PPARγ基因对脂肪来源干细胞迁移能力的影响

Effect of PPARγgene on the migration ability of adipose-derived stem cells

目的通过抑制过氧化物酶体增殖物激活受体γ(proliferator-activated receptor gamma,PPARγ)基因,研究其对脂肪来源干细胞(adipose-derived stem cells,ADSCs)迁移能力的影响,探讨可能的信号通路。方法提取人ADSCs进行体外培养,分别用含有不同浓度的PPARγ抑制剂双酚A二缩水甘油醚(bisphenol adiglycidyl ether,BADGE)作用于ADSCs,然后通过划痕实验及Transwell小室实验观察ADSCs的迁移情况,分别用PCR、WB方法检测与迁移相关基因MMP2、MMP9、CXCR4及其蛋白表达情况。结果ADSCs迁移能力在二维和三维空间均下降(P<0.05)。二维空间细胞迁移情况:在72 h,对照组迁移率约63%,25μm/L组约43%,50μm/L组约24%。三维空间:与对照组相比,抑制PPARγ基因,24 h时25μm/L及50μm/L组ADSCs穿过小室的细胞数量较对照组分别降低了14%和28%(P<0.05)。抑制PPARγ基因后MMP2、MMP9、CXCR4的基因及其蛋白表达显著降低(P<0.05)。结论抑制PPARγ基因后ADSCs的迁移能力下降,其抑制作用随抑制剂浓度增加而增加,可能的调控机制是抑制了MMP和CXCR4信号通路的传导。

Objective To investigate the effect of peroxisome proliferator-activated receptor gamma(PPARγ)gene on the migration of adipose-derived stem cells(ADSCs)and to explore the possible signaling pathways.Methods Human ADSCs were extracted and cultured in vitro,and different concentrations of PPARγinhibitor bisphenol a diglycidyl ether(BADGE)were applied to ADSCs.The migration of ADSCs was observed by scratch test and Transwell chamber experiment.PCR and western blot were used to detect the expression of MMP2,MMP9,CXCR4 and related proteins.Results The migration ability of ADSCs decreased in both 2 D and 3 D space(P<0.05).The migration of ADSCs in 2 D space:at 72 h,the migration rate of control group was about 63%,and that of 25μm/L group was about 43%,50μm/L group was approximately 24%.Three-dimensional space:the number of ADSCs transfected into the chamber was decreased by 14%and 28%,respectively,compared with the control group by inhibiting the PPARγgene at 24 h in 25μm/L and50μm/L group(P<0.05).The expression of MMP2,MMP9,CXCR4 and related proteins were significantly decreased after inhibition of PPARγgene(P<0.05).Conclusion The migration ability of ADSCs after inhibition of PPARγgene was decreased.The inhibitory effect increased with the increase of inhibitor concentration,and the possible regulatory mechanism was to inhibit the transmission of MMP and CXCR4 signaling pathways.