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miR-410-5p/ITGA6 轴通过调节滋养细胞的生物学功能参与复发性流产的发病机制。

The miR-410-5p /ITGA6 axis participates in the pathogenesis of recurrent abortion by regulating the biological function of trophoblast.

机构信息

Reproductive Medical Center, Renmin Hospital of Wuhan University and Hubei Clinic Research Center for Assisted Reproductive Technology and Embryonic Development, Wuhan, Hubei, China.

Department of Spine Surgery & Musculoskeletal Tumor, Zhongnan Hospital of Wuhan University, Wuhan, Hubei, China.

出版信息

J Reprod Immunol. 2022 Aug;152:103647. doi: 10.1016/j.jri.2022.103647. Epub 2022 May 23.

Abstract

The purpose of this study was to determine the regulation of the miR-410-5p /ITGA6 axis on the biological functions of trophoblast cells and the mechanism involved in recurrent spontaneous abortion(RSA). We used qRT-PCR and Western blotting to quantify the expression levels of Mir-410-5p and ITGA6 in placenta of RSA and normal, and found that compared with normal placenta, the placenta of RSA patients showed higher miR-410-5p and lower ITGA6 expression. Dual luciferase reporter gene assay confirmed the binding of miR-410-5p to ITGA6. The expression of miR-410-5p and ITGA6, and proliferation, apoptosis, invasion and migration of trophoblast cells and the effect on the polarization of M2 macrophages were detected in the trophoblast derived cell lines HTR8/Svneo transfected with miR-410-5p mimic, sh-miR-410-5p and si-ITGA6 respectively. Meanwhile, the molecular mechanism of ITGA6 regulation on trophoblast cells was explored. Transfection with miR-410-5p mimic or si-ITGA6 attenuated the proliferation, migration and invasion and induced apoptosis of HTR-8/SVneo cells. Transfection of sh-miR-410-5p promoted proliferation, migration and invasion, and weakened apoptosis of HTR-8/SVneo cells. In addition, overexpression of miR-410-5p in trophoblast cells inhibited the polarization of M2 macrophages, while knockdown of miR-410-5p was beneficial to recruitment of trophoblast cell and promoted the polarization of M2 macrophages. ITGA6 may affect the biological functions of trophoblast cells by regulating PI3K/AKT and MAPK signaling pathways. In conclusion, miR-410-5p mediates trophoblast cell proliferation, apoptosis, invasion and migration through regulating ITGA6 expression.

摘要

本研究旨在确定 miR-410-5p/ITGA6 轴对滋养层细胞生物学功能的调节作用及其在复发性自然流产(RSA)中的作用机制。我们使用 qRT-PCR 和 Western blot 技术定量检测了 RSA 和正常胎盘组织中 Mir-410-5p 和 ITGA6 的表达水平,发现与正常胎盘相比,RSA 患者胎盘组织中 miR-410-5p 表达升高,ITGA6 表达降低。双荧光素酶报告基因实验证实了 miR-410-5p 与 ITGA6 的结合。分别转染 miR-410-5p 模拟物、sh-miR-410-5p 和 si-ITGA6 后,检测滋养层来源细胞系 HTR8/Svneo 中 miR-410-5p 和 ITGA6 的表达以及滋养层细胞的增殖、凋亡、侵袭和迁移,并观察对 M2 巨噬细胞极化的影响。同时,探讨了 ITGA6 对滋养层细胞的调控作用的分子机制。转染 miR-410-5p 模拟物或 si-ITGA6 可减弱 HTR-8/SVneo 细胞的增殖、迁移和侵袭,诱导细胞凋亡。转染 sh-miR-410-5p 则促进 HTR-8/SVneo 细胞的增殖、迁移和侵袭,减弱细胞凋亡。此外,在滋养层细胞中过表达 miR-410-5p 可抑制 M2 巨噬细胞的极化,而敲低 miR-410-5p 则有利于滋养层细胞的募集,并促进 M2 巨噬细胞的极化。ITGA6 可能通过调节 PI3K/AKT 和 MAPK 信号通路影响滋养层细胞的生物学功能。综上所述,miR-410-5p 通过调节 ITGA6 的表达来介导滋养层细胞的增殖、凋亡、侵袭和迁移。

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