[1]杨明珠,陈依春,王晓露,等.钠-钙交换体单克隆抗体NCX-3F10对大鼠心室肌主要离子电流的影响及对缺血/再灌注诱发心律失常的抑制作用[J].中国药理学通报,2017,(07):934-941.[doi:10.3969/j.issn.1001-1978.2017.07.010]
 YANG Ming-zhu,CHEN Yi-chun,Wang Xiao-lu,et al.Investigation of effects of monoclonal antibody NCX-3F10 against Na+/Ca2+ exchanger on rat cardiac ionic currents and its suppression on ischemia-reperfusion induced cardiac arrhythmias[J].Chinese Pharmacological Bulletin,2017,(07):934-941.[doi:10.3969/j.issn.1001-1978.2017.07.010]
点击复制

钠-钙交换体单克隆抗体NCX-3F10对大鼠心室肌主要离子电流的影响及对缺血/再灌注诱发心律失常的抑制作用()
分享到:

《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2017年07期
页码:
934-941
栏目:
论著
出版日期:
2017-07-10

文章信息/Info

Title:
Investigation of effects of monoclonal antibody NCX-3F10 against Na+/Ca2+ exchanger on rat cardiac ionic currents and its suppression on ischemia-reperfusion induced cardiac arrhythmias
文章编号:
1001-1978(2017)07-0934-08
作者:
杨明珠1陈依春2王晓露1张堉琪1薛晓艳1封启龙1吴博威1
1. 山西医科大学生理学系,细胞生理学省部共建教育部重点实验室,山西 太原 030001;
2. 西藏民族大学医学部,陕西 咸阳 712000
Author(s):
YANG Ming-zhu1CHEN Yi-chun2Wang Xiao-lu1ZHANG Yu-qi1XUE Xiao-yan1FENG Qi-long1WU Bo-wei1
1. Dept of Physiology, Shanxi Medical University, Key Laboratory of Cellular Physiology of Shanxi Province and Ministry of Education, Taiyuan 030001,China;
2.Dept of Medicine, Tibet Nationalities University, Xianyang Shanxi 712000,China
关键词:
钠-钙交换体 单克隆抗体 全细胞膜片钳 离子电流 心律失常 缺血/再灌注
Keywords:
Na+-Ca2+ exchanger monoclonal antibody whole-cell patch clamp technique ion current cardiac arrhythmia ischemia-reperfusion
分类号:
R-332;R322.11;R331.38;R392.11;R541.7
DOI:
10.3969/j.issn.1001-1978.2017.07.010
文献标志码:
A
摘要:
目的 观察NCX-3F10抗体对大鼠心室肌细胞主要离子电流的影响及其对缺血/再灌注(ischemia/reperfusion, I/R)诱发心律失常的作用。方法 ① 利用全细胞膜片钳技术记录大鼠心室肌细胞Na+/Ca2+交换电流(INa/Ca)及其他主要离子电流; ② 结扎左冠状动脉建立大鼠在体及离体心脏缺血/再灌注心律失常模型,观察抗体对心律失常的影响; ③ 利用IonOptix离子影像分析系统观察抗体对单个心室肌细胞钙瞬变的影响。结果 ① 5~40 mg·L-1的NCX-3F10抗体对INa/Ca呈剂量依赖性抑制作用,其抑制外向和内向电流的IC50分别为11.15和11.69 mg·L-1,最大抑制率分别达到61%、62%; 该抗体对钙电流(ICa-L)也有一定的抑制作用,对内向整流钾电流(IK1)、瞬时外向钾电流(Ito)、钠电流(INa)均无影响; ② 在离体大鼠心脏I/R组,100%大鼠发生室速,88.89%大鼠出现室颤; 再灌注前5 min给予NCX-3F10抗体(10 mg·L-1)干预后,大鼠的室速发生率下降至44.43%,室速、室颤持续时间明显减少(P<0.05); ③ 麻醉大鼠再灌注前5 min给予NCX-3F10抗体(50 μg·kg-1)预处理后,大鼠室早个数、室速发生率及持续时间、室颤的发生率及持续时间均明显降低(P<0.05); ④ 5~40 mg·L-1的NCX-3F10抗体对心肌细胞钙瞬变幅度呈剂量依赖性抑制作用。结论 NCX-3F10抗体对大鼠离体和在体心脏缺血/再灌注诱发的心律失常有明显抑制作用,其抗心律失常效应与抑制钠-钙交换电流和L-型钙电流、减轻细胞内钙超载有关。
Abstract:
Aim To observe the effect of antibody NCX-3F10 on the main ion current of rat ventricular myocytes and its effect on arrhythmias induced by ischemia/reperfusion(I/R).Methods ① The whole-cell patch clamp technique was employed to record the Na+/Ca2+ exchange current(INa/Ca)and other major ion currents in rat ventricular myocytes.② The rat models of arrhythmia induced by ischemia/reperfusion were established by ligating the left coronary artery to in vivo and in vitro.Then the effects of antibody on the arrhythmia were observed.③ The IonOptix ion imaging system was used to observe the effect of antibody on calcium transients in single ventricular myocytes.Results ① The antibody NCX-3F10 dose-dependently inhibited INa/Ca from 5 to 40 mg·L-1. The IC50 for outward and inward currents was 11.15 and 11.69 mg·L-1, and the maximum inhibitory rates were 61% and 62%, respectively. The antibody also had an inhibitory effect on calcium current(ICa-L), and had no significant effect on inward rectifier potassium current(IK1), transient outward potassium current(Ito)and sodium current(INa). ② In the isolated rat heart group I/R, 100% rats showed ventricular tachycardia, and 88.89% rats had ventricular fibrillation. After administration of antibody NCX-3F10(10 mg·L-1)5 min before reperfusion, the incidence of ventricular tachycardia decreased to 44.43%(P<0.05), and the duration of ventricular tachycardia and ventricular fibrillation was also shortened remarkably(P<0.05). ③ In the anesthetized rats after administration of antibody NCX-3F10(50 μg·kg-1)5 min before reperfusion, the incidence and duration of ventricular tachycardia,the incidence and duration of ventricular fibrillation, and total number of ventricular premature beats were significantly decreased(P<0.05).④ From 5 to 40 mg·L-1, NCX-3F10 antibody decreased calcium transient amplitude in rat single ventricular myocytes dose-dependently(P<0.05).Conclusions The NCX-3F10 antibody shows significant arrhythmic effects on ischemia-reperfusion induced arrhythmia in rats both in vitro and in vivo, the underlying mechanism of which is related to NCX and L-type calcium current inhibition and calcium overload reduction by the NCX antibody.

