Department of Pharmacology, University of Saskatchewan, Saskatoon, Saskatchewan, Canada.
Proteomics. 2010 Dec;10(24):4377-85. doi: 10.1002/pmic.201000393. Epub 2010 Nov 17.
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.
人们越来越关注 Rho 激酶通路在心脏病和缺血/再灌注（I/R）损伤中的作用。Y-27632 是一种 Rho 激酶抑制剂，已被证明可预防 I/R 损伤，但确切的作用机制仍有待阐明。本项目的目的是确定 Y-27632 可以保护心脏免受 I/R 损伤的新靶点。在有氧条件下灌注分离的大鼠心脏，或在存在或不存在 Y-27632 的情况下进行 I/R。在缺血前和再灌注的前 10 分钟给予 Y-27632（1μM）可使心脏功能完全恢复。2-D 电泳后通过 MS 鉴定出四种受 Y-27632 处理影响的蛋白质。乳酸脱氢酶和甘油醛-3-磷酸脱氢酶在 Y-27632 处理组中显着增加，而肌酸激酶则恢复到对照水平。此外，我们发现 ATP 合酶的两种不同分子片段的水平增加，Y-27632 使其恢复正常。这种增加表明在缺血期间 ATP 合酶受到降解。代谢酶水平的变化及其受 Y-27632 的调节表明，Y-27632 的心脏保护作用涉及增加能量产生。
