Objective:To construct and identify adenovirus vector containing the chromogranin A 1-76 segment (CGA1-76),or vasostatin-1 (VS-1) gene,and to observe its expression in cardiomyocytes of rats, so as to investigate the protective effects of VS-1 transgenic therapy on myocardial hypoxia/reoxygenation (H/R) injury. Methods:(1) The primers of CGA1-76 cDNA was designed and used for PCR amplification. The products were then cloned into adenovirus shuttle plasmid pAdTrack and linearized by enzyme Pme Ⅰ; the resultant plasmid was co-transfected into E.coli BJ5183 cells with adenovirus backbone plasmid pAdEasy-1 for homologous recombination. Then the recombinant plasmid was identified,linearized and packaged with QBI-293A cells to amplify the recombinant adenovirus Ad-VS-1,which was then used to infect H9c2 cardiomyocytes. Western blotting was used to detect the expression of VS-1 protein expression. (2)Myocardial H/R model was established and the cells were divided into 4 groups:blank group,H/R group,mock infection + H/R group,and Ad-VS-1 infection + H/R group. The viability of cardiomyocytes,the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were measured. Results:(1) The recombinant plasmid was constructed successfully as confirmed by PCR,sequencing and enzyme digestion. Western blotting confirmed the protein expression of VS-1 in the H9c2 cells. (2)The viability of cardiomyocytes and the activity of SOD in H/R group were obviously lower than those of the blank group; the level of MDA was higher than that of the blank group. Transfection with VS-1 increased the cardiomyocytes viability and SOD activity in the H/R model group and decreased the production of MDA. Conclusion:Ad-VS-1 has been successfully constructed,and transfection with it can protect rat cardiomyocytes from H/R injury,which is related to the anti-oxidation process.