miR-132-3p靶向UCP2调节绵羊前体脂肪细胞分化的研究

doi:10.11843/j.issn.0366-6964.2017.11.006

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (11): 2059-2067.doi: 10.11843/j.issn.0366-6964.2017.11.006

miR-132-3p靶向UCP2调节绵羊前体脂肪细胞分化的研究

师涛¹, 闫晓茹¹, 潘洋洋¹, 景炅婕¹, 乔利英¹, 郭云利¹, 申彩云², 张富权², 刘文忠^1*

1. 山西农业大学动物科技学院, 太谷 030801;
2. 山西省左云县畜牧兽医服务中心, 左云 037100

收稿日期:2017-06-07 出版日期:2017-11-23 发布日期:2017-11-23
通讯作者: 刘文忠,教授,E-mail:tglwzyc@163.com
作者简介:师涛(1992-),男,山西临猗人,硕士生,主要从事肉用绵羊的分子遗传育种研究,E-mail:sxnydxshitao@126.com
基金资助:
国家自然科学基金（31372292）；山西省优势肉用家畜高效安全生产协同创新中心项目

Study on the Regulation of miR-132-3p in Ovine Preadipocyte Differentiation by Targeting UCP2

SHI Tao¹, YAN Xiao-ru¹, PAN Yang-yang¹, JING Jiong-jie¹, QIAO Li-ying¹, GUO Yun-li¹, SHEN Cai-yun², ZHANG Fu-quan², LIU Wen-zhong^1*

1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China;
2. Zuoyun Animal Husbandry and Veterinary Service Center, Zuoyun 037100, China

Received:2017-06-07 Online:2017-11-23 Published:2017-11-23

摘要/Abstract

摘要：

旨在研究miR-132-3p对绵羊前体脂肪细胞中UCP2基因的表达调控机制。本研究以绵羊皮下脂肪细胞为研究对象，检测miR-132-3p与UCP2在绵羊前体脂肪细胞中的作用机制；利用生物信息学软件预测miR-132-3p与UCP2的结合位点，并通过双荧光素酶报告试验验证miR-132-3p与UCP2的靶标关系；在绵羊前体脂肪细胞中过表达miR-132-3p，用RT-qPCR、Western blotting技术检测过表达后UCP2 mRNA和蛋白的表达水平；最后，采用RT-qPCR检测绵羊前体脂肪细胞分化过程中UCP2和miR-132-3p的表达。结果表明，miR-132-3p在UCP2 3'-UTR上存在结合位点，并显著下调UCP2 3'-UTR重组双荧光质粒的相对荧光活性（P < 0.05）；过表达miR-132-3p显著下调UCP2 mRNA的表达（P < 0.05），且明显降低UCP2蛋白的表达量；绵羊前体脂肪细胞分化中期miR-132-3p与UCP2的表达存在负相关关系。综上表明，miR-132-3p通过特异性结合UCP2 3'-UTR抑制UCP2在mRNA和蛋白水平的表达，在一定程度上促进绵羊前体脂肪细胞的分化。

Abstract:

This study aimed to explore the regulatory mechanism of miR-132-3p targeting UCP2 expression in ovine preadipocytes. In this study, ovine subcutaneous fat cells were used to investigate the mechanism of miR-132-3p targeting UCP2 in ovine preadipocytes. The binding sites of miR-132-3p to UCP2 were predicted by bioinformatics softwares, and were verified via double luciferase report system. Expression of UCP2 mRNA and protein were detected by RT-qPCR and Western blotting, respectively after overexpressing miR-132-3p. The expression of UCP2 mRNA and miR-132-3p were detected by RT-qPCR during differentiation stage in ovine preadipocyte. The results showed that miR-132-3p had binding site to UCP2 3'-UTR and significantly down-regulated the relative fluorescence activity of recombinant double fluorescent plasmid(P<0.05). Overexpressing miR-132-3p significantly down-regulated both UCP2 mRNA(P<0.05) and protein expressions. At last, we found a negative correlation between the expression of miR-132-3p and UCP2 at the middle stage during ovine preadipocytes differentiation. In conclusion, miR-132-3p inhibits UCP2 expression at both levels of mRNA and protein by specifically binding to UCP2 3'-UTR, and thus promotes ovine preadipocytes differentiation to a certain extent.

中图分类号:

S826.2

师涛, 闫晓茹, 潘洋洋, 景炅婕, 乔利英, 郭云利, 申彩云, 张富权, 刘文忠. miR-132-3p靶向UCP2调节绵羊前体脂肪细胞分化的研究[J]. 畜牧兽医学报, 2017, 48(11): 2059-2067.

SHI Tao, YAN Xiao-ru, PAN Yang-yang, JING Jiong-jie, QIAO Li-ying, GUO Yun-li, SHEN Cai-yun, ZHANG Fu-quan, LIU Wen-zhong. Study on the Regulation of miR-132-3p in Ovine Preadipocyte Differentiation by Targeting UCP2[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(11): 2059-2067.

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miR-132-3p靶向UCP2调节绵羊前体脂肪细胞分化的研究

Study on the Regulation of miR-132-3p in Ovine Preadipocyte Differentiation by Targeting UCP2

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