中国血液净化

中国血液净化 ›› 2019, Vol. 18 ›› Issue (03): 192-196.doi: 10.3969/j.issn.1671-4091.2019.03.013

• 基础研究 • 上一篇    下一篇

MicroRNA-302c通过结缔组织生长因子调控腹膜透析相关腹膜纤维化

李勰家1,周循1,孙林1,刘伏友1,肖力1,刘虹1,张磊1   

  1. 1中南大学湘雅二医院肾内科肾脏疾病与血液净化湖南省重点实验室
  • 收稿日期:2018-10-19 修回日期:2018-12-06 出版日期:2019-03-12 发布日期:2019-03-05
  • 通讯作者: 张磊 zhanglei_xp@163.com E-mail:zhanglei_xp@163.com

MicroRNA- 302C modulates peritoneal dialysis- associated fibrosis by targeting connective tissue growth factor

  • Received:2018-10-19 Revised:2018-12-06 Online:2019-03-12 Published:2019-03-05

PDF

623

摘要： 【摘要】目的通过观察microRNA-302c(miR-302c)对腹膜间皮细胞结缔组织生长因子(connect tissue growth factor,CTGF)表达及上皮细胞-间充质细胞转分化(epithelial-to-mesenchymal transition, EMT)的影响，探讨miR-302c 对腹膜透析(peritoneal dialysis,PD)相关腹膜纤维化的作用及机制。方法收集PD 患者腹膜透析流出液中腹膜间皮细胞，检测miR-302c、CTGF、EMT 及纤维化相关指标的表达，并分析其相关性；体外培养人腹膜间皮细胞株，通过转染慢病毒过表达miR-302c，检测腹膜间皮细胞CTGF、EMT 及纤维化相关指标的变化。结果PD 患者腹膜透析流出液中miR-302c 水平降低(F=443.165，P＜0.001)，与波形蛋白(Vimentin)(r=-0.887，P=0.001)和CTGF(r=-0.840，P=0.002)水平呈负相关，与紧密连接蛋白-1(Zo-1)水平呈正相关(r=0.873，P=0.001)。过表达miR-302c 抑制转化生长因子β1(transforming growth factor-β1,TGF-β1)诱导的人腹膜间皮细胞株E-钙黏蛋白(E-cadherin) (F=13.910，P=0.043)表达下调，α-平滑肌肌动蛋白(α-SMA)(F=11.833，P=0.026)及I 型胶原蛋白(CollagenI) (F=10.673，P=0.031)表达上调，同时也抑制了TGF-β1 诱导的CTGF 表达上调(F=8.340，P=0.044)。结论miR-302c 可能通过CTGF 调控PD 过程中腹膜间皮细胞EMT及纤维化。

关键词: 腹膜透析, microRNA-302c, 上皮细胞-间充质细胞转分化, 腹膜纤维化, 结缔组织生长因子

Abstract: 【Abstract】Object To explore the effect and mechanisms of miRNA-302c on the expression of connective tissue growth factor (CTGF) and epithelial-to-mesenchymal transition (EMT) in peritoneal mesothelial cells from peritoneal dialysis (PD) patients. Methods Human peritoneal mesothelial cells were isolated by collection of overnight peritoneal lavage from PD patients and centrifugation of the lavage. The expression of miR-302c, CTGF and EMT related factors were assayed. Human peritoneal mesothelial cells were transfected with miR-302c and cultured containing 5ng/ml transforming growth factor-β1 (TGF-β1) for 48h, and the expression of CTGF and EMT related factors were then measured by western blot. Results There was a marked reduction of miR-302c level in human peritoneal mesothelial cells from prolonged PD patients (F=443.165, P＜0.001). miR-302c level was negatively correlated with Vimentin (r=-0.887) and CTGF (r=-0.840), and was positively correlated with Zonula occludens-1 (Zo-1) (r=0.873). Over-expression of miR-302c inhibited the down-regulation of E-cadherin (F=13.910，P=0.043) and upregulation of anti-α smooth muscle actin (α-SMA) (F=11.833, P=0.026), collagen I (F=10.673, P=0.031)
and CTGF (F=8.34, P=0.044) induced by TGF-β1 in cultured human peritoneal mesothelial cells. Conclusion MiR-302c is a vital factor protecting peritoneal mesothelial cells from undergoing EMT and fibrosis in PD patients, probably through inhibition of CTGF expression.

Key words: peritoneal dialysis, microRNA-302c, epithelial–to-mesenchymal transition, fibrosis, connective tissue growth factor