滋养层细胞衍生的细胞外囊泡通过调节巨噬细胞极化促进子痫前期。

Trophoblast-Derived Extracellular Vesicles Promote Preeclampsia by Regulating Macrophage Polarization.

机构信息

Assisted Reproduction Unit, Department of Obstetrics and Gynecology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China (X.L., H.F., C.Y., J.W., A.Y., Z.S., X.J., L.J., S.Z.).

Department of Obstetrics and Gynecology, Key Laboratory of Reproductive Dysfunction, Management of Zhejiang Province, China (X.L., H.F., C.Y., J.W., A.Y., Z.S., X.J., L.J., S.Z.).

出版信息

Hypertension. 2022 Oct;79(10):2274-2287. doi: 10.1161/HYPERTENSIONAHA.122.19244. Epub 2022 Aug 22.

DOI:10.1161/HYPERTENSIONAHA.122.19244

PMID:35993233

Abstract

BACKGROUND

Systemic inflammation caused by dysfunctional macrophages is a crucial pathogenetic event in preeclampsia (PE). Trophoblast-derived extracellular vesicles (T-EVs) are potent immune cell signaling modulators in pregnancy. Herein, we aimed to investigate T-EVs' effect and mechanism on macrophage polarization and its role in PE pathogenesis, which remain unclear.

METHODS

Flow cytometry and immunochemistry were used to determine placental macrophage phenotypes. T-EVs were immuno-isolated via placental alkaline phosphatase antibody and identified by transmission electron microscopy and nanoparticle tracking analysis. Quantitative real-time polymerase chain reaction and flow cytometry were used to examine the effects of T-EVs on macrophage polarization, and correlation analysis of T-EVs lipidomics and macrophages transcriptome were performed to explore how T-EVs modulate macrophages. Animal experiments were established to investigate the relationship among PE, T-EVs, and macrophages.

RESULTS

Macrophages shift from the M2 to M1 phenotype in the preeclamptic placenta. Also, T-EVs from women with PE (PE-EVs) significantly upregulated M1 gene markers and significantly downregulated CD163 expression in macrophages compared with T-EVs in women with normal pregnancies (NP-EVs). Mechanistically, correlation analysis with T-EVs lipidome and the transcriptome of macrophages treated with PE-EVs or NP-EVs indicated that 37 lipids altered in PE-EVs considerably affected classical inflammatory biological pathways in macrophages. Finally, animal experiments revealed that PE-EVs triggered PE-like symptoms in pregnant mice, which were alleviated after macrophage depletion.

CONCLUSIONS

T-EVs from women with PE could promote preeclampsia by inducing macrophage imbalance polarization, signifying a potential novel interventional target for the prevention and management of PE.

摘要

背景

功能失调的巨噬细胞引起的全身炎症是子痫前期（PE）发病机制中的关键事件。滋养层衍生的细胞外囊泡（T-EVs）是妊娠中强有力的免疫细胞信号调节剂。在此，我们旨在研究 T-EVs 对巨噬细胞极化的影响和作用机制，以及其在 PE 发病机制中的作用，这些方面仍不清楚。

方法

采用流式细胞术和免疫组化方法确定胎盘巨噬细胞表型。通过胎盘碱性磷酸酶抗体免疫分离 T-EVs，并通过透射电子显微镜和纳米颗粒跟踪分析进行鉴定。采用定量实时聚合酶链反应和流式细胞术研究 T-EVs 对巨噬细胞极化的影响，并进行 T-EVs 脂质组学与巨噬细胞转录组的相关性分析，以探讨 T-EVs 如何调节巨噬细胞。建立动物实验以研究 PE、T-EVs 和巨噬细胞之间的关系。

结果

PE 胎盘的巨噬细胞从 M2 表型向 M1 表型转变。与正常妊娠妇女（NP-EVs）的 T-EVs 相比，PE 妇女（PE-EVs）的 T-EVs 显著上调了巨噬细胞中 M1 基因标志物的表达，并显著下调了 CD163 的表达。从机制上讲，与 T-EVs 脂质组和用 PE-EVs 或 NP-EVs 处理的巨噬细胞转录组的相关性分析表明，PE-EVs 中改变的 37 种脂质显著影响了巨噬细胞中经典炎症生物学途径。最后，动物实验表明，PE-EVs 可在妊娠小鼠中引发类似 PE 的症状，而在耗尽巨噬细胞后可缓解这些症状。