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|本期目录/Table of Contents|

人心肌细胞缓慢激活延迟整流钾电流细胞模型的建立(PDF)

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2007年第1期
页码:
28-31/35
栏目:
基础研究
出版日期:
2007-01-01

文章信息/Info

Title:
Establishment of cell model of human cardiomyocyte slowly activating delayed rectifier potassium current
作者:
孙冬冬1; 王晓斌2; 赵志敬1; 李志超2; 朱妙章3; 贾国良1; 王海昌1; 董明清2
第四军医大学：1.西京医院心血管内科，2.基础部病理生理学教研室，3.基础部生理学教研室，陕西 西安 710032
Author(s):
SUN Dongdong1;  WANG Xiaobin2;  ZHAO Zhijing1; LI Zhichao2;  ZHU Miaozhang3;  JIA Guoliang1;  WANG Haichang1;  DONG Mingqing2
1.Department of Cardiology, Xijing Hospital, 2.Department of Pathophysiology, 3.Department of Physiology, Fourth Military Medical University, Xi′an 710032,Shaanxi, China
关键词:
缓慢激活延迟整流钾电流; KCNQ1/KCNE1; HEK 293细胞系
Keywords:
slowly activating delayed rectifier potassium current; KCNQ1/KCNE1;  HEK 293 cell line
分类号:
R331.38
DOI:
-
文献标识码:
A
摘要:
目的 建立稳定表达人心肌细胞缓慢激活延迟整流钾电流（IKs）的细胞模型。方法 编码IKs通道α亚单位的KCNQ1基因及β亚单位的KCNE1基因共转染HEK 293细胞，潮霉素B筛选，电生理学及药理学方法鉴定。结果 KCNQ1/KCNE1基因被成功转入HEK 293细胞，KCNQ1/KCNE1电流与人心肌IKs电流具有相似的电流特性；hKCNQ1/hKCNE1通道反转电位与细胞外钾离子浓度呈线性关系；选择性IKs通道阻断剂Chromanol 293B对KCNQ1/KCNE1电流具有明显而可逆的抑制作用，其IC50 (+40 mV)为9.1 μmol/L；无钾细胞外液可以增加KCNQ1/KCNE1电流幅度，+40 mV时电流幅度增加(28.6±2.0)%（P<0.01, n=8），但对其动力学特性无明显影响。结论 已经成功构建稳定表达人心肌KCNQ1/KCNE1通道蛋白的HEK 293细胞系，其电生理学特性和药理学特性与人心肌IKs相似，可以作为研究人心肌IKs的细胞模型。
Abstract:
AIM To establish a cell model of human cardiomyocyte delayed rectifier potassium current (IKs). METHODS Human cardiac KCNQ1 and KCNE1 genes, which encode α and β subunit of IKs respectively, were cotransfected into HEK 293 cells. Hygromycin B was used to screen the transfected cells. Electrophysiological method was used to identify stably expressing KCNQ1/KCNE1 HEK 293 cell clones. RESULTS KCNQ1/KCNE1 genes were successfully transfected into HEK 293 cells. The KCNQ1/KCNE1 currents had similar characteristics of native human IKs. Its reverse potential was linearly correlated with the extracellular potassium ions. The currents were significantly and reversibly inhibited by Chromanol 293B, a specific IKs blocker, with an IC50 of 9.1 μmol/L. Extracellular fluid without potassium ions significantly increased the magnitude of IKs but had no effects on its kinetics. At +40 mV, the increased level was (28.6±2.0)% (P<0.01, n=8). CONCLUSION HEK 293 cell model stably expressing human cardiac KCNQ1/KCNE1 channels has been successfully established, which has similar characteristics of native human cardiac IKs. The established cell model is an ideal cell model of human cardiac IKs.

参考文献/References

［1］Li GR,Feng J,Yue L,et al.Evidence for two components of delayed rectifier K+ current in human ventricular myocytes［J］. Circ Res, 1996, 78(5): 689-696.

［2］Sanguinetti MC, Curran ME, Zou A,et al. Coassembly of K(V)LQT1 and minK (IsK) proteins to form cardiac I(Ks) potassium channel［J］. Nature, 1996, 384(6604): 80-83.

［3］Chen H, Kim LA, Rajan S,et al. Charybdotoxin binding in the I(Ks) pore demonstrates two MinK subunits in each channel complex［J］. Neuron, 2003, 40(1): 15-23.

［4］Sanguinetti MC. Dysfunction of delayed rectifier potassiumm channels in an inherited cardiac arrhythmia［J］. Ann N Y Acad Sci,1999,868:406-413.

［5］ Li GR,Lau CP,Ducharme A,et al. Transmural action potential and ionic current remodeling in ventricles of failing canine hearts［J］. Am J Physiol Heart Circ Physiol,2002,283(3): H1031-H1041.

［6］Li GR,Lau CP,Leung TK,et al.Ionic current abnormalities associated with prolonged action potentials in cardiomyocytes from diseased human right ventricles［J］.Heart Rhythm,2004,1(4): 460-468.

［7］Xu X,Rials SJ,Wu Y,et al. Left ventricular hypertrophy decreases slowly but not rapidly activating delayed rectifier potassium currents of epicardial and endocardial myocytes in rabbits［J］. Circulation, 2001, 103(11):1585-1590.

［8］Seebohm G,Lerxhe C,Pusch M,et al. A kinetic study on the stereospecific inhibition of KCNQ1 and I(Ks) by the chromanol 293B［J］. Br J Pharmacol, 2001,134(8):1647-1654.

［9］Bosch RF, Gaspo R, Busch AE,et al. Effects of the chromanol 293B, a selective blocker of the slow, component of the delayed rectifier K+ current, on repolarization in human and guinea pig ventricular myocytes［J］. Cardiovasc Res,1998,38(2):441-450.

［10］Volders PG, Stengl M, van Opstal JM,et al. Probing the contribution of IKs to canine ventricular repolarization: Key role for betaadrenergic receptor stimulation［J］. Circulation,2003,107(21):2753-2760.

［11］Seebohm G, Lerche C, Busch AE, et al.Dependence of I(Ks) biophysical properties on the expression system［J］. Pflugers Arch, 2001, 442(6): 891-895.

［12］Sanguinetti MC, Jurkiewicz NK.Role of external Ca2+ and K+ in gating of cardiac delayed rectifier K+ currents［J］. Pflugers Arch, 1992, 420(2): 180-186.

备注/Memo

备注/Memo:
收稿日期：2006-4-13.基金项目：国家自然科学基金项目资助（No.30400155） 通讯作者：董明清，讲师，从事心血管离子通道调控的研究Email:dongmqxh@fmmu.edu.cn 共同通讯作者：王海昌，主任医师，主要从事心脏疾病的基础与临床研究Email:wanghc@fmmu.edu.cn 作者简介：孙冬冬，硕士生Email:sundd3@fmmu.edu.cn
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