子痫前期患者血清对滋养-平滑肌细胞共培养体系平滑肌细胞凋亡的影响

Effect of Preeclampsia Serum on Human Umbilical Artery Smooth Muscle Cell Apoptosis in a Model of Cytotrophoblast Coculture with Human Umbilical Artery Smooth Muscle Cell

目的:探讨子痫前期血清对滋养-平滑肌细胞共培养体系滋养细胞侵袭力改变及对平滑肌细胞凋亡的影响。方法:采用组织块贴壁法培养人孕早期细胞滋养细胞(CTB)和人脐动脉平滑肌细胞(HUASMC),传代后建立滋养细胞与HUASMC共培养模型并分别加入正常和子痫前期患者静脉血清培养24小时并设立相对应的单培养组;用不同的方法检测细胞滋养细胞的侵袭能力、HUASMC活力、HUASMC凋亡率、滋养细胞基质金属蛋白酶MMP-2mRNA、MMP-9mRNA、人脐静脉内皮细胞自杀相关因子(Fas)及HUASMC的Fas配体(Fasl)mRNA的表达;HUASMC的Fas蛋白的表达。结果:子痫前期患者血清能显著抑制共培养体系中的细胞滋养细胞的侵袭能力,上调HUASMC活力,降低HUASMC早期凋亡率,降低细胞滋养细胞MMP-2mRNA、MMP-9mRNA、FasmRNA及HUASMC的FaslmRNA和Fas蛋白水平的表达。结论:子痫前期血清可能通过降调细胞滋养细胞MMP-2、MMP-9表达而影响滋养-平滑肌细胞共培养体系滋养细胞的侵袭能力,子痫前期血清培养滋养-平滑肌细胞共培养体的Fas/Fasl的表达异常可能与HUASMC凋亡下降有关。提示滋养细胞MMP-2、MMP-9、Fas/Fasl系统的表达异常可能参与了子痫前期子宫螺旋动脉重铸障碍的病理过程。

Objective：To investigate cytotrophoblast invasive ability and human umbilical artery smooth muscle cell apoptosis in the model of cytotrophoblast coculture with human umbilical artery smooth muscle cell using preeclampsia serum. Methods：Cytotrophoblast（CTB） and human umbilical artery smooth muscle cell （HUASMC） were collected by explant culture. The model of CTB coculture with HUASMC was constructed and incubated with normal serum or preeclampsia serum for 24 hours,as controls, CTB or HUASMC were monocultured and treated in other ways the same as the coculture situation. Analysis of the invasive ability of CTB,the activity and apoptosis of HUASMC, and the expressions of MMP-2, MMP-9, Fas, FaslmRNA and Fas protein. Results：In the model of CTB coculture with HUASMC,the preeclampsia serum can effectively inhibit the invasive ability of cytotrophoblast,increase the activity of HUASMC,decrease apoptotic rate of HUASMC, and decrease the expressions of MMP-2, MMP-9, FasmRNA in CTB, also decrease the expressions of Faslm- RNA and Fas protein in HUASMC. Conclusions ：The preeclampsia serum can affect the invasive ability of cytotrophoblast in a model of CTB coculture with HUASMC via down regulate the expression of MMP-2,MMP-9. The decrease of apoptosis of HUASMC may relate with the abnormal expression of Fas/Fasl. This data provide a rationale that the abnormal expression of MMP-2, MMP-9 and Fas/Fasl system in CTB may participate in the remodelina defects of the uterine spiral arteries in DreeclamDsia.