背景:特发性扩张型心肌病是最常见的心肌病类型之一。有人提出,心力衰竭中氧化应激的增加导致一氧化氮信号的减少,导致亚硝基氧化还原信号受损。为了检验这个假设,我们调查了来自外植扩张型心肌病和未衰竭供体男性和女性人类心脏的活检中蛋白质S-亚硝基化(SNO)和氧化的发生情况.
结果:氧化还原基树脂辅助捕获,用于氧化和SNO蛋白质组学分析,用于测量蛋白质氧化和SNO,分别。此外,使用马来酰亚胺巯基反应性荧光剂的二维差异凝胶电泳用于鉴定SNO蛋白。与健康供体相比,扩张型心肌病活检组织中的蛋白质氧化增加。有趣的是,在衰竭的心脏中,我们没有发现SNO持续下降;我们发现一些蛋白质显示SNO增加,而另一些则显示减少,反应存在性别差异。在女性心脏衰竭中,我们发现了10种SNO显着减少的蛋白质和4种SNO增加的蛋白质。比较未衰竭和衰竭的男性心脏,我们发现9种蛋白质显着减少,12种蛋白质显着增加。我们还发现,在衰竭的女性和男性心脏中,内皮一氧化氮合酶的S-谷胱甘肽化增加,表明女性心脏中的非偶联一氧化氮合酶增加。
结论:这些发现强调了亚硝基氧化还原信号在生理和病理条件下的重要性,提示治疗心力衰竭的潜在目标。
BACKGROUND: Idiopathic dilated cardiomyopathy is one of the most common types of cardiomyopathy. It has been proposed that an increase in oxidative stress in heart failure leads to a decrease in nitric oxide signaling, leading to impaired nitroso-redox signaling. To test this hypothesis, we investigated the occurrence of protein S-nitrosylation (SNO) and oxidation in biopsies from explanted dilated cardiomyopathy and nonfailing donor male and female human hearts.
RESULTS: Redox-based resin-assisted capture for oxidation and SNO proteomic analysis was used to measure protein oxidation and SNO, respectively. In addition, 2-dimensional difference gel electrophoresis using maleimide sulfhydryl-reactive fluors was used to identify the SNO proteins. Protein oxidation increased in dilated cardiomyopathy biopsies in comparison with those from healthy donors. Interestingly, we did not find a consistent decrease in SNO in failing hearts; we found that some proteins showed an increase in SNO and others showed a decrease, and there were sex-specific differences in the response. We found 10 proteins with a significant decrease in SNO and 4 proteins with an increase in SNO in failing female hearts. Comparing nonfailing and failing male hearts, we found 9 proteins with a significant decrease and 12 proteins with a significant increase. We also found an increase in S-glutathionylation of endothelial nitric oxide synthase in failing female versus male hearts, suggesting an increase in uncoupled nitric oxide synthase in female hearts.
CONCLUSIONS: These findings highlight the importance of nitroso-redox signaling in both physiological and pathological conditions, suggesting a potential target to treat heart failure.