Effect of the Rho kinase inhibitor Y-27632 on the proteome of hearts with ischemia-reperfusion injury.
Growing attention has been given to the role of the Rho kinase pathway in the development of heart disease and ischemia/reperfusion (I/R) injury. Y-27632 is a Rho kinase inhibitor demonstrated to protect against I/R injury, but the exact mechanism by which it does so remains to be elucidated. The goal of this project was to determine new targets by which Y-27632 can protect the heart against I/R injury. Isolated rat hearts were perfused under aerobic conditions or subjected to I/R in the presence or absence of Y-27632. Administration of Y-27632 (1 μM) before ischemia and during the first 10 min of reperfusion resulted in complete recovery of cardiac function. 2-D electrophoresis followed by MS identified four proteins whose levels were affected by Y-27632 treatment. Lactate dehydrogenase and glyceraldehyde-3-phosphate dehydrogenase were significantly increased in the Y-27632 treated group, while creatine kinase was normalized to control levels. In addition, we found increased level of two different molecular fragments of ATP synthase, which were normalized by Y-27632. This increase suggests that during ischemia ATP synthase is subjected to degradation. The changes in metabolic enzymes' levels and their regulation by Y-27632 suggest that the cardioprotective effect of Y-27632 involves increased energy production.
Department of Pharmacology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Virgilio J J
VJ
Cadete
Jolanta
J
Sawicka
Dorota
D
Polewicz
Adrian
A
Doroszko
Mieczyslaw
M
Wozniak
Grzegorz
G
Sawicki
D000577:Amides / Q000494:pharmacology*; D000595:Amino Acid Sequence; D000818:Animals; D008297:Male; D025261:Mitochondrial Proton-Translocating ATPases / Q000378:metabolism; D008969:Molecular Sequence Data; D009200:Myocardial Contraction / Q000187:drug effects; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000503:physiopathology; D009206:Myocardium / Q000378:metabolism*; D047428:Protein Kinase Inhibitors; D021122:Protein Subunits / Q000378:metabolism; D020543:Proteome / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
D000577:Amides; D047428:Protein Kinase Inhibitors; D021122:Protein Subunits; D020543:Proteome; D011725:Pyridines; C108830:Y 27632; D054460:rho-Associated Kinases; D025261:Mitochondrial Proton-Translocating ATPases