Sin 2007; 59 (1): 8-12 (in Chinese with English abstract).