miR-34a-5p靶向Smad4抑制子宫内膜纤维化的机制研究  

Inhibitory effect of miR-34a-5p on fibrosis of endometrial by down-regulating Smad4

作　　者：范李静 张苗 郑瑛红 刘文杰 范小斌 Fan Lijing;Zhang Miao;Zheng Yinghong(Department of Obstetrics and Gynecology,the Affiliated Hospital of Northwest University,Xi'an NO.3 Hospital,Xi'an 710021)

机构地区：[1]西北大学附属医院西安市第三医院妇产科,西安710021

出　　处：《现代妇产科进展》2022年第9期678-682,共5页Progress in Obstetrics and Gynecology

基　　金：陕西省重点研发计划(No:2020SF-054)。

摘　　要：目的:探讨miR-34a-5p在子宫内膜纤维化中的作用及机制。方法:收集正常及宫腔粘连患者的子宫内膜组织,实时定量PCR检测子宫内膜组织中miR-34a-5p表达水平。子宫内膜间质细胞(hESC)转染miR-34a-5p mimics(或mimics对照)或Smad4过表达质粒,并用转化生长因子-β1(TGF-β1)处理。CCK-8检测细胞增殖能力。实时定量PCR和Western blot法检测纤维化标志物α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)和胶原蛋白Ⅰ(COL1A1)的mRNA和蛋白表达水平。生物信息学软件预测miR-34a-5p的靶基因并用双荧光素酶报告基因方法验证。结果:与正常患者相比,宫腔粘连患者子宫内膜组织中miR-34a-5p水平降低。TGF-β1处理后可显著降低hESC细胞中miR-34a-5p水平,增加细胞增殖率。与阴性对照NC相比,转染miR-34a-5p mimics可显著降低hESC细胞的增殖率,以及α-SMA、FN和COL1A1的mRNA和蛋白表达水平。Smad4经鉴定是miR-34a-5p的靶基因。过表达Smad4可逆转miR-34a-5p mimics抗增殖及抗纤维化作用,表现为hESC细胞增殖率增加,α-SMA和COL1A1表达水平增加。结论:miR-34a-5p通过下调Smad4抑制TGF-β1诱导的hESC细胞增殖和纤维化。Objective:To investigate the role of miR-34 a-5 p in endometrial fibrosis and the underling mechanism.Methods:Endometrial tissues from normal and patients with intrauterine adhesions were collected,and the expression of miR-34 a-5 p in endometrial tissues was detected by quantitative real-time.hESC cells were transfected with miR-34 a-5 p mimics/a mimics control or Smad4 overexpression plasmid and were treated with transforming growth factorβ1(TGF-β1).Cell proliferation ability was detected by CCK-8.The mRNA and protein expression levels of fibrosis markersα-smooth muscle actin(α-SMA),fibronectin(FN)and collagen I(collagen I)were detected by quantitative real-time PCR and Western blot.The target genes of miR-34 a-5 p were predicted and validated with the dual-luciferase reporter gene method.Results:Compared with normal patients,the level of miR-34 a-5 p was decreased in endometrial tissue of patients with intrauterine adhesions.TGF-β1 treatment significantly downregulated the level of miR-34 a-5 p in hESC cells and induced the cell proliferation rate.Compared with the mimics control,transfection of miR-34 a-5 p mimics significantly reduced the proliferation rate of hESC cells,the mRNA and protein expression levels ofα-SMA,FN and COL1 A1.Smad4 was identified as a target gene of miR-34 a-5 p and overexpression of Smad4 significantly reversed the anti-proliferative and anti-fibrotic effectsof miR-34 a-5 p mimics,indicated by increased proliferation rate of hESC cells and increased expression levels ofα-SMA and COL1 A1.Conclusion:miR-34 a-5 p inhibits TGF-β1-induced proliferation and fibrosis in hESC cells by downregulating Smad4.

关 键 词：miR-34a-5p 宫腔粘连 纤维化 SMAD4 

分 类 号：R711.4[医药卫生—妇产科学]