Abstract
BackgroundTraditional studies of the cardiac proteome have mainly investigated in an animal model by two-dimensional gel electrophoresis (2-DE). However, the results have not been of satisfactory quality for an understanding of the underlying mechanism. Recent quantitative proteomic methods have been improved to overcome these limitations. To comprehensively study the cardiac proteome in a rat model of ischemia-reperfusion (IR), we developed a tandem mass tag (TMT)-based quantitative proteomic strategy. Furthermore, using this strategy, we examined the molecular mechanisms underlying the prevention of myocardial infarction by the intake of Triticum aestivum L. extract (TALE), a representative dietary fiber grain.MethodsCardiac proteomes were analyzed by 2-DE as a gel-based approach, and TMT labeling coupled with two-dimensional liquid chromatography (2D-LC) and tandem mass spectrometry (MS/MS) as a non-gel-based quantitative approach. Additionally, gene ontology annotation was conducted by PANTHER database. Several proteins of interest were verified by a Western blot analysis.ResultsTotal 641 proteins were identified commonly from two independent MS datasets using 2D-LC MS/MS. Among these, we identified 151 IR-related proteins that were differentially expressed between the sham-operation group and IR group, comprising 62 up-regulated proteins and 89 down-regulated proteins. Most of the reduced proteins were involved in metabolic processes. In addition, 57 of the IR-related proteins were affected by TALE intake, representing 25 up-regulated proteins and 32 down-regulated proteins. In particular, TALE intake leads to a switch in metabolism to reduce the loss of high-energy phosphates and the accumulation of harmful catabolites (especially reactive oxygen species (ROS)) and to maintain cytoskeleton balance, leading to a reduction in cardiac IR injury.ConclusionsOur study provides a comprehensive proteome map of IR-related proteins and potential target proteins and identifies mechanisms implicated in the prevention of myocardial infarction by TALE intake in a rat IR model.
Highlights
Traditional studies of the cardiac proteome have mainly investigated in an animal model by twodimensional gel electrophoresis (2-DE)
Our study provides a comprehensive proteome map of IR-related proteins and potential target proteins and identifies mechanisms implicated in the prevention of myocardial infarction by Triticum aestivum L. extract (TALE) intake in a rat IR model
Experimental design for the cardiac proteome analysis To study the physiological change in the animals due to myocardial infarction at the proteome level, a typical Two-dimensional gel electrophoresis (2-DE) approach has been used in many proteomic studies [7,8,9,10]
Summary
Traditional studies of the cardiac proteome have mainly investigated in an animal model by twodimensional gel electrophoresis (2-DE). To comprehensively study the cardiac proteome in a rat model of ischemia-reperfusion (IR), we developed a tandem mass tag (TMT)-based quantitative proteomic strategy. Using this strategy, we examined the molecular mechanisms underlying the prevention of myocardial infarction by the intake of Triticum aestivum L. extract (TALE), a representative dietary fiber grain. Ischemia results from an imbalance between myocardial oxygen demand and supply, causing cardiac dysfunction, arrhythmias, myocardial infarction, and sudden death. Early diagnostic markers of ischemic heart disease and myocardial infarction and pharmacological postischemic treatments have been widely investigated to reduce infarction size and mortality after IR injury [3, 4]. The pathogenesis of myocardial infarction and its progression to heart failure are complex multifactorial processes that largely depend on the size of the infarcted myocardium
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