| 肺动脉平滑肌细胞钾电流记录方法的建立 |
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| 引用本文： | 邝素娟,钱卫民,耿庆山,邓春玉,单志新,林秋雄,杨敏,余细勇,吴书林,林曙光. 肺动脉平滑肌细胞钾电流记录方法的建立[J]. 岭南心血管病杂志, 2009, 15(4): 318-321. DOI: 10.3969/j.issn.1007-9688.2009.04.021 |
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| 作者姓名： | 邝素娟 钱卫民 耿庆山 邓春玉 单志新 林秋雄 杨敏 余细勇 吴书林 林曙光 |
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| 作者单位： | 广东省人民医院(医学研究中心),广东省医学科学院,广东省心血管病研究所,广州,5100100 |
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| 基金项目： | 广东省医学科研基金资助项目，广东省科技计划资助项目(2006836002023)国家自然科学基金资助项目 |
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| 摘 要： | 目的 在肺动脉平滑肌细胞(pulmonary arterial smooth muscle cells,PASMCs)上建立钙离子激活钾通道(calcium-activated potassium channel,KCa)电流、电压门控钾通道(voltage-gated potassium channel,Ky)电流和内向整流钾通道(Inward rectifier channel,Ku)电流的记录方法 .方法 用急性酶解的方法 分离出单个大鼠PASMC,利用全细胞膜片钳方法 记录钾电流.结果 ①急性酶解分离得到高质量的单个大鼠PASMC,呈梭形,边界清楚,胞浆均匀透亮.②Kv电流可以被5 mmol/L4-AP明显抑制,使电流-电压关系曲线明显下移,在55 mV时,Kv电流密度从(133.86±7.36)pA/pF减少到(59.09±3.35)pA/pF(n=6,P<0.05).③1 mmol/L TEA对KCa电流有明显抑制作用,使电流-电压关系曲线明显下移,在55 mV时,KCa电流密度从(9.03±1.42)pA/pF减少到(2.12±0.52)pA/pF(n=7,P<0.05).④KATP电流可以被10 μmol/L尼可地尔激活,使电流-电压关系曲线明显上移,在50 mV时,Kv电流密度从(29.08±5.90)pA/pF增加到(88.90±7.98)pA/pF(n=10,P<0.05).结论 在肺动脉平滑肌细胞上成功地建立了KCa、Kv和Kir电流的记录方法 ,可以为肺动脉相关疾病的发病机制和治疗方案的探索,提供有效的细胞模型基础.
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| 关 键 词： | 肺动脉 肌细胞,平滑肌 钾 钙 |
Establishment of recording potassium channel currents in pulmonary arterial smooth muscle cells |
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| KUANG Su-juan,QIAN Wei-min,GENG Qing-shan,DENG Chun-yu,CHAN Zhi-xin,MN Qiu-xiong,YANG Min,YU Xi-yong,WU Shu-lin,LIN Shu-guang. Establishment of recording potassium channel currents in pulmonary arterial smooth muscle cells[J]. South China Journal of Cardiovascular Diseases, 2009, 15(4): 318-321. DOI: 10.3969/j.issn.1007-9688.2009.04.021 |
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| Authors: | KUANG Su-juan QIAN Wei-min GENG Qing-shan DENG Chun-yu CHAN Zhi-xin MN Qiu-xiong YANG Min YU Xi-yong WU Shu-lin LIN Shu-guang |
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| Affiliation: | Medical Research Center of Guangdong General Hospital;Guangdong Cardiovascular Institute;Guangdong Academy of Medical Sciences;Guangzhou 510080 |
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| Abstract: | Objectives To establish the methods for recording calcium-activated potassium channel ( KCa) , voltage-gated potassium channel (KV) and inward rectifier channel (Kir) in pulmonary arterial smooth muscle cells (PASMC). Methods PASMC was acutely isolated by enzyme. Potassium currents were recorded by whole-cell patch- clamp technique. Results (1) High-quality single PASMCs were obtained. A typical single cell was fusiform shape and its cytoplasm was uniform and lucent. (2) KV currents were inhibited by 5 mmol... |
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| Keywords: | pulmonary artery smooth muscle cell potassiun calcium |
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