南方医科大学学报

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (3): 437-446.doi: 10.12122/j.issn.1673-4254.2024.03.04

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二甲双胍通过抑制铁死亡改善PM2.5导致的胎盘滋养细胞功能损伤

李淑贤，于淑平，穆亚铭，王 凯，刘 玉，张美华   

  1. 青岛大学附属山东省妇幼保健院，国家卫生健康委母胎医学重点实验室，山东 济南 250014；山东第二医科大学，山东 潍坊 261053
  • 出版日期:2024-03-20 发布日期:2024-04-03

Metformin ameliorates PM2.5- induced functional impairment of placental trophoblasts by inhibiting ferroptosis

LI Shuxian, YU Shuping, MU Yaming, WANG Kai, LIU Yu, ZHANG Meihua   

  1. Key Laboratory of Maternal & Fetal Medicine of National Health Commission of China，Shandong Provincial Maternal and Child Health Care Hospital Affiliated to Qingdao University, Jinan 250014, China; Shandong Second Medical University, Weifang 261053, China
  • Online:2024-03-20 Published:2024-04-03

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摘要： 目的 探究PM2.5损伤胎盘滋养细胞导致不良妊娠结局的作用机制及二甲双胍的挽救作用。方法 将16只孕鼠随机分为空白组（n=8），PM2.5染毒组（n=8），两组分别在妊娠1.5、7.5、12.5 d通过气管滴注PBS和PM2.5悬浮液。观察PM2.5对孕鼠妊娠结局的影响，并通过HE染色观察小鼠胎盘病理结构，检测胎盘组织铁死亡相关指标。体外构建PM2.5暴露的人类胎盘滋养细胞系HTR8/SVneo细胞染毒模型，利用CCK8实验检测细胞活性；利用EDU染色检测细胞增殖能力；利用划痕实验检测细胞迁移能力；利用Transwell实验检测细胞侵袭能力；利用成管实验检测细胞管生成能力；通过ELISA和Western blotting等方法分析铁死亡相关指标的表达。结果 体内实验显示PM2.5导致胎鼠的质量下降（P<0.001）、数目减少（P<0.01）、死亡率增加（P<0.001），同时胎盘质量也显著减少（P<0.001），HE染色结果显示胎盘的结果受损；组织免疫荧光、ELISA及Western blotting结果表明PM2.5导致胎盘铁死亡的发生。体外实验显示PM2.5通过导致胎盘滋养细胞铁死亡抑制细胞增殖、迁移、侵袭、成管等生物学活性；二甲双胍可显著逆转PM2.5导致的滋养细胞铁死亡，包括细胞内GSH浓度和SOD活性增高（P<0.01）、MDA浓度降低和Fe离子含量降低（P<0.001）、GPX4蛋白和SLC7A11蛋白表达增高（P<0.05）等，同时二甲双胍显著改善PM2.5导致的细胞增值、迁移、侵袭、成管等生物学功能损伤。结论 PM2.5导致孕鼠不良妊娠结局及胎盘滋养细胞铁死亡和功能障碍，应用二甲双胍可有效改善细胞损伤。

关键词: PM2.5；滋养细胞；铁死亡；二甲双胍

Abstract: Objective To investigate the protective effect of metformin against PM2.5-induced functional impairment of placental trophoblasts and explore the underlying mechanism. Methods Sixteen pregnant Kunming mice were randomly assigned into two groups (n=8) for intratracheal instillation of PBS or PM2.5 suspension at 1.5, 7.5, and 12.5 days of gestation. The pregnancy outcome of the mice was observed, and placental zonal structure and vascular density of the labyrinth area were examined with HE staining, followed by detection of ferroptosis-related indexes in the placenta. In cultured human trophoblasts (HTR8/SVneo cells), the effects of PM2.5 exposure and treatment with metformin on cell viability, proliferation, migration, invasion, and tube formation ability were evaluated using CCK8 assay, EDU staining, wound healing assay, Transwell experiment, and tube formation experiment; the cellular expressions of ferroptosis- related proteins were analyzed using ELISA and Western blotting. Results M2.5 exposure of the mice during pregnancy resulted in significantly decreased weight and number of the fetuses and increased fetal mortality with a reduced placental weight (all P<0.001). PM2.5 exposure also caused obvious impairment of the placental structure and trophoblast ferroptosis. In cultured HTR8/SVneo cells, PM2.5 significantly inhibited proliferation, migration, invasion, and angiogenesis of the cells by causing ferroptosis. Metformin treatment obviously attenuated PM2.5-induced inhibition of proliferation, migration, invasion, and angiogenesis of the cells, and effectively reversed PM2.5-induced ferroptosis in the trophoblasts as shown by significantly increased intracellular GSH level and SOD activity, reduced MDA and Fe2+ levels, and upregulated GPX4 and SLC7A11 protein expression (P<0.05 or 0.01). Conclusion PM2.5 exposure during pregnancy causes adverse pregnancy outcomes and ferroptosis and functional impairment of placental trophoblasts in mice, and metformin can effectively alleviate PM2.5-induced trophoblast impairment.

Key words: PM2.5; trophoblasts; ferroptosis; metformin