首都医科大学学报 ›› 2009, Vol. 30 ›› Issue (2): 161-166.

• 肺循环疾病的基础研究 • 上一篇    下一篇

内皮素通过增强库容性钙内流促进肺血管平滑肌细胞增生

王丛1,2,3, 王辰1,3, 王跃秀1,2,3, 刘杰2,3, 李积凤1,2,3, 王军2,3   

  1. 1. 首都医科大学附属北京朝阳医院呼吸内科,北京呼吸疾病研究所;2. 首都医科大学基础医学院生理教研室;3. 首都医科大学呼吸病学系
  • 收稿日期:2009-01-18 修回日期:1900-01-01 出版日期:2009-04-21 发布日期:2009-04-21
  • 通讯作者: 王辰, 王军

Endothelin-1 Promotes Human Pulmonary Artery Smooth Muscle Cells Proliferation by Up-regulation of Capacitative Ca2+ Entry

WANG Cong1,2,3, WANG Chen1,3, WANG Yue-xiu1,2,3, LIU Jie2,3, LI Ji-feng1,2,3, WANG Jun2,3   

  1. 1. Department of Respiratory Diseases, Beijing Chaoyang Hospital, Capital Medical University, Beijing Institute of Respiratory Medicine;2. Department of Physiology, School of Basic Medical Sciences, Capital Medical University;3. Department of Respirology, Capital Medical University
  • Received:2009-01-18 Revised:1900-01-01 Online:2009-04-21 Published:2009-04-21

摘要:

目的 通过观察内皮素-1(ET-1)诱导人肺动脉平滑肌细胞(HPASMCs)增生的作用机制及细胞内钙离子浓度([Ca2+i)变化,探讨ET-1在肺动脉高压(PAH)发病过程中的作用,为进一步干预治疗提供有效的基础依据。方法 体外培养HPASMCs并分为:正常对照组和内皮素组(ET-1:0.01 μmol/L,0.1 μmol/L,1.0 μmol/L)。分别应用四甲基偶氮唑盐(MTT)法和细胞计数检测细胞的增生情况,荧光钙成像法测定[Ca2+i,运用Real Time-PCR、Western Blotting检测编码钙库操控性钙通道(SOC)的规范瞬时受体电位1(TRPC1)基因及蛋白表达情况。结果 ET-1可促进HPASMCs增生,且ET-1诱导的细胞增生依赖于细胞外Ca2+的内流,ET-1处理的细胞中[Ca2+i和库容性钙内流(CCE)显著升高,同时TRPC1基因转录与蛋白表达也明显增加。结论 ET-1能显著促进肺血管平滑肌细胞增生,其机制可能与上调编码SOC的TRPC1表达、增加库容性钙内流使肺血管平滑肌细胞的[Ca2+i异常升高有关。

关键词: 操纵性通道, 瞬时受体电位通道, 肺动脉平滑肌细胞, 肺动脉高压

Abstract:

Objective This study is designed to investigate the possible signal transduction pathways that related the effect of endothelin-1(ET-1) on HPASMCs proliferation, namely, its effect on the Ca2+ signal pathway. Methods The cultured HPASMCs were divided into following two groups: the control group cultured with smooth muscle basal media (SMBM); the endothelin-1(ET-1) groups incubated with SMBM and ET-1(0.01~1 μmol/L, for 72 h). Cell number was determined with a hemocytometer. MTT assay was used to exam cell viability. [Ca2+i was measured by calcium image. Real Time-PCR was performed to detect the expression of TRPC1 gene. Results The cell viability, cell number, CCE, and TRPC1 mRNA transcription were significantly higher after treatment of ET-1. Conclusion ET-1 induced HPASMCs proliferation via up-regulation of TRP channels expression, CCE and [Ca2+i.

Key words: store-operated channels, transient receptor potential channels, pulmonary artery smooth muscle cells, pulmonary artery hypertension

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