【摘要】 目的:探讨早孕蜕膜基质细胞培养液(DSCM)对滋养细胞增殖、凋亡以及浸润能力的影响及对浸润功能的作用机制。方法:原代分离、培养并鉴定人早孕蜕膜基质细胞,收集并制备成不同浓度的DSCM,采用噻唑兰(MTT)、Annexin V-FITC、Transwell方法检测不同浓度的DSCM对滋养细胞增殖、凋亡、浸润能力的影响,采用RT-PCR及明胶酶谱法检测滋养细胞基质金属蛋白酶(MMPs)的改变。结果:得到纯度大于90%的蜕膜基质细胞。随着DSCM的浓度增高(5%DSCM,20%DSCM,50%DSCM,100%DSCM)细胞的吸光度值增高、凋亡率减少、浸润细胞数增多(P<0.05)。不同浓度的DSCM组MMP-2及MMP-9 mRNA、蛋白的变化与对照组相比差异均有统计学意义(P<0.05)。结论:早孕DSCM可以促进滋养细胞增殖,抑制凋亡,并通过上调滋养细胞对MMP-2与MMP-9的表达来促进其浸润功能。更多还原

【Abstract】 Objective: To investigate the impact of conditioned medium of decidual stromal cell culture( DSCM) on proliferation,apoptosis and invasion of trophoblast cells. Methods: Isolution,culturing and indentifying the purity of decidual stromal cells from healthy women of early pregnancy to prepare different concentrations of DSCM( 5%,20%,50% and 100%); Proliferation was evaluated by MTT assay; Cell apoptosis was examined by flow cytometry using annexin V-FITC/PI staining; Cell invasion was assayed using a Transwell chamber; RT-PCR and gelatin zymography were used to explore the mechanism of cell invasion.Results: Using this method we obtained a purity over 90% of decidual stromal cells.With the concentration of DSCM increased( 5%,20%,50%,100%),the OD value of the MTT assay has elevated and the rate of apoptosis declined,the infiltrating cell increased as well( P <0. 05); Compared with control group,DSCM treatment led to a significant increase in the m RNA and protein expression of MMP-2/9( P<0. 05).Conclusions: Decidual microenviroment may promote trophoblast proliferation and inhibite its apoptosis besides could enhance its ability of invasion by altering the expression of MMP-2 and MMP-9.更多还原