南京农业大学学报2016，Vol.39Issue(2)：249-254,6.DOI:10.7685/jnau.201505022
miR-27b及其靶基因PPARγ在肌内和皮下脂肪细胞中的差异表达分析
Differential expression of miR-27b and its target gene PPARγ in intramuscular and subcutaneous adipocytes
摘要
Abstract
[Objectives] The aim of this paper was to investigate the molecular mechanism underlying differential lipid deposition between porcine intramuscular and subcutaneous preadipocytes,peroxisome pROliferator-activated recptor gamma (PPARγ) mRNA and protein levels were compared in these two types of adipocytes,the micmRNAs (miRNAs)potentially targeting PPARγ gene were also predicted and verified.[Methods] The differentiation abilities of intramuscular and subcutaneous preadipocytes were assessed via oil red O staining.The mRNA and protein levels of PPARγ were detected by quantitative real-time reverse transcription-polymerase chain reaction(qRT-PCR) and Western blot.The potential miRNAs targeting PPARγ were predicted by bioinformatics prediction,and miRNAs expression levels were detected by qRT-PCR.The target relationship between the candidate miRNAs and PPARγ was analyzed by dual-luciferase reporter assay.[Results] Intramuscular preadipocytes had stronger differentiation ability(P＜0.05).Both mRNA and protein levels of PPARγ were higher in intramuscular adipocytes (P ＜ 0.05).Six miRNAs,miR-130a,miR-130b,miR-128,miR-301,miR-27a and miR-27b,were predicted to potentially target PPARγ.Among the six miRNAs,miR-27a and miR-27b had significantly lower expression levels in intramuscular adipocytes (P＜0.05).However,dual luciferase activity analysis revealed that only miR-27b could suppress PPARγ expression(P＜0.05).[Conclusions] Intramuscular preadipocytes possess stronger differentiation ability than subcutaneous preadipocytes in vitro.We infer that the variance of miR-27b expression results in the diverse levels of PPARγ,and could be the reason for differential fat deposition between intramuscular and subcutaneous adipocytes in pigs.关键词
皮下脂肪/肌内脂肪/过氧化物酶体增殖物激活受体γ(PPARγ)/miR-27bKey words
intramuscular adipocytes/subcutaneous adipocytes/peroxisome proliferator-activated recptor gamma (PPARγ)/miR-27b分类
农业科技引用本文复制引用
孙文星,南文婷,谷淑华,韩海银,李艳,褚维伟,陈杰..miR-27b及其靶基因PPARγ在肌内和皮下脂肪细胞中的差异表达分析[J].南京农业大学学报,2016,39(2):249-254,6.基金项目
国家自然科学基金项目(31272423) （31272423）
