骨间充质干细胞来源的外泌体通过递送 miR-941 抑制 TLR3 缓解米非司酮诱导的人子宫内膜基质细胞损伤。

Exosomes from bone mesenchymal stem cells alleviate mifepristone-induced human endometrial stromal cell injury by inhibiting TLR3 via delivering miR-941.

机构信息

Department of Obstetrics and Gynecology, Shengjing Hospital Affiliated to China Medical University, Shenyang, Liaoning, 110004, China.

出版信息

Physiol Int. 2022 Oct 25;109(4):443-456. doi: 10.1556/2060.2022.00108. Print 2022 Dec 14.

DOI:10.1556/2060.2022.00108

PMID:36422693

Abstract

OBJECTIVE

We aim to investigate the protective effect and underlying mechanisms of BMSCs-exo on human endometrial stromal cells (HESCs) induced by mifepristone in this study.

METHODS

BMSCs-exo were extracted and then identified by transmission electron microscopy and western-blot assay. RT-PCR assay was used to determine the level of miR-941. MiR-941 mimics or inhibitor were transfected into BMSCs and the exosomes were extracted. Then, Cell activity, apoptosis rate, cell migration and invasion, as well as the expression of angiogenic proteins were determined in HESCs stimulated by mifepristone and BMSCs-exo. Next, Dual-luciferase reporting assay was used to verify the targeted binding of miR-941 to TLR3, and the TLR3 expression in HESCs was detected by RT-PCR and western-blot. Finally, TLR3 was overexpressed to evaluate the effects of miR-941 from BMSCs-exo on cell apoptosis, cell invasion and angiogenesis in HESCs induced by mifepristone.

RESULTS

miR-941 was highly expressed in BMSCs-exo. Exosome miR-941 in BMSCs-exo inhibited the cell apoptosis, and promoted cell activity, cell migration, invasion as well as angiogenesis were also improved in HESCs induced by mifepristone. TLR3 was a target of miR-941, which was up-regulated in mifepristonetreated HESCs. We further found that miR-941 derived from BMSCs-exo down-regulated the expression of TLR3 in HESCs treated by mifepristone. In addition, TLR3 overexpression blocked the inhibition of miR-941 on mifepristone-induced cell apoptosis, as well as cell migration and angiogenesis in HESCs.

CONCLUSIONS

Thus, we concluded that BMSCs-exo has protective effect on mifepristone-induced cell damage by delivering miR-941 which targeted TLR3 and regulated cell activity, migration, and angiogenesis in HESCs.

摘要

目的

本研究旨在探讨骨髓间充质干细胞来源外泌体（BMSCs-exo）对米非司酮诱导的人子宫内膜基质细胞（HESCs）的保护作用及其机制。

方法

通过透射电子显微镜和 Western blot 鉴定 BMSCs-exo。采用 RT-PCR 检测 miR-941 的水平。将 miR-941 模拟物或抑制剂转染到 BMSCs 中提取外泌体。然后，检测米非司酮和 BMSCs-exo 刺激的 HESCs 的细胞活力、凋亡率、细胞迁移和侵袭以及血管生成蛋白的表达。接下来，采用双荧光素酶报告基因检测法验证 miR-941 与 TLR3 的靶向结合，采用 RT-PCR 和 Western blot 检测 HESCs 中 TLR3 的表达。最后，过表达 TLR3 以评估 BMSCs-exo 来源的 miR-941 对米非司酮诱导的 HESCs 细胞凋亡、细胞侵袭和血管生成的影响。

结果

miR-941 在 BMSCs-exo 中高表达。BMSCs-exo 中的外泌体 miR-941 抑制了 HESCs 中米非司酮诱导的细胞凋亡，同时也促进了细胞活力、细胞迁移、侵袭和血管生成。TLR3 是 miR-941 的靶基因，在米非司酮处理的 HESCs 中上调。我们进一步发现，BMSCs-exo 来源的 miR-941 下调了米非司酮处理的 HESCs 中 TLR3 的表达。此外，TLR3 过表达阻断了 miR-941 对米非司酮诱导的 HESCs 细胞凋亡、细胞迁移和血管生成的抑制作用。