MMP-9在绒毛外滋养细胞诱导内皮细胞凋亡中作用的研究

Effect of MMP-9 on extravillous trophoblasts-induced endothelial apoptosis

目的:探讨绒毛外滋养细胞基质金属蛋白酶-9(MMP-9)的表达对血管内皮细胞凋亡的影响。方法:MMP-9的siRNA转染绒毛外滋养细胞株(TEV-1),利用实时定量逆转录-聚合酶链反应(real time RT-PCR)和ELISA法检测转染前后细胞中MMP-9、FasL基因及蛋白表达的变化;通过trans well共培养技术,研究滋养细胞对内皮细胞凋亡的影响。结果:(1)MMP-9 siRNA转染后24h,滋养细胞MMP-9 mRNA的表达较对照组明显降低(P<0.05),而FasL mRNA表达与对照组无明显差异(P>0.05);转染后48h,细胞培养液中MMP-9蛋白及FasL蛋白表达均较对照组显著降低(P<0.05);(2)与未转染者比较,转染MMP-9 siRNA的滋养细胞与内皮细胞共培养48h后,内皮细胞凋亡率显著降低,差异有显著性(P<0.05)。结论:绒毛外滋养细胞表达MMP-9的水平影响其诱导内皮细胞凋亡的作用,此作用可能与MMP-9调节FasL蛋白的分泌表达有关。

Objective：To investigate the effect of MMP-9 secreted by extravillous trophoblasts on endothelial apoptosis.Methods：MMP-9 siRNA was transfected into extravillous trophoblasts cell line TEV-1.After transfection,the levels of MMP-9,FasL mRNA were assessed by real-time quantitative reverse transcription-polymerase chain reaction（real-time RT-PCR）,and the level of these proteins were measured by ELISA.Then trophoblasts were cocultured with endothelial cells in a transwell system to study endothelial apoptosis.Results：（1）At 24 hour post-transfection,the level of MMP-9 mRNA was significantly decreased as compared with those in the control group（P0.05）.However,FasL mRNA appeared unaffected（P0.05）.48 hours after transfection,the expression level of secreted MMP-9 protein was efficiently decreased,and the secreted FasL protein level decreased correspondingly.（2）At 48hour post coculture,trophoblasts transfected with MMP-9 siRNA led to significant inhibition of endothelial apoptosis as compared with those in the control group（P0.05）.Conclusions：The results suggest that MMP-9 secreted by trophoblasts contributes to endothelial apoptosis via extracellular release of sFasL.