Objective To explore the molecular mechanism of c-jun antisense gene transfection in protecting cardiomyocytes from injury of hypoxia and burn serum treatment. Methods Burn sera were collected from wistar rat with 30% total body surface area(TBSA) Ⅲ degree burn injury, and the mixture gas containning 1% O 2 was used as hypoxia model. The c-jun antisense gene recombinant vector was constructed. Neonatal wistar rat cardiomyocytes cultured in vitro were treated with hypoxia and burn sera. c-jun antisense gene recombinant vector was transfected into the cultured cardiomyocytes. Expressions of c-jun, PKC? and JNK were detected with western blot in the transfected and non-transfected cardiomyocytes. Results Expression levels of c-jun, PKC? and JNK significantly increased in the non-transfected cardiomyocytes when treated by hypoxia and burn sera, up to maximum 24 hour after the treatment. Expression levels of c-jun, PKC? and JNK in the transfected cardiomyocytes decreased significantly compared with non-transfected cells. Conclusions The transfection of the c-jun antisense gene recombinant vector protected cardiomyocytes from injury of hypoxia and burn sera treatment via down-regulating PKC? and JNK expressions.