title: Ischemia increases detectable endothelial nitric oxide synthase in rat and human myocardium.
abstract: The aims of the present study were to establish if myocardial ischemia/reperfusion is associated with altered eNOS activity and if myocardial eNOS detection depends on its activity. We determined detectable eNOS in (1) myocardium of isolated perfused rat hearts subjected to either global or regional ischemia and (2) in left ventricular biopsies from patients undergoing two different methods of myocardial protection (i.e., intermittent cold blood cardioplegia and continuous coronary perfusion with warm, beta-blocker-enriched blood) during coronary artery surgery. NOS detection was performed by NADPH-d staining and three eNOS-antibodies against different eNOS epitopes. In addition, activity dependent alteration of detectable eNOS was proofed by bradykinin treatment for 2 to 10 min. Ischemic and receptor mediated eNOS activation increased NADPH-d reactivity and eNOS immunoreaction as measured by antibodies against either amino acids of a central bovine eNOS domain or the human eNOS N-terminal end. In contrast, the antibody against the human eNOS C-terminal end exhibited no alteration of eNOS immunoreaction. The transient eNOS activation was associated with increased cGMP content. In human myocardium subjected to ischemia during cardiac surgery we found that early reperfusion increases eNOS activity. These data demonstrate a strong association between myocardial ischemia/reperfusion and increased eNOS activity as measured by immunocytochemical staining against specific eNOS epitopes. It appears that eNOS activation and subsequent NO release may act as a regulatory system to counter balance the potentially deleterious effects of myocardial ischemia/reperfusion.
authors.affiliation: Institut I for Anatomy, Clinic for Thoracic and Cardiovascular Surgery, University of Cologne, Joseph-Stelzmann-Strasse 9, Cologne, 50931, Germany. wilhelm.bloch@uni-koeln.de
authors.forename: W
authors.initials: W
authors.lastname: Bloch
authors.forename: U
authors.initials: U
authors.lastname: Mehlhorn
authors.forename: A
authors.initials: A
authors.lastname: Krahwinkel
authors.forename: M
authors.initials: M
authors.lastname: Reiner
authors.forename: M
authors.initials: M
authors.lastname: Dittrich
authors.forename: A
authors.initials: A
authors.lastname: Schmidt
authors.forename: K
authors.initials: K
authors.lastname: Addicks
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000818:Animals; D001920:Bradykinin / Q000032:analysis; D002314:Cardioplegic Solutions / Q000494:pharmacology; D004727:Endothelium / Q000201:enzymology; D004789:Enzyme Activation / Q000502:physiology; D006324:Heart Arrest, Induced; D006801:Humans; D007118:Immunoassay; D066298:In Vitro Techniques; D008297:Male; D017202:Myocardial Ischemia / Q000201:enzymology* / Q000473:pathology; D015425:Myocardial Reperfusion; D009206:Myocardium / Q000201:enzymology* / Q000473:pathology; D009252:NADPH Dehydrogenase / Q000032:analysis; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000319:Adrenergic beta-Antagonists; D002314:Cardioplegic Solutions; D019001:Nitric Oxide Synthase; D009252:NADPH Dehydrogenase; D001920:Bradykinin