猪ADSCs的成脂分化及HH通路关键基因和脂肪转录因子的时序表达

Adpogenic differerentiation of porcine adipose-derived mesenchymal stem cells and sequential expression of HH signal pathway related genes and adipocyte transcription factors

为了探讨HH信号通路在脂肪细胞分化过程中的作用机制,以猪脂肪间充质干细胞(ADSCs)为材料,经成脂诱导后,利用油红O染色观察脂肪细胞的形态变化,并检测诱导分化不同时期HH信号通路关键基因Smo、Ptc1、Gli1、Gli2以及成脂关键转录因子C/EBPα、PPARγ的表达情况。结果显示,在ADSCs细胞成脂分化过程中,脂肪细胞的数量逐渐增加,脂滴变大;Smo、Gli1、Gli2基因从第4天开始随诱导天数的增加表达量逐渐降低,而Ptc1的变化趋势与之相反;且C/EBPα、PPARγ的表达从第4天开始逐渐增加。结果表明,ADSCs细胞成脂诱导第4天HH信号通路的活性被全面启动,且随诱导天数的增加其活性逐渐降低。

To investigate the effects and mechanisms of HH signaling pathway during adpogenic differeren- tiation, porcine adipose-derived mesenchymal stem cells （ADSCs） were used as materials. The adipogenic differentiation of ADSCs was analyzed using Oil red O staining. The mRNA expression of HH signal pathway related genes （Smo, Ptc1, Gli1, Gli2） and adipogenic transcription factors （PPARγ, C/EBPα） were determined by real-time PCR. The results showed that the number of fat cells gradually increased and lipid droplets became larger during ADSCs adipogenic differentiation. In addition, Smo, Gliland Gli2 genes were highly expressed at day 4, and then decreased to a low plateau after day 16. However, the sequential expression of PPARγ,C/EBPα and Ptcl appeared to be an opposite manner. These results preminarily indicated that the activity of HH signaling pathway started at day 4 of the adipogenic differentiation of porcine ADSCs and gradually reduced with the increase of days.