Electroacupuncture ameliorates embryo implantation dysfunction in mice via miR-30c-5p.

OBJECTIVE

To investigate the intrinsic mechanism by which electroacupuncture (EA) treats embryo implantation dysfunction in mice from the perspective of exosomal miRNAs.

METHODS

Human trophoblast cells (HTR-8/Svneo) were co-cultured in vitro with endometrial cells (Ishikawa and HEC-1-A), and exosomes derived from endometrial cells were isolated to treat HTR-8/Svneo cells. Cell proliferation and invasion were assessed by EdU staining and Transwell assays. miRNA sequencing was performed on exosomes from the two endometrial cell lines to identify differentially expressed miRNAs. A mifepristone-induced mouse model of implantation dysfunction was established and treated with EA. The number of implantation sites was counted, pathological changes in the endometrium were observed by hematoxylin-eosin (HE) staining, and miR-30c-5p expression was detected by qPCR. miR-30c-5p was inhibited in HTR-8/Svneo cells, and changes in cell proliferation, invasion, and phosphorylation of FAK and JNK were evaluated.

RESULTS

Communication existed between HTR-8/Svneo cells and endometrial cells, and co-culture with Ishikawa cells significantly promoted the proliferation and invasion of HTR-8/Svneo cells. Exosomes derived from both Ishikawa and HEC-1-A cells promoted HTR-8/Svneo cell proliferation and invasion. High-throughput sequencing showed that miR-30c-5p was expressed at a low level in Ishikawa cell-derived exosomes but at a high level in HEC-1-A cell-derived exosomes. In vivo experiments demonstrated that EA reduced miR-30c-5p expression in the endometrium of mice with implantation dysfunction, showed a trend of increasing implantation sites, and alleviated endometrial injury. Inhibition of miR-30c-5p in HTR-8/Svneo cells promoted cell proliferation and invasion and increased the phosphorylation of FAK and JNK.

CONCLUSION

EA may ameliorate embryo implantation dysfunction in mice by downregulating miR-30c-5p in the endometrium.