11β-羟化类固醇脱氢酶1促进3T3-L1前脂肪细胞分化
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国家自然科学基金资助(30570878)


Effects of 11 beta-hydroxysteroid dehydrogenase type 1 on differentiation of 3T3-L1 preadipocyte
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    目的: 应用3T3-L1细胞模型研究11β-羟化类固醇脱氢酶1(11β-HSD1)与前脂肪细胞分化的关系,探讨11β-HSD1在前脂肪细胞分化及肥胖中的作用。方法:构建11β-HSD1-SiRNA 表达质粒pGCsilencerTM H1/TetO1-11β-HSD1并稳定转染3T3-L1细胞,采用油红染色观察脂滴堆积情况,采用Western blot方法检测11β-HSD1在正常3T3-L1细胞分化过程中的表达;并用Real-time PCR检测脂肪细胞分化相关标志基因的变化,阐明11β-HSD1 对前脂肪细胞成脂分化的影响。结果:正常3T3-L1前脂肪细胞在上述三联诱导分化后脂滴堆积随着分化过程逐渐增加;在pGCsilencerTM H1/TetO1-11β-HSD1转染的3T3-L1诱导分化过程中可见脂滴堆积的减少。在3T3-L1前脂肪细胞分化模型中(days 0,2,4,6,8),11β-HSD1蛋白水平是上调的。3T3-L1前脂肪细胞分化模型中GR在分化模型早期(D4及D6)是上调的,但是在分化后期(D8)很快又下降。在正常3T3-L1前脂肪细胞分化早期LPL上调,随着分化后期FAS、PPARγ等明显上升,Pref-1作为脂肪细胞分化抑制因子,在分化早期升高,在分化后期明显下调。在pGCsilencerTM H1/TetO1-11β-HSD1转染后的3T3-L1细胞分化模型中上述有关标志基因发生变化。结论: 11β-HSD1 作为受体前调节剂促进前脂肪细胞分化,与肥胖和胰岛素抵抗相关。

    Abstract:

    Objective:To investigate the relationship between 11 beta-hydroxysteroid dehydrogenase type 1(11beta-HSD1) and preadipocyte differentiation. Methods:The mouse 11beta-HSD1siRNA plasmids were transfected into 3T3-L1 cells for the examination of the effect of targeted 11beta-HSD1 inhibition on differentiation of 3T3-L1 cells. Differentiation was stimulated with 3-isobutyl-1-methyxanthine(MIX), insulin and dexamethasone(DEX). Oil Red O staining was used to observe the lipid accumulation. The genes expression was detected by real-time PCR and Western blot. Results:Lipid accumulation was significantly inhibited in cells transfected with mouse 11beta-HSD1-siRNA compared with untransfected 3T3-L1 cells. Fewer lipid droplets were detected in transfected cells both prior to stimulation and after stimulation with differentiation-inducing reagents. The expression of adipocyte differentiation-associated markers such as lipoprotein lipase(LPL), fatty acid synthetase(FAS) were downregulated in transfected cells. Similarly, the expression of preadipocyte factor-1(Pref-1), an inhibitor of adipocyte differentiation, was downregulated in transfected cells stimulated by MIX, insulin and dexamethasone. Conclusion:11beta-HSD1 can promote preadipocyte differentiation, and may be an important candidate mediator of obesity and obesity-induced insulin resistance.

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刘 云,孙 岩,朱 亭,程 鹏,胡 刚,丁国宪.11β-羟化类固醇脱氢酶1促进3T3-L1前脂肪细胞分化[J].南京医科大学学报(自然科学版),2007,(1):6-10

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  • 收稿日期:2006-08-19
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