Application of the nitric oxide donor SNAP to cardiomyocytes in culture provides protection against oxidative stress.
Multiple data indicates that nitric oxide (NO) donors retain immediate protective effects against different disturbances in cardiovascular system. The aim of the present study was to investigate delayed effects of nitric oxide donor S-nitroso-N-acetyl-l,l-penicillamine (SNAP) application in cardiac H9c2 cell line. Cardiomyocytes were treated with SNAP for 2h followed by 24h wash with fresh growth medium. The concentration curve was constructed in range from 0.5 to 2mM, toxicity was observed at 2mM concentration of SNAP. For the study of SNAP-induced protection against t-butyl hydroperoxide-induced oxidative injury 1mM concentration was used. Cell viability was assessed by MTT reductase activity assay; mitochondrial transmembrane potential (mdeltapsi) was measured by flow cytometry with fluorescent dye DiOC(6). Synthesis of heat-shock proteins (hsps) was analyzed by Western blot. Analysis of the cell viability and mdeltapsi reflected delayed protective effect of 1mM SNAP application against oxidative injury. SNAP in 1mM concentration caused 70% induction of hsp75 synthesis in cardiomyocytes. However, the other analyzed hsps (hsp70, hsp27, hsp60, hsp10, and CyP A) did not display any significant induction after incubation with SNAP. Present work demonstrates that the NO donor SNAP causes delayed protection against oxidative stress in H9c2 cardiomyocyte cell line, reflected in cell viability increase and preservation of the mdeltapsi. We suppose the major pathway for the development of SNAP-induced protection is through mitochondria. Induction of hsp75 expression following SNAP pretreatment is one possible way to explanation the mechanisms of this protection.
Monastyrskaya
Ekaterina
E
Institute of General Pathology and Pathophysiology, Baltijskaya 8, Moscow 125315, Russia.
Folarin
Najeem
N
Malyshev
Igor
I
Green
Colin
C
Andreeva
Larisa
L
D015153:Blotting, Western; D002460:Cell Line; D006360:Heat-Shock Proteins / Q000096:biosynthesis; D008564:Membrane Potentials / Q000187:drug effects; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism; D020030:Nitric Oxide Donors / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D010396:Penicillamine / Q000031:analogs & derivatives* / Q000494:pharmacology*
D006360:Heat-Shock Proteins; D020030:Nitric Oxide Donors; C110881:S-nitro-N-acetylpenicillamine; D009569:Nitric Oxide; D010396:Penicillamine