Conclusion
MSC-exo ameliorated TGF-β1-induced endometrial fibrosis. RNA sequencing, bioinformatic analysis, and functional experiments revealed that miR-145-5p may exert its action through the P62-dependent autophagy pathway.
Methods
MSC-exo were isolated and identified based on particle size and protein marker detection. Cell counting kit-8, flow cytometry, and western blotting were used to determine the effects of MSC-exo on cell function and fibrosis in human endometrial epithelial cells (hEECs). Subsequently, we sequenced and annotated the small RNA in MSC-exo and TGF-β1-induced MSC-exo to screen for differentially expressed (DE) miRNAs. After the prediction and functional enrichment of target genes of DE miRNAs, key genes were selected for functional experiments.
Objective
Mesenchymal stem cell (MSC)-derived exosomes (MSC-exo) can treat reproductive disorders. However, the action of microRNAs (miRNAs) in this mechanism has yet to be systematically investigated. This study aimed to explore the effect of MSC-exo on TGF-β1-induced endometrial fibrosis in intrauterine adhesions and elucidate the regulatory mechanisms involved in key genes by comparing miRNA expression profiles.
Results
TGF-β1 inhibited the proliferation of hEECs and promoted apoptosis and fibrosis. However, these effects were significantly reversed by the addition of MSC and MSC-exo. Fifteen DE miRNAs were identified by comparing the miRNA profiles of MSC-exo and TGF-β1-induced MSC-exo. Among these, miR-145-5p was found to be significantly upregulated in TGF-β1-induced MSC-exo. Furthermore, the addition of miR-145-5p mimic was found to reverse fibrosis in hEECs while promoting the expression of key autophagy protein P62.