miR-129-5p对尿道下裂术后尿道狭窄瘢痕成纤维细胞增殖的作用机制
Mechanism of miR-129-5p on the proliferation of scar fibroblasts from urethral stricture after hypospadias surgery
胡建军 1赵夭望 1陈毅夫 1刘宇 1何天衢 1王智 1彭柳成1
作者信息
- 1. 中南大学湘雅医学院附属儿童医院(湖南省儿童医院)泌尿外科,长沙 410007
- 折叠
摘要
目的 探讨miR-129-5p调节TGF-β1途径对尿道下裂术后尿道狭窄瘢痕成纤维细胞增殖的作用及机制.方法 选取2021年10月至2021年12月在本院收集的6例尿道下裂患者的术后尿道狭窄瘢痕组织和6例初次手术修剪的正常尿道组织样本,采用qRT-PCR法检测尿道下裂术后尿道狭窄瘢痕组织和正常尿道组织中miR-129-5p和TGF-β1 mRNA的表达;通过TGF-β1诱导尿道成纤维细胞构建过度增殖细胞模型,采用CCK-8法检测miR-129-5p的差异性表达以及对成纤维细胞增殖的影响;采用Western blot检测miR-129-5p的差异性表达对TGF-β1下游蛋白α-SMA和Collagen-Ⅰ表达的影响.结果 尿道下裂术后尿道狭窄瘢痕组织中miR-129-5p和TGF-β1 mRNA的表达水平均明显高于正常尿道组织(均P＜0.01);通过TGF-β1诱导成功构建了尿道成纤维细胞过度增殖细胞模型;在过度增殖细胞模型中,miR-129-5p过表达可对抗TGF-β1诱导成纤维细胞增殖(P＜0.01),miR-129-5p抑制剂能够促进成纤维细胞增殖(P＜0.01),并且使TGF-β1下游蛋白α-SMA和Collagen-Ⅰ的表达上调(均P＜0.01).结论 miR-129-5p通过TGF-β1途径抑制尿道狭窄瘢痕组织的成纤维细胞增殖.
Abstract
Objective To investigate the effect and mechanism of miR-129-5p regulating the expression of TGF-β1 and inhibiting the proliferation of scar fibroblasts from urethral stricture after hy-pospadias surgery.Methods The samples of 6 cases of urethral stricture scar tissue after hypospadias surgery and 6 cases of normal urethral tissue pruned after primary hypospadias surgery were collected in our hosiptal from October 2021 to December 2021.The expression of miR-129-5p and TGF-β1 mRNA in scar tissue and normal urethra tissue after hypospadias surgery was detected by qRT-PCR.The ure-thral fibroblasts were induced by TGF-β1 to construct a hyperproliferative cell model,and the differen-tial expression of miR-129-5p and its effect on the proliferation of fibroblasts were detected by CCK-8.Western blot analysis was performed to detect the effects of miR-129-5p differential expression on the expressions of TGF-β1 downstream proteins α-SMA and Collagenis-Ⅰ.Methods The expression lev-els of miR-129-5p and TGF-β1 mRNA in postoperative urethral stricture scar tissue were significantly higher than those in normal urethral tissue(all P＜0.01).The overproliferation cell model of urethral fibroblasts was successfully constructed by TGF-β1 induction.In the hyperproliferative cell model,the overexpression of miR-129-5p could counter the proliferation of fibroblasts induced by TGF-β1(P＜0.01),and miR-129-5p inhibitors could promote the proliferation of fibroblasts(P＜0.01).Further-more,the expressions of α-SMA and Collagen-I downstream proteins of TGF-β1 were up-regulated(all P＜0.01).Conclusions miR-129-5p can inhibit the growth of fibroblastsin urethral stricture scar tissueby inhibiting TGF-β1.
关键词
尿道下裂/尿道狭窄/miR-129-5p/成纤维细胞/转化生长因子β1Key words
Hypospadias/Urethral Stricture/miR-129-5p/Fibroblasts/Transforming Growth Factor Beta1引用本文复制引用
基金项目
湖南省卫生健康委课题(202204052510)
湖南省儿童泌尿生殖疾病临床医学研究中心(2021SK4017)
出版年
2024
NETL
NSTL
万方数据