首都医科大学学报 ›› 2013, Vol. 34 ›› Issue (2): 224-231.doi: 10.3969/j.issn.1006-7795.2013.02.013

• 基础研究 • 上一篇    下一篇

西地那非抑制低氧对人肺动脉平滑肌细胞电压依赖性钾通道的下调作用

毛婷1,2, 王跃秀1,2, 刘杰1,2, 李晶1,2, 王军1,2   

  1. 1. 首都医科大学基础医学院生理学教研室, 北京 100069;
    2. 首都医科大学附属北京朝阳医院 北京市呼吸和 肺循环疾病重点实验室, 北京 100020
  • 收稿日期:2012-10-24 出版日期:2013-04-21 发布日期:2013-04-17
  • 通讯作者: 毛婷, 王军 E-mail:wang_jun@ccmu.edu.cn; yuexiuw@126.com
  • 基金资助:

    国家973计划项目(2009CB522107),国家自然科学基金(81070042),重大国际合作研究基金项目(30810103904),海外及港澳学者合作研究基金(81228001),北京呼吸与肺循环疾病重点实验室开放课题(RPCD201201,2011HXFB04),首都医科大学基础临床合作课题(12JL15),北京市教委科技发展计划项目(KM201310025004),宁夏自然科学基金(NZ1220)。

Sildenafil inhibits hypoxia-induced voltage-dependent potassium channels downregulation in human pulmonary artery smooth muscle cells

MAO Ting1,2, WANG Yuexiu1,2, LIU Jie1,2, LI Jing1,2, WANG Jun1,2   

  1. 1. Department of Physiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China;
    2. Beijing Key Laboratory of Respiratory and Pulmonary Circulation, Beijing Chaoyang Hospital, Capital Medical University, Beijing 100020, China
  • Received:2012-10-24 Online:2013-04-21 Published:2013-04-17
  • Supported by:

    This study was supported by the 973 program(2009CB522107), National Natural Science Foundation of China(81070042), Major International Collaborative Research Foundation of China(30810103904), Overseas and Hong Kong and Macao Scholars of Collaborative Research Foundation of China(81228001), Beijing Key Laboratory of Respiratory and Pulmonary Circulation Disorders in Capital Medical University(RPCD201201, 2011HXFB04), Basis-clinical Research Co-operation Fund of Capital Medical University(12JL15), The Fundation of Scientific Research Common Program of Beijing Municipal Commission of Education(KM201310025004), Natural Science Foundation of Lingxia(NZ1220).

摘要:

目的 探讨西地那非对低氧下调人肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)电压依赖性钾通道(voltage-dependent potassium channels,Kv)的干预作用。方法 应用膜片钳技术记录PASMCs K+电流;观察低氧刺激前后钾离子(K+)电流的变化以及西地那非(sildenafil)的干预作用;通过特异性Kv1.5抗体透析,分析西地那非作用于人PASMCs Kv分子靶点即通道亚单位;运用Real-time PCR及Western blotting技术检测低氧刺激前后及西地那非对人PASMCs上的Kv1.5通道基因转录水平和蛋白表达水平的影响。结果 低氧明显抑制了人PASMCs细胞膜上的全细胞K+电流;而西地那非则可以对抗低氧对全细胞K+电流的抑制作用;而且西地那非对低氧下人PASMCs Kv的调控作用靶点主要是Kv1.5;同时西地那非部分恢复了低氧抑制的细胞上的Kv1.5 mRNA和蛋白表达。结论 西地那非能够抑制低氧对人肺动脉平滑肌细胞电压依赖性钾通道功能与表达的下调作用。

关键词: 西地那非, 电压依赖性钾通道, 人肺动脉平滑肌细胞

Abstract:

Objective To investigate the effect of sildenafil on hypoxia-induced voltage-dependent potassium channels(Kv) downregulation in human pulmonary artery smooth muscle cells(PASMCs). Methods Patch clamp techniques were applied to record K+ currents from human PASMCs cultured with SMBM(2% FBS) in normoxia or hypoxia(3% O2) in the absence or presence of sildenafil(100 nM) for 72 h. Furthermore, the effect of sildenafil on the Kv channel subfamily was examined via the special anti-Kv1.5 antibody dialysis. Real-time PCR and Western blotting techniques were performed to detect the gene transcription and protein expression of Kv1.5 channel in human PASMCs. Results Hypoxia obviously inhibited whole-cell K+ currents in PASMCs; sildenafil significantly restored hypoxia-decreased K+ currents; Kv1.5 channel is the target of the modulation of sildenafil on Kv in hypoxic PASMCs; sildenafil partially restored hypoxia-inhibited mRNA and protein expression of Kv1.5 channel in PASMCs. Conclusion Sildenafil inhibits hypoxia-induced voltage-dependent potassium channels downregulation in human PASMCs.

Key words: sildenafil, voltage-dependent potassium channels, human pulmonary artery smooth muscle cells

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