miR-Let-7b靶向LOX-1减轻子宫内膜纤维化的机制  

Mechanism of miR-Let-7 btargeting LOX-1 to alleviate endometrial fibrosis

作　　者：秦晓洋 戚静宜[1] 王芳[1] 宋瑞芳 QIN Xiaoyang;QIJingyi;WANG Fang;SONG Ruifang(Reproductive Medicine Department of Luoyang Central Hospital,Luoyang 471000,CHN)

机构地区：[1]洛阳市中心医院生殖医学科,河南洛阳471000

出　　处：《河南大学学报（医学版）》2024年第6期407-415,共9页Journal of Henan University:Medical Science

基　　金：河南省医学科技攻关计划(LHGJ20230812)。

摘　　要：目的:检测miR-Let-7b及LOX-1在纤维化子宫内膜中的表达,分析miR-Let-7b靶向LOX-1在减轻子宫内膜纤维化中的作用。方法:分别构建人子宫内膜间质细胞株(hESCs)纤维化模型(TGF-β1-hESCs)及小鼠宫腔粘连模型,qRT-PCR法检测miR-Let-7b及LOX-1表达。慢病毒转染法将TGF-β1-hESCs细胞分为NC组、miR-con组、miR-Let-7bmimic组、miR-Let-7b+pcDNA组和miR-Let-7b+pcDNA-LOX-1组,CCK-8法、EdU法及细胞划痕试验检测各组细胞增殖、迁移能力。将小鼠随机分为假手术组、模型组和治疗组(宫腔注射miR-Let-7bmimic),HE染色观察子宫内膜病理变化,Masson染色法观察子宫内膜纤维化面积,Westernbloting检测各组细胞及子宫内膜组织中α-平滑肌肌动蛋白(α-SMA)、E-钙黏蛋白(E-cadherin)及纤维连接蛋白(fibronectin,FN)表达。双荧光素酶报告基因法验证miR-Let-7b和LOX-1间的靶向关系。结果:TGF-β1-hESCs细胞及宫腔粘连小鼠子宫内膜组织中miR-Let-7b均呈低表达,而LOX-1呈高表达,差异显著(P<0.001)。miR-Let-7bmimic组细胞活力、EdU阳性率、细胞迁移率均低于NC组及miR-con组,差异具有统计学意义(P<0.05)。miR-Let-7b+pcDNA-LOX-1组细胞活力、EdU阳性率、细胞迁移率均高于miR-Let-7b+pcDNA组,细胞内α-SMA、E-cadherin、FN蛋白表达升高,差异有统计学意义(P<0.05)。miR-Let-7b与WT-LOX-1共转染后,miR-Let-7bmimic组细胞荧光素酶活性明显低于miR-con组,差异显著(P<0.001)。与模型组相比,治疗组小鼠子宫内膜腺体数量增多,纤维化面积减少,α-SMA、E-cadherin、FN蛋白表达降低,差异明显(P<0.05)。结论:miR-Let-7b在纤维化子宫内膜中低表达,而LOX-1呈高表达,miR-Let-7b可靶向LOX-1减轻子宫内膜纤维化,机制可能与抑制上皮间充质转化有关。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-1in 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-lbyqRT-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-1group by lentiviral transfection,detect the cell proliferation and migration by CCK-8method,EdU method and cell scratch test.Divide mice into sham surgery group,model group,and treatment group(intrauterine injection of miR-Let-7 bmimic)randomly,observe endometrial pathological changes with HE staining,and endometrial fibrosis area with Masson staining,detect the expressions of a-smooth muscle actin(a-SMA),E-cadherin and fibronectin(FN)in cells and endometrial tissues of each group by Western bloting.Verify the targeting relationship between miR-Let-7 band 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 a-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 a-SMA,E-cadherin and F

关 键 词：miR-Let-7b LOX-1 子宫内膜 纤维化 宫腔粘连 

分 类 号：R715.5[医药卫生—妇产科学]