Hsa-miR-194-5p regulates TRAF6-mediated M1 macrophage apoptosis in recurrent spontaneous abortion

Recurrent spontaneous abortion (RSA) is linked to pro-inflammatory responses driven by macrophage M1 polarization. miR-194-5p can affect the migration and infiltration of macrophages, and significantly inhibit the release of pro-inflammatory cytokines. However, whether miR-194-5p can affect RSA through M1 macrophage-related pathway remains to be further explored. To induce human monocytic leukemia THP-1 into M1 macrophages, PMA and LPS were used. Then detect the effects of transfection with miR-194-5p mimics on the migration, invasion, cell cycle and apoptosis of M1 macrophages. Two databases, DIANA-microT and miRDB, were first used to predict the target gene of miR-194-5p, and TRAF6 was selected as the target gene of miR-194-5p, and then the binding sites of the two were predicted and verified by dual luciferase assay. Transfection of inhibitors, with or without TRAF6 siRNA (si-TRAF6), was performed on M1 macrophages to assess changes in viability, migration, aggressiveness, cell cycle, and apoptosis, as well as TRAF6, NF-κB, and Wnt5a mRNA and protein levels. Compared with the miR-NC group, transfection with the miR-194-5p mimic significantly reduced the viability, migration, and invasion abilities of M1 macrophages, arrested them in the S phase, and promoted apoptosis. miR-194-5p bound to TRAF 3'UTR-WT and reduced the viability, migration ability, and aggressiveness of M1 macrophages, increased apoptosis, and blocked the S phase. miR-194-5p negatively regulated TRAF6, resulting in decreased mRNA and protein levels of NF-κB and Wnt5a. miR-194-5p inhibitors and mimics had opposite effects, but miR-194-5p inhibitor effects could be reversed by si-TRAF6. There is a close association between RSA and M1 macrophage polarization. Furthermore, miR-194-5p inhibits the NF-κB and Wnt5a signaling pathways by negatively regulating TRAF6, thereby impeding the function of M1 macrophages and affecting the occurrence of RSA. These findings provide new therapeutic targets for the prevention, diagnosis, and treatment of RSA.