激活HH信号通路抑制猪ADSCs向脂肪细胞的分化

Activation of HH signaling pathway inhibits the adipogenic differentiation of porcine adipose-derived mesenchymal stem cells

为探讨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,应用油红O染色法检测ADSCs成脂分化过程中的形态变化;采用荧光定量PCR及Western Blot的方法检测HH通路Gli1及脂肪细胞分化转录因子的时序表达。通过嘌呤衍生物2,6,9-三元取代嘌呤(Purmorphamine,PM)激活猪ADSCs细胞中HH信号通路,探讨体外激活HH信号通路对猪ADSCs向脂肪细胞分化的作用及其机制。结果显示,在成脂诱导分化的过程中,经5μmol/mL的PM持续处理,细胞中Gli1的mRNA及蛋白表达量增加;猪ADSCs向脂肪细胞分化的数目减少、细胞内甘油三酯含量降低;脂肪细胞分化关键转录因子C/EBPα和PPARγ的表达受到抑制。以上结果表明,激活HH信号通路可通过下调PPARγ和C/EBPα的表达抑制猪ADSCs向脂肪细胞的分化。

To investigate the effects and molecular mechanisms of HH signalingpathway in the adipogenic differentiation of adipose-derived mesenchymal stemcells（ADSCs）,porcine ADSCs was established and the HH signaling pathway was activated with continuous treatment by purmorphamine.The adipogenic differentiation of ADSCs was analyzed by oil red O staining.The expression of Gli1 and adipogenic transcription factors was checked by Real-timePCR and Western blot.The results showed that：HH signaling pathway was activated bycontinuous treatment with 5μmol/mLpurmorphamine（PM）;The expression level of Gli1 mRNA and protein wasincreased;The number of adipocytes and the content of triglyceride were decreased during adipogenic induction differentiation of porcine ADSCs;In addition,CCAAT/enhancer-binding protein-α（C/EBPα）and peroxisome proliferator-activated receptor-γ（PPARγ）,the adipogenic transcription factors,were repressed by PM.The result indicated that the activation of HH signaling pathway inhibits the adipogenic differentiation of porcine ADSCs,and its inhibition may be due to reducing the expression of C/EBPαand PPARγ.