叉头框转录因子O亚族1(FoxO1)去磷酸化抑制猪前体脂肪细胞分化  

Dephosphorylation of Forkhead Box Transcription Factor O1 (FoxO1) Suppresses Differentiation of Pig Preadipocytes

作　　者：史新娥[1] 宋子仪[1] 杨浩[1] 鞠大鹏[1] 陈旭[1] 杨公社[1] 

机构地区：[1]西北农林科技大学,动物科技学院,动物脂肪沉积与肌肉发育实验室,杨凌712100

出　　处：《农业生物技术学报》2011年第5期837-842,共6页Journal of Agricultural Biotechnology

基　　金：国家转基因生物新品种培育重大专项基金(No.2009ZX08009-157B);西北农林科技大学动物科学实验教学示范中心资助项目共同资助

摘　　要：叉头框转录因子O亚族1(forkhead box transcription factorO1,FoxO1)在细胞增殖、分化过程中发挥重要的作用。为了阐明其磷酸化水平对猪前体脂肪细胞分化的影响及其分子机理,本研究分离培养猪前体脂肪细胞,用FoxO1的激活剂渥曼青霉素(wortmannin,WM)处理后,采用油红O染色、油红O化学比色法检测猪前体脂肪细胞分化情况,运用qRT-PCR和Western blot方法分析FoxO1及脂肪细胞分化标志基因mRNA和蛋白的表达情况。结果显示,随着脂肪细胞分化,FoxO1 mRNA和蛋白水平以及磷酸化的FoxO1(p-FoxO1)蛋白水平明显升高;WM处理猪前体脂肪细胞后第5天FoxO1蛋白的表达较对照组显著升高(P<0.05),而FoxO1蛋白磷酸化水平显著下降,FoxO1 mRNA表达水平无显著变化(P>0.05);油红O染色结果显示,脂肪细胞分化受到显著抑制(P<0.05);脂肪细胞分化标志基因PPARγ和C/EBPα mRNA表达显著低于对照组(P<0.05)。研究结果提示,WM能降低FoxO1的磷酸化水平,并通过降低PPARγ和C/EBPα mRNA表达来抑制猪前体脂肪细胞分化。FoxO1去磷酸化可以抑制猪前体脂肪细胞分化。Forkhead box transcription factor O1（FoxO1） plays a critical role in cell proliferation and differentiation in mammals. To interpret effects of the phosphorylation of FoxO1（p-FoxO1） on pig preadipocytes adipogenesis and it＇s underlying molecular mechanism, pig preadipocytes were cultured. After treated with wortmannin（WM）, differentiation state of pig preadipocyte was detected by Oil Red O staining and the expression level of FoxO1 and adipocyte differentiation marker gene were analysed by qRT-PCR and Western blot. The results findings showed that as differentiation proceeded, FoxO1 mRNA, protein and p-FoxO1 protein were all increased. After the pig preadipocytes were treated with WM for 5 days, FoxO1 protein was notably increased（P〈0.05）, but p-FoxO1 protein was decreased significantly, meanwhile FoxO1 mRNA was not much changed. Oil red O staining showed that adipocytes differentiation was attenuated and adipocyte markers, such as PPARγ and C/EBPα, were remarkably lowered（P〈0.05）. Taken together, our data demonstrate WM decreases FoxO1 phosphorylation level and suppresses adipogenesis by reducing PPARγ and C/EBPα mRNA. Differentiation of pig preadipocytes is inhibited by dephosphorylation of FoxO1.

关 键 词：叉头框转录因子O亚族1(FoxO1) 渥曼青霉素 前体脂肪细胞 猪 

分 类 号：S188[农业科学—农业基础科学]