ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2017, Vol. 48 ›› Issue (11): 2076-2083.doi: 10.11843/j.issn.0366-6964.2017.11.008

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miR-130b Inhibits the Differentiation of Rabbit Preadipocytes by Targeting PPARγ

HE Hong-bing, CAI Ming-cheng, LIANG Xiao-hu, JIA Xian-bo, LAI Song-jia*   

  1. Institute of Animal Genetics and Breeding, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 611130, China
  • Received:2017-06-06 Online:2017-11-23 Published:2017-11-23

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.

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