Objective To explore the mechanism of miR-126-5p/Kisspeptin-10(KP-10) participating in endometrial fibrosis through autophagy.

Methods Human endometrial epithelial cells (hEECs) were treated with hypoxia. Transwell method was used to determine the cell migration ability. The expression of miR-126-5p was detected by qRT-PCR. The expressions of autophagy-related proteins (LC3-Ⅰ and LC3-Ⅱ), epithelial-mesenchymal transition (EMT)-related proteins (E-cadherin and N-cadherin) and KP-10 protein were detected by Western blotting. Double luciferase reporter gene analysis was used to confirm the binding relationship between miR-126-5p and KP-10.

Results After hypoxia treatment, the expression of autophagy-related protein LC3-Ⅱ/LC3-Ⅰ gradually increased and reached the peak at 24 h. With the prolongation of hypoxia treatment time, the migration ability of hEECs cells gradually increased (P ＜ 0.05), the expression of E-cadherin decreased (P ＜ 0.05), while the expression of N-cadherin and miR-126-5p increased (P ＜ 0.05). After adding 3-MA or knocking down miR-126-5p, the expression levels of LC3-Ⅱ/LC3-Ⅰ and EMT in hEECs cells induced by hypoxia and cells migration abilities decreased (P ＜ 0.05). Targetscan was used to search the downstream gene of miR-126-5p, and it was found that KP-10 might be one of the targets of miR-126-5p. The results of double luciferase reporter gene analysis showed that miR-126-5p reduced the luciferase activity of cells transfected with KP-10-WT. In addition, after the miR-126-5p was knocked down, the expression of KP-10 protein in hEECs cells was up-regulated (P ＜ 0.05). Compared with the hypoxia + sh-miR-126-5p group, the expression of LC3-Ⅱ/LC3-Ⅰ and EMT in hEECs cells, and cells migration abilities in hypoxia + sh-miR-126-5p + sh-KP-10 group increased (P ＜ 0.05).

Conclusions Knockdown of miR-126-5p reduces hypoxia-induced autophagy and EMT of endometriosis cells by regulating KP-10 signaling, which may become a potential new therapeutic target for endometriosis.