Objective   To analyze the components of proteins released from early ischemic myocardial injury of isolated human heart during hypothermia perfusion and study their physiological processes and fun-ctions.    Methods    Perfusate at 4℃ was harvested from isolated human hearts instantly

30 and 60 min after arrest. Peptide fragments were obtained by removing abundant proteins from samples with MARS columns

desalinizing with a 10 kD molecular sieve

and digesting with enzyme in solution

desalinized again with mRP columns

identified by HPLC-Chip-MS/MS and analyzed. Spectra were established for all proteins released from early ischemic myocardial injury of isolated human heart during hypothermia perfusion. The functions and locations of these proteins were analyzed by GO and EBI

and compared with those of BLAST.    Results    A total of 176 high confidence proteins (score≥8

SPI≥70%) were obtained using the Agilent Spectrum Mill Home software. GO annotations showed that these proteins mainly participated in regulation of the physiological processes of cells

biology and metabolism

with blinding functions

catalytic activities and enzyme-modulating activities. The 106 kD agnoprotein was found to be an exocytosis protein by comparing 106 kD and 29 kD unknown proteins in BLAST and EBI database.    Conclusion   The 176 high-confidence proteins are of great significance for further study of biological markers for early myocardial ischemic injury

and the 106 and 29 kD agnoproteins may be two unknown proteins of early myocardial ischemic markers.