RESUMEN
OBJECTIVE: To investigate the changes in protein of myocardium after hydrogen sulfide delayed preconditioning by using proteomics technology. METHODS: Sixteen Sprague-Dawley rats were randomly assigned to control (group S) or hydrogen sulfide group (group H), n = 8 for each group. Myocardial ischemia/reperfusion injury model (ischemia 30 minutes followed by reperfusion 120 minutes) was reproduced at 24 hours after preconditioning either with normal saline or hydrogen sulfide for proteomics analysis in group S or group H, and the myocardial tissue was harvested. The total proteins were extracted and separated by two dimensional gel electrophoresis (2-DE), and the differential protein expression spots were analyzed with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). RESULTS: Analysis of 2-DE showed that 929 ± 14 protein spots were found in group S and 906 ± 10 protein spots in group H, and the expression of 15 protein spots was different between two groups. These protein spots were chosen to undergo MALDI-TOF-MS analysis, and 11 proteins were preliminarily identified, including DNA ligase, cystathionine gamma-lyase, transcription initiation factor, NADH dehydrogenase, guanine nucleotide-releasing factor, fructose-bisphosphate aldolase A, glycogen synthase kinase-3, electron transfer flavoprotein subunit beta, glutathione S-transferase, soluble calcium-activated nucleotidase and S-adenosylmethionine synthetase. CONCLUSIONS: Hydrogen sulfide delayed preconditioning of myocardium resulted in the changes in protein expression profiles in the myocardium. The differential proteins might function as anti-oxidants, to improve the energy metabolism of myocardium, confer cytoprotection and protection of respiratory chain, thus conferring cardioprotection.
Asunto(s)
Sulfuro de Hidrógeno/farmacología , Precondicionamiento Isquémico Miocárdico , Miocardio/metabolismo , Proteómica/métodos , Animales , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Delayed myocardial preconditioning by volatile anesthetics involves changes in DNA transcription and translation. Mitochondria play a central role in myocardial ischemia/reperfusion (I/R) injury and in ischemic or pharmacologic preconditioning. In this study, we investigated whether there are alterations in myocardial mitochondrial protein expression after volatile anesthetic preconditioning (APC) to examine the underlying mechanisms of delayed cardioprotection. METHODS: Thirty-six Sprague-Dawley rats were randomly assigned to 1 of 3 groups (n = 12 for each group). Rats in the delayed APC group were exposed to sevoflurane (2.5% for 60 minutes) 24 hours before myocardial ischemia was induced. Myocardial ischemia in the I/R and APC groups was induced by left coronary artery occlusion for 30 minutes, followed by 120 minutes of reperfusion. The control group received no treatment. The mitochondria fractions were prepared by differential centrifugation with density gradient isolation for proteomic analysis. Two-dimensional gel electrophoresis and matrix-assisted laser desorption/ionization with time-of-flight mass spectrometry was used to identify differences in the protein expression from mitochondria of the rat hearts. RESULTS: Fifteen differentially expressed mitochondrial proteins between the APC group and I/R group were identified and the expression patterns of 2 of the proteins were confirmed by Western blot analysis. These proteins were associated with mitochondrial substrate metabolism, respiration, and adenosine triphosphate (ATP)/adenosine diphosphate transport. The modifications of the mitochondrial proteome suggest an enhanced capacity of mitochondria to maintain myocardial ATP levels after I/R injury. CONCLUSION: Delayed sevoflurane myocardial preconditioning induces mitochondrial proteome remodeling, which mainly involves proteins that are related to ATP generation and transport. Therefore, proteomic changes related to bioenergetic balance may be the mechanistic basis of delayed anesthetic myocardial preconditioning.