Ultrastructural study of calcium shift in ischemic/reperfused rat heart under treatment with dimethylthiourea, diltiazem and amiloride.
Among factors underlying reperfusion injury are oxygen free radicals and Ca2+ influx via gated calcium channel or via Na+/H(+)-Na+/Ca2+ exchange which lead to calcium overload. The aim of the study was to ultrastructurally visualize the distribution of Ca2+ and to compare binding of calcium by the sarcolemma and calcium accumulation in mitochondria under therapy with an OH scavenger, dimethylthiourea (DMTU), Na+/H+ exchange inhibitor, amiloride, and calcium channel blocker, diltiazem, given alone or in combination to ischemic/reperfused hearts. Isolated working hearts subjected to 40 min ischemia and 30 min reperfusion were perfused with drugs added to the perfusate 15 min before ischemia and administered for the rest of the perfusion period. The cytochemical phosphate pyroantimonate method for localization of Ca2+ was used, and calcium distribution was analyzed with a computer image analyzer. All drugs given alone improved sarcolemmal ability to bind calcium. The best results were obtained with amiloride. All of the combined therapies gave even better results, but calcium accumulation in mitochondria diminished only with diltiazem therapy given alone or in combination with DMTU. Since the presence of Ca2+ deposits on the sarcolemma is believed to represent its normal function, and calcium sequestration by mitochondria reflects an increase in cytosolic calcium load, the lack of correlation between sarcolemmal and mitochondrial Ca2+ distribution might suggest impaired mechanisms of lowering cytoplasmic calcium or the existence of some mechanism other than Na+/Ca2+ exchange, mediated by activated Na+/H+ exchange.
Department of Pathology, Children's Memorial Health Institute, Warsaw, Poland.
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Czarnowska
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Karwatowska-Prokopczuk
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Kurzydlowski
D000584:Amiloride / Q000627:therapeutic use*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000627:therapeutic use*; D004110:Diltiazem / Q000627:therapeutic use*; D004359:Drug Therapy, Combination; D016166:Free Radical Scavengers / Q000627:therapeutic use*; D006321:Heart / Q000187:drug effects; D017665:Hydroxyl Radical / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008929:Mitochondria, Heart / Q000378:metabolism; D017202:Myocardial Ischemia / Q000378:metabolism*; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D009206:Myocardium / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D012508:Sarcolemma / Q000378:metabolism; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors*; D013890:Thiourea / Q000031:analogs & derivatives* / Q000627:therapeutic use
D002121:Calcium Channel Blockers; D016166:Free Radical Scavengers; D017923:Sodium-Hydrogen Exchangers; D017665:Hydroxyl Radical; D000584:Amiloride; C038983:1,3-dimethylthiourea; D004110:Diltiazem; D013890:Thiourea; D002118:Calcium