Department of Obstetrics and Gynecology, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu, China.
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
J Cell Mol Med. 2021 May;25(10):4583-4595. doi: 10.1111/jcmm.16234. Epub 2021 Apr 9.
Preeclampsia (PE), a common disorder of pregnancy, is characterized by insufficient trophoblast migration and inadequate vascular remodelling, such that promotion of trophoblast proliferation might ameliorate PE. In the current study, we sought to study the underlying mechanism of extracellular vesicle (EV)-derived microRNA-18 (miR-18b) in PE. Human umbilical cord mesenchymal stem cells (HUCMSCs) isolated from placental tissues were verified through osteogenic, adipogenic and chondrogenic differentiation assays. Bioinformatics analyses and dual-luciferase reporter gene assay were adopted to confirm the targeting relationship between miR-18b and Notch2. The functional roles of EV-derived miR-18b and Notch2 in trophoblasts were determined using loss- and gain-of-function experiments, and trophoblast proliferation and migration were assayed using CCK-8 and Transwell tests. In vivo experiments were conducted to determine the effect of EV-derived miR-18b, Notch2 and TIM3/mTORC1 in a rat model of PE, with monitoring of blood pressure and urine proteinuria. TUNEL staining was conducted to observe the cell apoptosis of placental tissues of PE rats. We found down-regulated miR-18b expression, and elevated Notch2, TIM3 and mTORC1 levels in the placental tissues of PE patients compared with normal placenta. miR-18b was delivered to trophoblasts and targeted Notch2 and negatively its expression, whereas Notch2 positively mediated the expression of TIM3/mTORC1. EV-derived miR-18b or Notch2 down-regulation enhanced trophoblast proliferation and migration in vitro and decreased blood pressure and 24 hours urinary protein in PE rats by deactivating the TIM3/mTORC1 axis in vivo. In summary, EV-derived miR-18b promoted trophoblast proliferation and migration via down-regulation of Notch2-dependent TIM3/mTORC1.
子痫前期（PE）是一种常见的妊娠疾病，其特征为滋养细胞迁移不足和血管重塑不足，因此促进滋养细胞增殖可能改善 PE。在本研究中，我们试图研究细胞外囊泡（EV）衍生的 microRNA-18（miR-18b）在 PE 中的潜在机制。从胎盘组织中分离的人脐带间充质干细胞（HUCMSCs）通过成骨、成脂和成软骨分化实验进行验证。采用生物信息学分析和双荧光素酶报告基因实验证实 miR-18b 与 Notch2 之间的靶向关系。通过失活和功能获得实验确定 EV 衍生的 miR-18b 和 Notch2 在滋养细胞中的功能作用，并通过 CCK-8 和 Transwell 试验测定滋养细胞的增殖和迁移。进行体内实验以确定 EV 衍生的 miR-18b、Notch2 和 TIM3/mTORC1 在 PE 大鼠模型中的作用，同时监测血压和尿蛋白尿。TUNEL 染色观察 PE 大鼠胎盘组织的细胞凋亡。我们发现与正常胎盘相比，PE 患者胎盘组织中 miR-18b 表达下调，Notch2、TIM3 和 mTORC1 水平升高。miR-18b 被递送至滋养细胞并靶向 Notch2 并负调控其表达，而 Notch2 则正向调控 TIM3/mTORC1 的表达。EV 衍生的 miR-18b 或 Notch2 下调可通过体内激活 TIM3/mTORC1 轴增强体外滋养细胞的增殖和迁移，并降低 PE 大鼠的血压和 24 小时尿蛋白。总之，EV 衍生的 miR-18b 通过下调 Notch2 依赖性 TIM3/mTORC1 促进滋养细胞的增殖和迁移。
