The Mechanism Of Puerarin Protect Isolated Rat Hearts Against Anoxia/reoxygenation Injury

Objective:The puerarin has been widely used to treat patients with coronary artery diseases,and it has show the protectiveness on hearts’functions.It has profound significance to do the further research of the puerarin on the protect mechanism.We adopted anoxia-reoxygenation injury model in our experiments,pretreated rats with drugs through lumbar injection before isolated hearts perfusion were performed,then investigated the role of puerarin in anoxia-reoxygenation injury,and its’effects on protein expression of PKCε/14-3-3η.Methods:We used Langendorff perfusion system to build the anoxia-reoxygenation injury model.There were 30 rats,and were randomly divided into five groups:① Control group;②/R group;③APC group;④Pue+A/R group;⑤Pue+εV1-2+A/R group,the εV1-2 is a specific inhibitor of PKCe.The signals of the perfusion hearts were collected by the Powerlab system to analyse the hearts functions like LVP,LVEDP,±dp/dt max and CF. The release of CK and LDH in the perfusion solution during A/R were measured by spectrophotometer.Tetrazolium chloride staining revealed the infarct size and the area at risk after reoxygen.Use western blotting to detected the protein expression of PKCε/14-3-3η in each group after reoxygen.Results:①Compared with Control group,A/R group caused great increases of LVP,LVEDP and infarct size(p<0.01),mean while,the releases of CK and LDH were greatly increased at 20min after reoxygen(p<0.01),but great decreases of ±dp/dt max and CF(p<0.01),the protein expression of PKCe and 14-3-3η were also decreased(p<0.01).②In contrast to A/R group,changes of Pue+A/R group on LVP,LVEDP and infarct size were obviously lower(p<0.01),the releases of CK and LDH was evidently decreased at 20min after reoxygen(p<0.01), the ±dp/dt max and CF were increased(p<0.01),the protein expression of PKCε and 14-3-3η were increased(p<0.01). ③Pue+A/R group show the same effect with APC group when compared to A/R group(p<0.01,vs A/R group).④Distinct from Pue+A/R group, the LVP,LVEDP and infarct size of Pue+εV1-2+A/R group were obviously increased(p<0.01),the releases of CK and LDH at 20min after reoxygen were also increased(p<0.01),but the ±dp/dt max and CF were decreased(p<0.01),and the protein expression of PKCε and 14-3-3η were sharply decreased(p<0.01).Conclusion:1. The puerarin preconditioning is effictive in protecting isolated perfusion rat heart against anoxia-reoxygenation injury.2. The cardioprotection of puerarin preconditioning may conducted by up-regulation the protein express of PKCε/14-3-3η.