FSTL1基因促进猪前体脂肪细胞分化成脂的机制

    Mechanism of FSTL1 gene promoting porcine preadipocyte differentiation and lipogenesis

    • 摘要:
      目的  构建猪FSTL1基因过表达载体和RNAi载体,探究FSTL1基因对猪前体脂肪细胞分化成脂的作用及机制。
      方法  通过构建FSTL1基因过表达载体和RNAi载体,在猪前体脂肪细胞中过表达、干扰FSTL1基因并诱导其分化,测定细胞中甘油三酯含量以及脂肪细胞分化相关调节基因的mRNA表达量。
      结果  成功克隆得到猪FSTL1基因cDNA序列,其开放阅读框全长924 bp,并将序列提交到GenBank,登录号为KF021281。成功构建猪FSTL1基因的过表达载体pcDNA3.1-FSTL1和RNAi载体pSilencer 4.1-491以及pSilencer 4.1-530。过表达FSTL1基因促进了猪前体脂肪细胞中甘油三酯生成,干扰FSTL1基因抑制了猪前体脂肪细胞中甘油三酯的生成。FSTL1在猪前体脂肪细胞中使PPARγ2AP2基因表达分别上调了73%和113%,使LPLAdiponectinFASN等基因表达分别下调了44%、79%和16%。
      结论  FSTL1基因通过上调PPARγ2AP2基因,下调LPLAdiponectinFASN基因的表达,促进猪前体脂肪细胞的分化,提高猪前体脂肪细胞中甘油三酯的含量。

       

      Abstract:
      Objective  To construct the overexpression vector and RNAi vector for porcine FSTL1 gene, and investigate the role and mechanism of FSTL1 gene on the differentiation of porcine preadipocytes.
      Method  We constructed the overexpression vector and RNAi vectors of FSTL1 gene, transfected the vectors into porcine preadipocytes, and then induced differentiation of preadipocytes. Triglyceride content and the mRNA expression levels of adipocyte differentiation-associated genes were measured.
      Result  The cDNA sequence of porcine FSTL1 gene was successfully cloned, the length of open reading frame was 924 bp, and the sequence has been submitted to GenBank with the login ID of KF021281. The overexpression vector pcDNA3.1-FSTL1 and the RNAi vectors pSilencer 4.1-491 and pSilencer 4.1-530 of the porcine FSTL1 gene were successfully constructed. Overexpressed FSTL1 gene promoted triglyceride content in porcine preadipocytes, whereas triglyceride content was reduced after FSTL1 gene was interfered. FSTL1 up-regulated the mRNA expression level of PPARγ2, AP2 genes by 73% and 113% respectively, and down-regulated LPL, Adiponectin and FASN genes by 44%, 79%, and 16% respectively.
      Conclusion  The FSTL1 gene promotes the differentiation of porcine preadipocytes through up-regulating the mRNA expression of PPARγ2 and AP2 genes and down-regulating LPL, Adiponectin and FASN genes, and increases triglyceride content in porcine preadipocytes.

       

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