参考文献/References:

[1] 隋 辉, 陈伟伟, 王 文.《中国心血管病报告2015》要点解读[J].中国心血管杂志, 2016, 21(4): 259-61. [1] Sui H, Chen W W, Wang W. Interpretation of report on cardiovascular diseases in China(2015)[J]. Chin J Cardiovasc Med,2016,21(4): 259-61.
[2] 王 文, 刘明波, 隋 辉,等. 中国心血管病的流行状况与防治对策[J]. 中国心血管病杂志, 2012, 17(5): 321-3.
[2] Wang W, Liu M B, Sui H, et al. The trend of cardiovascular disease in China and strategy of prevention[J]. Chin J Cardiovasc Med,2012,17(5): 321-3.
[3] Boscia F, D'Avanzo C, Pannaccione A, et al. Silencing or Knocking out the Na+/Ca2+ exchanger-3(NCX3)impairs oligodendrocyte differentiation[J]. Cell Death Differ. 2012, 19(4): 562-72.
[4] Parnis J, Montana V, Delgado-Martinez I, et al. Mitochondrial exchanger NCLX plays a major role in the intracellular Ca2+ signaling, gliotransmission, and proliferation of astrocytes[J]. J Neurosci,2013,33(17): 7206-19.
[5] Antoons G, Willems R, Sipido K R. Alternative strategies in arrhythmia therapy: evaluation of Na/Ca exchange as an anti-arrhythmic target[J]. Pharmacol Ther,2012,134(1): 26-42.
[6] Feng Q L, Wu D M, Cui X L, et al. Inhibitory effects of purified antibody against α-1 repeat(117-137)on Na+/Ca2+ exchange and L-type Ca2+ currents in rat cardio myocytes[J]. Acta Physiol Sin,2010,62(5): 407-14.
[7] Ottolia M, Nicoll D A,Philipson K D. Mutational analysis of the alpha-1 repeat of the cardiac Na+-Ca2+ exchanger[J].J Biol Chem,2005,280(2):1061-9.
[8] Shigekawa M, Iwamoto T, Uehara A,Kita S. Probing ion binding sites in the Na+/Ca2+ exchanger[J].Ann NY Acad Sci,2002,976:19-30.
[9] Schwarz E M,Benzer S. Calx, a Na-Ca exchanger gene of Drosophila melanogaster[J].Proc Natl Acad Sci USA,1997,94(19): 10249-54.
[10] 王 波.钠钙交换体对心肌保护作用的研究进展[J]. 西安文理学院学报:自然科学版, 2009, 12(3): 46-8.
[10] Wang B. New research progress of sodium/calcium exchanger in cardio protection[J]. J Xi'an Univer Arts Sci(Nat Sci Md), 2009, 12(3): 46-8.
[11] 邵 莹,吴启南,周 婧,等. 淡竹叶黄酮对大鼠心肌缺血/再灌注损伤的保护作用[J]. 中国药理学通报, 2013, 29(2): 241-7.
[11] Shao Y, Wu Q N, Zhou J, et al. Protective effects of total flavones from Lophatherum gracile on myocardial ischemia-reperfusion injury in rats[J]. Chin Pharmacol Bull, 2013, 29(2): 241-7.
[12] 孙 瑶, 张 婵, 简 洁. 九龙藤总黄酮调控自噬对抗心肌缺血/再灌注损伤的实验研究[J]. 中国药理学通报, 2015, 31(2): 232-6.
[12] Sun Y, Zhang C, Jian J. Effects of Bauhinia championii flavones on adjusting autophagy against myocardial ischemia/reperfusion injury[J]. Chin Pharmacol Bull, 2015, 31(2): 232-6.
[13] 贺永贵, 李王芳, 尹红丽,等. 黄芪甲苷抑制 GSK-3β 活性介导大鼠心肌缺血/再灌注损伤作用的线粒体机制研究[J]. 中国药理学通报, 2014, 30(3): 402-6.
[13] He Y G, Li W F, Yin H L, et al. Mitochondrial mechanism of Astragaloside IV induced inhibition of GSK-3β in myocardial ischemia/reperfusion injury in rats[J]. Chin Pharmacol Bull, 2014, 30(3): 402-6.
[14] Goldhaber J I. Philipson K D. Cardiac sodium-calcium exchange and efficient excitation-contraction coupling: implications for heart disease[J]. Adv Exp Med Biol, 2013, 961:355-64.
[15] Nicoll D A, Ottolia M, Lu L, et al. A new topological model of the cardiac sarcolemmal Na+-Ca2+ exchanger[J].J Biol Chem,1999, 274(2): 910-7.
[16] Ren X, Philipson K D. The topology of the cardiac Na+/Ca2+ exchanger, NCX1[J].J Mol Cell Cardiol,2013,57:68-71.
[17] Szerencsei R T, Kinjo T G, Schnetkamp P P. The topology of the C-terminal sections of the NCX1 Na(+)/Ca(2+)exchanger and the NCKX2 Na(+)/Ca(2+)-K(+)exchanger[J].Channels,2013,7(2):109-14.

