Makino N, Panagia V, Gupta M P, Dhalla N S
Division of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, Winnipeg, Canada.
Circ Res. 1988 Aug;63(2):313-21. doi: 10.1161/01.res.63.2.313.
Na+-Ca2+ exchange and Ca2+-pump activities were studied in sarcolemmal vesicles isolated from rat hearts subjected to "calcium paradox" on perfusion with Ca2+-free medium followed by reperfusion with medium containing 1.25 mM Ca2+. Perfusion of hearts with Ca2+-free medium for 5 minutes did not affect the Na+-dependent Ca2+ uptake, ATP-dependent Ca2+ uptake, or Ca2+-stimulated ATPase activities in sarcolemma. Reperfusion of the Ca2+-deprived hearts with medium containing Ca2+ for 1-2 minutes increased Na+-dependent Ca2+ uptake, whereas reperfusion for 5-10 minutes decreased Na+-dependent Ca2+ uptake in sarcolemmal vesicles. Both ATP-dependent Ca2+ uptake and Ca2+-stimulated ATPase activities in sarcolemma were depressed on reperfusion of Ca2+-deprived hearts for 2-10 minutes. Reperfusion of Ca2+-deprived hearts for 5 minutes, which failed to generate contractile force, resulted in contracture without any recovery of the contractile force development. These changes in sarcolemmal Ca2+ transport and contractile function were prevented when hearts were perfused with Ca2+-free medium either in the presence of low sodium (35 mM) or at a low temperature (21 degrees C) before starting the reperfusion. No alterations in the purity of the preparation or permeability of sarcolemmal vesicles with respect to Na+ or Ca2+ were detected in hearts perfused with Ca2+-free medium or on reperfusion with medium containing calcium. The results indicate abnormalities in sarcolemmal Na+-Ca2+ exchange and Ca2+-pump mechanisms on reperfusion of Ca2+-deprived hearts with medium containing Ca2+, and such changes may partly account for the occurrence of intracellular Ca2+ overload during the development of calcium paradox.
研究了从经历“钙反常”的大鼠心脏分离的肌膜囊泡中的钠钙交换和钙泵活性。在无钙培养基中灌注5分钟后,再用含1.25 mM钙的培养基进行再灌注。用无钙培养基灌注心脏5分钟,对肌膜中钠依赖性钙摄取、ATP依赖性钙摄取或钙刺激的ATP酶活性没有影响。用含钙培养基对缺钙心脏进行1 - 2分钟的再灌注增加了钠依赖性钙摄取,而5 - 10分钟的再灌注则降低了肌膜囊泡中钠依赖性钙摄取。在对缺钙心脏进行2 - 10分钟的再灌注后,肌膜中ATP依赖性钙摄取和钙刺激的ATP酶活性均降低。对缺钙心脏进行5分钟的再灌注未能产生收缩力,导致挛缩,且收缩力未恢复。当在再灌注开始前,心脏在低钠(35 mM)存在下或低温(21摄氏度)下用无钙培养基灌注时,肌膜钙转运和收缩功能的这些变化得以预防。在用无钙培养基灌注的心脏或用含钙培养基再灌注的心脏中,未检测到制剂纯度或肌膜囊泡对钠或钙的通透性有改变。结果表明,在用含钙培养基对缺钙心脏进行再灌注时,肌膜钠钙交换和钙泵机制存在异常,这些变化可能部分解释了钙反常发生过程中细胞内钙超载的出现。