Mechanism of miR-Let-7b targeting LOX-1 to alleviate endometrial fibrosis
Objective:To detect the expression of miR-Let-7b and LOX-1 in fibrotic endometrium,and analyze the role of miR-Let-7b targeting LOX-1 in reducing endometrial fibrosis.Methods:Construct human endometrial stromal cell line(hESCs)fibrosis model(TGF-β1-hESCs)and mouse uterine adhesion model,respectively,and detecte the expression of miR-Let-7b and LOX-1 by qRT-PCR.Divide TGF-β1-hESCs cells into NC group,miR-con group,miR-Let-7b mimic group,miR-Let-7b+pcDNA group and miR-Let-7b+PCDNA-LOX-1 group by lentiviral transfection,detect the cell proliferation and migration by CCK-8 method,EdU method and cell scratch test.Divide mice into sham surgery group,model group,and treatment group(intrauterine injection of miR-Let-7b mimic)randomly,observe endometrial pathological changes with HE staining,and endometrial fibrosis area with Masson staining,detect the expressions of α-smooth muscle actin(α-SMA),E-cadherin and fibronectin(FN)in cells and endometrial tissues of each group by Western bloting.Verify the targeting relationship between miR-Let-7b and LOX-1 using the dual luciferase reporter gene method.Results:The expression of miR-Let-7b was low in TGF-β1-hESCs cells and endometrial tissue of uterine adhesion mice,while the expression of LOX-1 was high,with significant differences(P＜0.001).The cell viability,EdU positive rate and cell mobility of miR-Let-7b mimic group were lower than those of NC group and miR-con group,and the differences were statistically significant(P＜0.05).The cell viability,EdU positive rate and cell mobility of miR-Let-7b+PCDNA-Lox-1 group were higher than those of miR-Let-7b+pcDNA group,and the expressions of α-SMA,E-cadherin and FN protein were increased,with statistical significance(P＜0.05).After co-transfection of miR-Let-7b and WT-LOX-1,the cell luciferase activity of miR-Let-7b mimic group was significantly lower than that of miR-con group(P＜0.001).The treatment group mice showed an increase in the number of endometrial glands and a decrease in fibrotic area,the expression of α-SMA,E-cadherin and FN protein when compared with the model group,the difference was significant(P＜0.05).Conclusion:miR-Let-7b is low expressed in fibrotic endometrium,while LOX-1 is highly expressed.miR-Let-7b can target LOX-1 to alleviate endometrial fibrosis,and the mechanism may be related to inhibiting epithelial mesenchymal transition.
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