上海交通大学学报(医学版)

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转化生长因子β1对结核性胸膜炎患者间皮细胞上皮-间质转化的影响及机制

王涛,韩娜   

  1. 河北大学附属医院(北院)结核科,保定 071000
  • 出版日期:2016-12-28 发布日期:2016-12-29
  • 通讯作者: 韩娜,电子信箱:1827214480@qq.com。
  • 作者简介:王涛(1980—),男,主治医师,学士;电子信箱:694826581@qq.com。

Effects of transforming growth factor β1 on epithelial-mesenchymal transition in pleural mesothelial cells in patients with tuberculous pleurisy and relevant mechanisms
 

WANG Tao, HAN Na   

  1. Department of Tuberculosis, Affiliated Hospital of Hebei University (North Court), Baoding 071000, China
  • Online:2016-12-28 Published:2016-12-29

摘要:

目的 ·探讨转化生长因子β1(TGF-β1)对结核性胸膜炎患者间皮细胞上皮-间质转化(EMT)的影响及机制。方法 ·选取结核性胸膜炎患者35例,行胸腔镜检查,留取胸腔积液,分离纯化胸膜间皮细胞(PMC),对PMC进行培养并分为4组:空白对照组(仅含细胞培养液)、TGF-β1处理组(加入5 μg/L人重组TGF-β1处理)、PD150606处理组(加入20 mg/L PD150606处理)、联合组(加入5 μg/L人重组TGF-β1和2 mg/L PD150606处理)。培养48 h后,实时荧光定量PCR检测不同处理组细胞calpain-1、CK8、E-cadherin、vimentin和α-SMA基因表达,Western blotting法检测不同处理组细胞中calpain-1、CK8、E-cadherin、vimentin和α-SMA蛋白表达。结果 ·与空白对照组相比,TGF-β1处理组能促使PMC发生EMT,出现间质细胞典型特征,加入calpain 抑制剂PD150606可阻断TGF-β1诱导的PMC上皮-间质转化。与空白对照组相比,TGF-β1处理组calpain-1 基因和蛋白表达量均升高,CK8和E-cadherin基因和蛋白表达量均下降,而vimentin和α-SMA基因和蛋白表达量均上升。加入PD150606后,与TGF-β1处理组相比,联合组calpain-1基因和蛋白表达量均下降,CK8和E-cadherin基因和蛋白表达量均上升,而 vimentin和α-SMA基因和蛋白表达量下降;而与空白对照组相比,联合组calpain-1、CK8、E-cadherin、vimentin和α-SMA基因和蛋白表达量均无差异;PD150606处理组与空白对照组相比均无显著差异。结论 · TGF-β1可能通过上调calpain-1而诱导结核性胸膜炎患者PMC发生EMT。

关键词: 结核性胸膜炎;上皮-间质转化;转化生长因子&beta, 1;钙蛋白酶

Abstract:

Objective · To investigate the effects of transforming growth factor β1 (TGF-β1) on epithelial-mesenchymal transition (EMT) in pleural mesothelial cells in patients with tuberculous pleurisy and relevant mechanisms. Methods · 35 patients with tuberculous pleurisy were enrolled and underwent thoracoscopy test. Pleural effusion specimens were collected and pleural mesothelial cells (PMCs) were Isolated and purified. The PMCs were cultured and divided into different groups, i.e. the blank control group (containing only cell culture medium), the TGF-β1 treatment group (adding 5 μg/L of recombinant human TGF-β1), the PD150606 treatment group (adding 20 mg/L of PD150606), and the combined group (adding 5 μg/L of recombinant human TGF-β1 and 20 mg/L of PD150606). The gene and protein expressions of calpain-1, CK8, E-cadherin, vimentin, and α-SMA were measured using real-time PCR and Western blotting 48h after culture. Results · EMT was induced in PMCs in the TGF-β1 treatment group with typical characteristics of mesenchymal cells compared with the blank control group. Adding calpain inhibitor PD150606 could block the TGF-β1 induced EMT in PMCs. The TGF-β1 treatment group had higher gene and protein expressions of calpain-1, lower gene and protein expressions of CK8 and E-cadherin, and higher gene and protein expressions of vimentin and α-SMA than the blank control group. The combined group had lower gene and protein expressions of calpain-1, higher gene and protein expressions of CK8 and E-cadherin, and lower gene and protein expressions of vimentin and α-SMA than the TGF-β1 treatment group after adding PD150606. The differences in gene and protein expressions of calpain-1, CK8, E-cadherin, vimentin, and α-SMA between the blank control group and the combined group were not significant, as well as between the PD150606 treatment group and the blank control group. Conclusion · TGF-β1 might induce EMT in PMCs in patients with uberculous pleurisy through up-regulation of calpain-1.

Key words: tuberculous pleurisy, epithelial-mesenchymal transition, transforming growth factor β1, calpain