相似文献/References:

[1]杨啸燕,李明华,赵雷,等.抗钠-钙交换体α1(106145)抗体对大鼠离体心功能的影响及其机制[J].中国药理学通报,2012,(08):1074.
 YANG Xiao yan,LI Ming hua,ZHAO Lei,et al.Effects of antibody against α1(106145) on cardiac function in isolated rat hearts[J].Chinese Pharmacological Bulletin,2012,(07):1074.
[2]陈雅慧,刘 冬,彭礼飞.凝血因子XI作为抗栓防治新靶点的研究进展[J].中国药理学通报,2015,(05):619.
 CHEN Ya-hui,LIU Dong,PENG Li-fei.Research progress on factor XI as a novel target for antithrombotic therapy[J].Chinese Pharmacological Bulletin,2015,(07):619.
[3]方 蕾,侯 睿,费巧玲,等.虾原肌球蛋白及其IgE单克隆抗体建立I型过敏反应体内外模型[J].中国药理学通报,2016,(10):1422.[doi:10.3969/j.issn.1001-1978.2016.10.018]
 FANG Lei,HOU Rui,et al.The in vitro and in vivo models induced by shrimp tropomyosin and monoclonal tropomyosin-specific murine IgE antibody[J].Chinese Pharmacological Bulletin,2016,(07):1422.[doi:10.3969/j.issn.1001-1978.2016.10.018]
[4]薛晓艳,王苗苗,王晓露,等.钠-钙交换体单克隆抗体NCX-2D2对异丙肾上腺素诱发成年大鼠心律失常的抑制作用[J].中国药理学通报,2018,(09):1308.[doi:10.3969/j.issn.1001-1978.2018.09.023]
 XUE Xiao-yan,WANG Miao-miao,WANG Xiao-lu,et al.Inhibitory effect of monoclonal antibody NCX-2D2 on isoproterenol-induced arrhythmias in adult rat hearts[J].Chinese Pharmacological Bulletin,2018,(07):1308.[doi:10.3969/j.issn.1001-1978.2018.09.023]

备注/Memo

备注/Memo:
基金项目:山西省重点实验室建设项目(No 2015012003-08); 山西省高等学校优秀青年学术带头人支持项目; 高校“131”领军人才工程优秀中青年拔尖创新人才
作者简介:杨明珠(1990-),女,硕士生,研究方向:心肌电生理,E-mail: 15386745259@163.com;
封启龙(1976-),男,博士,硕士生导师,研究方向:心肌电生理,通讯作者,Tel:0351-4135768, E-mail: fengqilong@163.com
更新日期/Last Update: 2017-07-15