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Amiloride和Ni~(2+)抑制缺血/复灌引起的大鼠心肌细胞内钙的增力

董建文，朱海峰，周兆年

中国科学院上海生命科学研究院低氧心血管生理实验室，上海200031

摘要

旨在研究Na`/H+交换以及Na~(+) / Ca~(2+)交换对模拟缺血/复灌引起的大鼠心肌细胞内游离钙水平变化的调节作用。分别利用模拟缺血液和正常台氏液对大鼠心肌细胞进行缺血/复灌处理，在缺血期间分别应用Na`/H+交换抑制剂阿米洛利(a mil oride) . Na` / Ca~(2+)交换抑制剂N~(+)Ca~(2)以及无钙液，观察它们对细胞内游离Ca~(2+)浓度变化的影响。利用Zeiss- LS M 51。激光共聚焦显微镜检测采集细胞内游离ca~(2+)的指示剂F1uo 3- AM的荧光信号，计算出相对于正常(缺血前)的相对荧光强度，以表示胞内游离"2+浓度的变化。结果显示，模拟缺血引起大鼠心肌细胞内游离。2+持续上升.

关键词： 心肌细胞; 缺血再灌注; 钙离子; Na~(+)/H~(+)交换; Na~(+)` / Ca~(2+)交换

Inhibitors of Na~(+)/ H~(+)and Na ~(+)/ Ca~(2+)exchange depress intracellular calcium elevation induced by ischemia/reperf union in rat cardiacmyocytes

DONG Jian Wen

aboratory of Hypoxic Cardiovascular Physiology，Shanghai Institutes for Biological Science ，Chinese Academy of Science，Shanghai

Abstract

An increase in cytosolic free calcium has been shown to occur during ischemia in perfused hearts and plays a pivotal role in ischemia/ reperfusion injury.The objective‘this study was to investigate the contributions‘Na`/H `exchange and M~(+)/cat+exchange to changes in intracellular calcium([Ca~(2+)];) during simulated ischemia and reperfusion in quiescent isolated rat cardiac myocytes.[Ca 2+]; was measured by laser confocal microscope using the fluorescent indicator Huo 3 and expressed as the corrected intensity‘Huo 3 fluorescence.[ca~(2+)]1 increased to 140. 3士13.0%(P(0.05preischemic control 100%)after 5 min simulated isthe mia，and remained at high level of 142.8士15.5%(P(0. 05) after the following 15 min reperfusion.The increase in[ca~(2+)]1 during simulated ischemia and reperfusion was suppressed by 100 umol/ L amiloride(inhibitor of M~(+)/H` exchanger)，5 mmol/ L NiCl2(inhibitor of Na~(+)/cat+exchanger) and calcium-free solution;[cat+]1 was 101.4士16. 3%，110.4士11.1%and 107.1士10.8%，respectively，after 5 min simulated ischemia，and 97.8士14.3%，106.2士14.5%and 106.6士15.7%，respectively，after 15 minre on .Compared with control cells，the amplitude of spontaneous calcium oscillation was lessened in cells treated with(rfree perfusion and MCl2 during reperfusion .In addition，no calcium oscillation was observed in cells pretreated with .These results suggest that MH+exchange and Na ca~(2+)exchange are activated during simulated ischemia in isolated quiescent cardiac myocytes，leading to the elevation of[Ca~(2+)]; induced by simulated ischemia and reperfusion.

Key words: cardiac myocytes;ischemia/ reperfusion;Calcium;Na+/H+exchanger;sodium- calcium exchanger

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引用本文：

董建文，朱海峰，周兆年. Amiloride和Ni~(2+)抑制缺血/复灌引起的大鼠心肌细胞内钙的增力[J]. 生理学报 2003; 55 (3): .

DONG Jian Wen. Inhibitors of Na~(+)/ H~(+)and Na ~(+)/ Ca~(2+)exchange depress intracellular calcium elevation induced by ischemia/reperf union in rat cardiacmyocytes . Acta Physiol Sin 2003; 55 (3): (in Chinese with English abstract).