[Jianwei Shoutai Pill reduces spontaneous abortion in mice by inhibiting Exo-miR-410-3p uptake and promoting trophoblast epithelial-mesenchymal transition, migration, and invasion].

Abstract

Objectives: To investigate the therapeutic effect of Jianwei Shoutai Pill (JSP) on spontaneous abortion (SA) in mice and explore its therapeutic mechanism.

Methods: The miR-410-3p mimic was electroporated into milk exosomes to prepare exosomal miR-410-3p (Exo-miR-410-3p), which was characterized using transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Western blotting. CCK-8, scratch wound healing, and Transwell assays were used to test the effects of control and JSP-medicated rat serum on proliferation, migration, and invasion of trophoblast HTR-8/Svneo cells. Female C57BL/6 mice were divided into SA model group, Model+Exo-miR-410-3p group, Model+Exo-miR-410-3p+P group, and Model+Exo-miR-410-3p+JSP group. The fluorescently labeled exosomes was tracked using in vivo and ex vivo imaging and embryo resorption rate was calculated. RT-qPCR, Western blotting, in situ hybridization, and immunohistochemical studies were carried out for further in vivo verification.

Results: Treatment with Exo-miR-410-3p significantly reduced migration and invasion capacities of HTR-8/Svneo cells, and these inhibitory effects were effectively reversed by JSP intervention. In pregnant C57BL/6 mouse models, JSP treatment prevented Exo-miR-410-3p from reaching the maternal-fetal interface. Consistently, the embryo resorption rate was significantly lower in Exo-miR-410-3p+JSP group than in Exo-miR-410-3p alone group [(20.73±3.77)% vs (74.95±10.09)%, P<0.01]. JSP treatment significant reduced miR-410-3p level and upregulated TRAF6 expression in the female mouse models.

Conclusions: JSP inhibits Exo-miR-410-3p uptake by trophoblast cells, thereby promoting trophoblast EMT, migration, and invasion and reducing embryo resorption in mouse models of spontaneous abortion.