作　　者： ; ; ; ; ; ; ; ;

机构地区： 华南农业大学动物科学学院

出　　处： 《动物学报》 2007年第1期143-150,共8页

摘　　要： 本实验采用胶原酶消化法分离猪皮下脂肪前体细胞,用含850nmol/L胰岛素和50nmol/L地塞米松的诱导培养液进行诱导,采用油红O提取法测定了细胞中的甘油三酯含量,同时采用实时定量RT-PCR方法检测了细胞分化过程中聚脂相关基因的表达。结果显示:转录因子PPARγ和C/EBPβ在诱导后12h即迅速表达,SREBP-1mRNA表达水平在诱导后12h出现显著下调,随后逐渐升高,96h达到最高水平;脂肪合成相关酶基因GPDH、FAS、ACC和LPL呈现出与SREBP-1相似的表达模式;脂肪酸转运相关基因aP2、FAT、FATP1与VLDLR的表达量随着细胞分化过程的延长而不断增加,并且与细胞内甘油三酯的含量变化高度相关。本实验结果表明,PPARγ、C/EBPβ和SREBP-1可能是调控猪脂肪前体细胞分化的关键转录因子。猪皮下脂肪组织在聚脂过程中,在分化早期可能以脂肪细胞自身合成脂肪酸为主,而后期则主要依赖细胞外脂肪酸的跨膜转运。这些结果可能有助于揭示脂肪细胞的分化调控规律。 Preadipocytes obtained from subcutaneous adipose tissue of 7 day-old piglets （Large-White × Landrace ） by eollagenase-digestion were exposed to DMEM/F12 medium containing 850 nmol/L insulin （Ins） and 50 nmol/L Dexamethasone （Dex） . The concentrations of cell triglyceride were measured by Oil Red O staining. Total RNA was extracted from the cells and real-time quantitative RT-PCR techniques were adopted to analyze mRNA expression of genes involved in adipogenesis. Results showed that： Rapid increases in PPAR γ and C/EBP β mRNA expression were observed after 12 h of induction, while the expression of SREBP-1 mRNA was down-regulated during the first 12 h of induction and then un-regulated gradually till 96 h after induction. The lipogenic-enzyme related genes including GPDH, FAS, ACC and LPL showed expression patterns similar to that of SREBP-1. The expression of genes involved in fatty acid transportation such as aP2, FAT, FATP1 and VLDLR increased gradually during preadipocyte differentiation and were highly related to intracellular triglyceride accumulation. This study provides evidence that PPAR γ, C/EBP β and SREBP-1 are key transcriptional factors of porcine preadipoeyte differentiation. Lipogenic events seem to be active in the early stage of adipoeyte differentiation, while fatty acid transmembrane activities play a more important role in the later stage of differentiation of porcine adipocytes from subcutaneous adipose tissues. These results may be useful in understanding the mechanism involved in adipocyte differentiantion.

关 键 词： 脂肪生成 脂肪前体细胞 基因表达 猪

领　　域： [生物学]