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中国应用生理学杂志 ›› 2018, Vol. 34 ›› Issue (3): 218-222.doi: 10.12047/j.cjap.5581.2018.052

• 研究论文 • 上一篇    下一篇

细胞外Ba2+对大鼠心肌细胞L型钙通道的影响

孟红旭, 姚明江, 任钧国, 刘建勋   

  1. 中国中医科学院西苑医院基础医学研究所, 中药药理北京市重点实验室, 北京, 100091
  • 收稿日期:2017-03-09 修回日期:2017-11-29 出版日期:2018-05-28 发布日期:2018-09-08
  • 通讯作者: 刘建勋,Tel:010-62835601;E-mail:liujx0324@sina.com E-mail:liujx0324@sina.com
  • 基金资助:
    中国中医科学院自助选题项目(zz070821);973计划课题(2015CB554405);中国中医科学院创新团队建设项目(YS1303)

Effects of extracellular Ba2+ on L-type calcium current in rat ventricular cardiocytes

MENG Hong-xu, YAO Ming-jiang, REN Jun-guo, LIU Jian-xun   

  1. Institute of Basic Medical Sciences of Xiyuan Hospital, China Academy of Chinese Medical Sciences, Beijing key Laboratory of Pharmacology of Chinese Materia Medica, Beijing 100091, China
  • Received:2017-03-09 Revised:2017-11-29 Online:2018-05-28 Published:2018-09-08
  • Supported by:
    中国中医科学院自助选题项目(zz070821);973计划课题(2015CB554405);中国中医科学院创新团队建设项目(YS1303)

摘要: 目的:观察细胞外Ba2+对记录大鼠心肌细胞L型钙通道的影响。方法:采用急性酶解分离法获得大鼠的单个心肌细胞,使用全细胞膜片钳技术记录L型钙通道电流。采用Ba2+替换台式液中的Ca2+和直接向台式液中加入Ba2+(0~8 mmol/L),观察峰值电流15 min内的变化,数据采用5个以上细胞进行重复。结果:(1)台式液中的Ca2+被Ba2+替换后,L型钙通道电流的失活速率明显减慢(P<0.01);在台式液中加入少量Ba2+(0.2,0.4 mmol/L)时L型钙通道电流的失活速率无明显改变(P>0.05),加入0.8 mmol/L Ba2+时失活速率明显减慢(P<0.05)。(2)与正常台式液比较,在细胞外液中加入Ba2+(0.2,0.4 mmol/L)峰值电流衰减减弱,其中10 min和15 min两个时间点衰减差异明显(P<0.01)。(3)在细胞外液中加入Ba2+可下移电流电压曲线,改变翻转电位,减弱丹酚酸A对钙电流的抑制强度,使量效关系曲线右移。结论:在细胞外液中加入一定浓度的Ba2+,能够减弱全细胞膜片钳技术记录大鼠心室肌细胞L型钙通道时出现的峰值电流衰减,改变通道的电压依赖特性,影响药物量效关系。

关键词: 大鼠, 心肌细胞, Ba2+, L型钙通道, 全细胞膜片钳

Abstract: Objective: To observe the effect of extracellular Ba2+ on the recording L-type calcium channel of rat ventricular cardiocytes.Methods: Whole-cell patch clamp technique was used to record the currents of L-type calcium channels (ICa,L) in a single ventricular cardiocyte isolated from a rat heart by acute enzymatic digestion combined with mechanical method. Ba2+ was used to replace Ca2+ in Tyrode's solution, and Ba2+ (0~0.8 mmol/L) was directly added to Tyrode's solution to observe the change of the peak current within 15 minutes. The data were repeated with 5 cells or more.Results: ①The inactivation rate of ICa,L was slowed down significantly (P<0.01) with Ba2+ instead of Ca2+ gradually in Tyrode's solution. There was no change in the inactivation rate of ICa,L when the Ba2+ (0.2, 0.4 mmol/L) was added in the extracellular solution, but when 0.8 mmol/L Ba2+ was added, the inactivation rate would slow down significantly (P<0.01). ②Ba2+(0.2, 0.4 mmol/L) added in the extracellular solution could decrease the peak current attenuation. Compared with the group in Tyrode's solution, the peak current attenuation rate of the group in Ba2+ (0.2, 0.4 mmol/L) added in the extracellular solution was decreased significantly at the two time points (10 min, 15 min, P<0.01). ③The extracellular Ba2+ (0.4 mmol/L) could move the current-voltage curve down, change the reversal potential, weaken ICa,L inhibition induced by salvianolic acid A and right-shift concentration-response curve.Conclusion: The addition of Ba2+ to extracellular solution can attenuate the peak current decay, change the voltage-dependent characteristics of calcium channels and influence drug dose-effect relationship when whole-cell patch clamp technique is used to record L-type calcium channel of rat ventricular cardiocytes.

Key words: rat, ventricular myocytes, Ba2+, L-type calcium current, whole-cell patch clamp

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