·糖尿病分子生物学与遗传学·

肖新华 赵向 曹仁贤 文格波 刘江华

基金项目：国家自然科学基金资助项目(81070667);湖南省教育厅青年基金(10B092)

作者单位：421001 衡阳，南华大学第一附属医院内分泌科

通讯作者：刘江华，E-mail: jianghua990@hotmail.com

【摘要】目的 探讨Exendin-4对3T3-L1前脂肪细胞的分化及糖脂代谢相关基因mRNA表达的影响。 方法 体外培养3T3-L1前脂肪细胞，在脂肪细胞分化成熟过程中的不同时期分别用Exendin-4等干预，采用油红O染色，观察脂肪细胞分化及脂质积聚情况;采用荧光定量PCR检测脂肪细胞糖脂代谢标志基因GLuT-4、PPARγ、HSL mRNA表达水平。酶法测定脂肪细胞的甘油三酯含量。 结果 分化成熟的脂肪细胞经油红O染色可见细胞质内大片脂滴呈亮红色，而未分化细胞不被油红O染色。在脂肪细胞分化第0天和第6天用Exendin-4干预，脂肪细胞内TG的含量较空白组增加(P<0.01)，GLuT-4、HSL、PPARγ mRNA的表达上调(P<0.01);在脂肪细胞分化的第12天干预，Exendin-4对肪细胞分化及相关基因mRNA的表达与空白组无明显差异。 结论 Exendin-4促进脂肪细胞的分化并上调糖脂代谢相关基因GLuT-4、PPARγ、HSL mRNA表达，可能为Exendin-4抗糖尿病的部分作用机制。

【关键词】 Exendin-4;甘油三酯;分化

doi:10.3969/j.issn.1006-6187.2011.07.003

Effects of exendin-4 treatment on differentiation of 3T3-L1 preadipocyte and on mRNA expressions of glucose and lipid metabolic marker genes

XIAO Xin-hua ,ZHAO Xiang,CAO Ren-xian, et al.Department of Endocrinology,The First Affiliated Hospital of Nanhua University, Hengyang 421001, China

Correrponding author:LIU Jiang-hua, E-mail:jianghua990@hotmail.com

【Abstract】 Objective To investigate the effect of exendin-4 on differentiation of 3T3-L1 preadipocytes and mRNA expression of glucose and lipid metabolic marker genes. Method s3T3-L1 preadipocytes were cultured in vitro and diferentiated into the matured adipocytes.Cell diferentiation and lipid accumulation were determined by oil red O staining.Glucose and lipid metabolism marker genes (GluT-4、PPAR-γ and HSL) mRNA expression were evaluated by quantitative real-time PCR. The triglyceride contents of isopropanol extracts prepared from 3T3-L1 preadipocytes were quantitated with an acetylacetone-based colorimetric kit. Result sMature adipocytes showed large lipid droplets within cytoplasm stained by oil red O. Preadipocyte differentiation were promoted, TG content and the mRNA expressions of GluT-4, HSL and PPAR-γ were increased (P<0.01) when 3T3-L1 preadipocytes were treated with exendin-4 at day 0 and day 6. Compared with control group, the effects of exendin-4 on adipocytes differentiation and mRNA expression of these genes had no significant difference when 3T3-L1 preadipocytes were treated at day 12. Conclusion Exendin-4 promotes the preadipocytes differentiation, increases glucose and lipid metabolism marker genes mRNA expression, which may be some new mechanisms of exendin-4 anti-diabetes action.

【Key words】 Exendin-4; Triglyceride; Differentiation