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MiR-125a-5p抑制3T3-L1前体脂肪细胞分化
引用本文:纪洪雷,宋成创,李优磊,李岳峰,郑雪莉,杨公社.MiR-125a-5p抑制3T3-L1前体脂肪细胞分化[J].中国生物化学与分子生物学报,2014,30(11):1147-1154.
作者姓名:纪洪雷  宋成创  李优磊  李岳峰  郑雪莉  杨公社
作者单位: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;西北农林科技大学林学院
基金项目:国家自然科学基金项目(No.31101685和No.31072014)~~
摘    要:MicroRNAs(miRNAs)是一类在脂肪组织发育中发挥重要作用的小非编码RNA.为探明miR-125a-5p在3T3-L1前体脂肪细胞中的作用,采用实时qPCR检测了miR-125a-5p在小鼠各组织及3T3-L1前体脂肪细胞分化过程中的表达;使用经化学修饰的miR-125a-5p模拟物agomir及抑制剂antagomir转染3T3-L1前体脂肪细胞,采用实时qPCR和Western印迹检测成脂标志基因Pparγ和aP2的表达,油红O染色观察脂肪细胞脂质积累.结果显示,miR-125-5p在小鼠脂肪组织中高丰度表达,在3T3-L1前体脂肪细胞分化过程中表达下降.过表达miR-125a-5p,与对照组相比,成脂标志基因Pparγ和aP2在mRNA和蛋白质水平均明显下降;油红O染色及定量结果显示脂质积累减少.抑制剂处理结果显示,Pparγ和aP2在mRNA和蛋白质水平均有不同程度上升,但油红O染色及定量结果差异不显著.以上结果表明,miR-125a-5p在脂肪细胞分化中发挥负调控作用.

关 键 词:miR-125a-5p  3T3-L1  脂肪  细胞分化  
收稿时间:2014-03-27

MiR-125a-5p is Implicated in 3T3-L1 Preadipocytes Differentiation
JI Hong-Lei;SONG Cheng-Chuang;LI You-Lei;LI Yue-Feng;ZHENG Xue-Li;YANG Gong-She.MiR-125a-5p is Implicated in 3T3-L1 Preadipocytes Differentiation[J].Chinese Journal of Biochemistry and Molecular Biology,2014,30(11):1147-1154.
Authors:JI Hong-Lei;SONG Cheng-Chuang;LI You-Lei;LI Yue-Feng;ZHENG Xue-Li;YANG Gong-She
Institution:JI Hong-Lei;SONG Cheng-Chuang;LI You-Lei;LI Yue-Feng;ZHENG Xue-Li;YANG Gong-She;Laboratory of Animal Fat Deposition and Muscle Development,College of Animal Science and Technology,Northwest A&F University;College of Forestry,Northwest A&F University;
Abstract:miRNAs are small non coding RNAs that are important for adipose tissue development. To explore the function of miR-125a-5p during the induced differentiation of 3T3-L1 preadipocytes, we analyzed the miR-125a-5p expression levels by real-time qPCR. Following the transfection of miR-125a-5p agomir or antagomir into 3T3-L1 cells, the expression of adipogenic marker genes Pparγ and aP2 were also detected by both real-time qPCR and Western blotting. The lipid droplet formation was assayed by oil red O staining. The results showed that miR-125a-5p was expressed at high levels in adipose tissue, and was downregulated during the process of adipogenesis. The differentiation of 3T3-L1 was impaired with miR-125a-5p overexpression, associated with the decrease of Pparγ and aP2 expression at mRNA and protein levels. The overexpression of miR-125a-5p also led to a notable reduction in lipid droplets as detected by oil red O staining. However, the results of miR-125a-5p antagomir transfection were not able to induce significant cell phenotype changes. In conclusion, we discovered a negative regulatory role of miR-125a-5p in 3T3-L1 preadipocytes differentiation.
Keywords:microRNA-125-5p(miR-125a-5p)  3T3-L1  adipose  cell differentiation  
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