秦喆，侯海燕，徐忠伟，张立文，韩斌，陈亚琼，吴思雨，姚婷

·论著·

PM2.5对人绒毛膜外滋养层细胞HTR8-SVneo迁移与侵袭力的影响及机制研究

秦喆，侯海燕，徐忠伟，张立文，韩斌，陈亚琼，吴思雨，姚婷

目的：探讨PM2.5对人绒毛膜外滋养细胞HTR8-SVneo存活率以及迁移与侵袭能力的影响，寻找PM2.5致不良妊娠结局可能的机制。方法：不同浓度PM2.5染毒HTR8-SVneo细胞建立实验模型，CCK8（Cell Counting Kit-8）法分析细胞增殖情况，最终选取0、30、60、120和200μg/mL 5个浓度进行后续实验，其中以0 μg/mL染毒组为对照组。激光全息细胞分析及成像系统分析细胞迁移力；Transwell侵袭实验分析细胞侵袭力；qPCR和Western Blot分别检测基质金属蛋白酶2（MMP2）、MMP9及其组织抑制剂TIMP1、TIMP2的mRNA和蛋白表达水平；明胶酶谱法检测细胞分泌的明胶酶MMP2和MMP9活性。结果：PM2.5对HTR8-SVneo细胞增殖有显著抑制作用（P＜0.05）；PM 2.5使HTR8-SVneo细胞迁移距离和穿膜细胞数均减少（P＜0.05）；qPCR和Western Blot结果显示，PM2.5使HTR8-SVneo细胞MMP2、MMP9、TIMP1及TIMP2的mRNA及蛋白表达水平升高（P＜0.05），且MMPs升高程度低于TIMPs；明胶酶谱结果显示，HTR8-SVneo细胞MMP9活性在30、60、120和200μg/m L染毒组均升高（P＜0.05）。结论：PM2.5在造成HTR8-SVneo细胞损伤的同时，可能通过打破MMPs/TIMPs的动态平衡影响人绒毛膜外滋养细胞HTR8-SVneo的迁移与侵袭力，从而造成不良妊娠结局，而具体信号机制有待深入研究。

空气污染物；滋养层；细胞迁移分析；基质金属蛋白酶2；基质金属蛋白酶9；金属蛋白酶1组织抑制剂；金属蛋白酶2组织抑制剂

【Abstract】Objective:To investigate the effect of PM2.5 on the survival rate，themigration ability and the invasion ability of human chorionic trophoblast cell line，HTR8-SVneo，and to explore the possiblemechanism of the adverse pregnancy outcome induced by PM2.5.Methods:The in vitro cultured HTR8-SVneo cells were treated with PM2.5.The cell proliferation was analysed by CCK8.Five concentrations of 0，30，60，120 and 200 μg/mL were selected.The cell migration was detected by Laser holographic cell analysis and imaging system，while the cell invasion by the Transwell experiment.ThemRNA and protein expressions ofMMP2，MMP9，TIMP1 and TIMP2 were measured by qPCR and Western Blot，while the activities of MMP2 and MMP9 by a gelatin zymography.Resu lts:The proliferation of HTR8-SVneo cellswas significantly inhibited by PM 2.5 treatment（P＜0.05），while themigration distance and the number of cells penetrating themembrane were decreased（P＜0.05）. The mRNA and protein expressions of MMP2，MMP9，TIMP1 and TIMP2 were significantly increased in PM2.5 exposed groups（P＜0.05），and the increased degree ofMMPswas lower than TIMPs.The activities ofMMP9 in all PM2.5 treatment groups were significantly increased when compared with the control group（P＜0.05）. Conclusions:PM 2.5 can induce the cellular damage of HTR8-SVneo cells，while PM2.5 can decrease the cell migration and invasion by inducing the imbalance between MMPs and TIMPs.These pathophysiological changes are related with the adverse pregnancy outcomes.

【Keywords】Air pollutants；Trophoblasts；Cell migration assays；Matrix metalloproteinase 2；Matrix metalloproteinase9；Tissue inhibitorofmetalloproteinase-1；Tissue inhibitorofmetalloproteinase-2

（J Int Reprod Health/Fam Plan，2016，35：357-361，386）

目前，直径≤2.5μm的可吸入颗粒物（PM2.5）对人类健康的影响成为了研究热点，除造成呼吸系统、心血管系统疾病外，大量流行病学研究表明PM2.5与不良妊娠结局有关［1］。妊娠成功是依赖于滋养细胞适度侵入蜕膜组织促进胚胎着床、重建细胞外基质及血管以维持胎盘正常生理功能。……