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The World Health Organization recommended diagnostic criteria for myocardial ischemia include three aspects, medical history and symptoms, the performance of the ECG (electrocardiogram, ECG) and biochemical markers. Myocardial ischemia clinical performance is not typical of the diversity of the many occult symptoms difficult to detect; ECG recording electrophysiological activity of the heart, can only be recorded in the minutes of myocardial ischemia, low sensitivity, literature reported ECG diagnosis of myocardial ischemia sensitivity of less than 50%. Therefore, a sensitive, reliable, specific biomarkers that can be detected in the early stages of myocardial ischemic injury will help early diagnosis and prompt treatment of acute myocardial ischemia in patients. In recent years, the rapid development of proteomics technology, a new technology platform for the exploration of biomarkers for early diagnosis of myocardial ischemia. Proteomics by a comparative analysis of the proteome in normal individuals and pathological individuals, find some disease-specific protein molecules, thus providing molecular markers for the early diagnosis of the disease. Purpose: cryogenic people isolated perfused cardiac ischemic injury early proteomics research, analysis cryogenic isolated perfused ischemic heart early protein release and physiological processes and functional analysis of proteins collected for final screened can lay the foundation for clinical diagnosis of myocardial ischemic injury early specific biomarkers. Method: collection of isolated cardiac arrest after 30min and 60min after cold perfusion cardioplegia the first MARS column to remove high-abundance proteins in the sample, concentrated 10KD molecular sieve desalination solution digestion, peptide MRP column again desalted, lyophilized, by HPLC-Chip-MS/MS identified by mass spectrometry using Agilent's Spectrum Mill professional statistical analysis processing software IPI protein database search by GO annotation and EBI database of protein function, positioning analysis and BLAST alignment. Results: 1, identification of high confidence protein 176 (Score ≧ 8, SPI ≧ 70%). MB, LDH, CK, CA, H-FABP, GOT, as well as A1BG and Actin (actin) of Ceruloplasmin (ceruloplasmin), and two unknown proteins 106KD protein, 29KD protein. High confidence protein 87 proteins involved in cellular processes, including mitochondria, cytoskeleton, organelle organization; 74 proteins involved in biological regulation, including the regulation of biological and metabolic processes, and regulation of gene expression; 71 protein involved in the metabolic process, including anabolic catabolic processes and biological processes, as well as the protein modification process; 44 involved in the response to stimuli, including the stress response, immune response, etc.; 34 involved in the positioning is established, including the ion transport and protein transit; 31 proteins involved in developmental processes, including cell differentiation and cell development; including biological adhesion process of multi-cellular biological processes, physiology. With binding function in terms of functionality, the 98 proteins, 47 proteins have catalytic activity, 20 protein has enzyme regulator activity, structural molecule activity, transporter activity, transcriptional activity and antioxidant activity in other functions. 3, identified to 106KD protein sequence length 1703AA, unc-13 protein sequence homology to reach 100%; the 29KD protein sequence length 276AA, LOC339524 protein homology of 99%, the local contrast found in 117 position in the sequence space. Conclusion: 1, detected high confidence protein 176, early onset of cardiac ischemia release of the protein with the further understanding of physiological processes and functional analysis, for the final screening to clinical diagnosis of myocardial ischemic injury specific biomarkers of early foundation. 2, high confidence protein the cTn, that cryogenic isolated perfused heart 1h myocardial necrosis, and to ensure that the process of myocardial ischemic injury in early experiments. 3, A1BG, Actin, Ceruloplasmin acute phase proteins in myocardial ischemia 1h, but did not find CRP, TNF-α, IL-6 and other inflammatory factors, closely related to the acute phase proteins C-reactive protein and other inflammatory cytokines, we consider early markers of inflammation in myocardial ischemic injury is secondary to the acute phase after reactive protein, but the exact relationship needs further study. 4, Actin detected in this study, Ceruloplasmin, C6 and ZHAO Hui-hui applications such as high-resolution ion mobility mass spectrometry with nanoliter ultra-high performance liquid chromatography to find Unstable angina QDBS biomarkers consistent. Detected two unknown proteins, 106KD and 29KD protein. 106KD protein sequence homology to belong to the unc-13 protein family, initially speculated 106KD protein is a cellular role of the protein, the 29kD protein is a characteristic protein is currently unclear nature. The experiments on human low temperature hearts from ischemia early release protein research, but is unable to carry out due to the specimen source and research funding large samples; 106KD and 29KD two unknown protein is missing in all myocardial blood injury early were able to express, and myocardial cell structure composed of metabolism and myocardial cell function will be the focus of our next step, and the main contents.
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