miR-130b靶向PPARγ抑制家兔前体脂肪细胞分化

doi:10.11843/j.issn.0366-6964.2017.11.008

摘要/Abstract

摘要：

旨在探究miR-130b是否可通过靶向PPARγ抑制家兔前体脂肪细胞的分化。采集初生新西兰仔兔肾周脂肪细胞进行原代培养，鸡尾酒法诱导分化后，用RT-qPCR分别测定前体脂肪细胞分化第0、第2、第6、第10天miR-130b表达量及PPARγ（Peroxisome Proliferators-Activated Receptor γ，PPARγ）和C/EBPα（CCAAT/enhancer binding protein α，C/EBPα） mRNA的表达量，生物信息学软件鉴定miR-130b的同源性，预测miR-130b的靶基因；前体脂肪细胞转染miR-130b mimic和miR-130b inhibitor后，检测miR-130b、PPARγ和C/EBPα的表达差异，用油红O染色鉴定前体脂肪细胞分化程度。结果表明：1）在前体脂肪细胞分化过程中，miR-130b、PPARγ和C/EBPα表达量存在差异；分化第8天时，油红染色发现大量橘红色脂滴；生物信息学软件预测PPARγ可作为miR-130b的靶基因，且miR-130b在哺乳动物中具有较高的同源性。2）过表达miR-130b后，mimic组PPARγ和C/EBPα的表达量极显著低于NC组（P < 0.01）。3）抑制表达miR-130b后，inhibitor组PPARγ的表达量极显著高于inhibitor NC组（P < 0.01），inhibitor组C/EBPα表达量显著高于inhibitor NC组（P < 0.05）。4）油红O染色显示分化程度：inhibitor组 > NC组 > mimic组。综上所述，miR-130b在家兔前体脂肪细胞分化过程中差异表达，且可通过靶向PPARγ抑制前体脂肪细胞分化。

Abstract:

The aim of this study was to demonstrate that miR-130b could target PPARγ to inhibit the differentiation of rabbit preadipocytes. Primary perirenal preadipocytes of New Zealand young rabbits were cultured and induced differentiation by cocktail as in vitro experimental model, and then the expression of miR-130b, PPARγ and C/EBPα at 0, 2, 6, 10 d during differentiation were determined by RT-qPCR. Bioinformatics softwares were used to identify homology and predict target genes of miR-130b; The preadipocytes were transfected with miR-130b mimic and miR-130b inhibitor in vitro, and the expression of miR-130b, PPARγ and C/EBPα were detected. The preadipocytes were stained with Oil Red O. The results showed that:1) In the process of preadipocyte differentiation, the expression of miR-130b, PPARγ and C/EBPα were variant; There were a large orange red lipid droplets with Oil Red O staining in the preadipocytes differentiated for 8 days; Bioinformatics software predicted that PPARγ could be used as the target gene of miR-130b, and the sequence of miR-130b had a high homology in mammals. 2) After overexpression of miR-130b, the expression of PPARγ and C/EBPα in mimic group were very significantly lower than that in NC group (P<0.01). 3) After knockdowned miR-130b, the expression of PPARγ in inhibitor group was very significantly higher than that in inhibitor NC group (P<0.01), the expression of C/EBPα in inhibitor group was significantly higher than that in inhibitor NC group (P < 0.05). 4) Oil Red O staining showed that the degree of differentiation:inhibitor group > NC group > mimic group. miR-130b is differentially expressed in the differentiation process of rabbit preadipocytes and may inhibit the differentiation of preadipocytes by targeting PPARγ.

中图分类号:

S829.1

何洪炳, 蔡明成, 梁小虎, 贾先波, 赖松家. miR-130b靶向PPARγ抑制家兔前体脂肪细胞分化[J]. 畜牧兽医学报, 2017, 48(11): 2076-2083.

HE Hong-bing, CAI Ming-cheng, LIANG Xiao-hu, JIA Xian-bo, LAI Song-jia. miR-130b Inhibits the Differentiation of Rabbit Preadipocytes by Targeting PPARγ[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(11): 2076-2083.

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