Biotechnology Bulletin ›› 2020, Vol. 36 ›› Issue (8): 111-119.doi: 10.13560/j.cnki.biotech.bull.1985.2020-0072

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Lipid Droplet Formation in the Pre-adipocytes of Leptin-overexpressed Pig

HU Bin-yue1,2,4, HU Yang1,3, CHENG Wen-min1,2,4, ZHAO Su-mei2, ZHAO Hong-Ye1,3,4, WEI Hong-Jiang1,4   

  1. 1. Key Laboratory of Animal Gene Editing and Cell Cloning in Yunnan Province,Kunming 650201;
    2. College of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201;
    3. College of Plant Protection,Yunnan Agricultural University,Kunming 650201;
    4. State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan,Kunming 650201
  • Received:2020-01-19 Online:2020-08-26 Published:2020-08-27

Abstract: This work aims to investigate the effect of endogenous leptin overexpression on lipid droplet formation in pig adipocytes and to provide a theoretical basis for studying the molecular mechanism of Leptin regulating lipid metabolism. The subcutaneous adipocytes of leptin-overexpressed and wild-type pig were isolated and cultured under aseptic condition. Then these cells were induced to be differentiated to produce lipid droplets(LDs). The area of a lipid droplet and the content of the lipid were observed by microscope after Oil Red O and Bodipy staining. Then the expression levels of lipid droplet formation-related genes were determined by Q-PCR. The cells differentiated to form LDs after 4 d of induction. The results from the Oil Red O and Bodipy staining showed that the number of lipid droplets and lipid contents in the leptin-overexpressed porcine adipocytes were significantly lower than those in the wild type(P<0.05). Furthermore,the expression levels of lipogenesis-related genes PPAR γ,SCAP,SREPB1 and PLIN2 in the leptin-overexpressed porcine adipocytes were significantly lower compared to the wild type(P<0.01). Leptin overexpression inhibited the formation of LDs by down-regulating PPARγ,SREPB1,SCAP and PLIN2 genes expression in porcine pre-adipocytes.

Key words: leptin, porcine adipocytes, lipid droplets, lipogenesis and metabolism-related genes