The Mechanism Of Puerarin Against Cardiomyocytes Anoxia/Reoxygenation Injury

Objective：Puerarin, a flavonoid glycosidesis, extracted from bean plant kudzu,had been proved to protect cardiomyocytes from cardiomyocytes anoxia/reoxyg-enation injury. PKCε is one of the important endogenous myocardial protectivefactors, is involved in intracellularphysiopathology functions. In our study, we haveestablished the cultured neonatal rat primary cardiomyocytes model with the acutemyocardial anoxia/reoxygenation (A/R) injury model to explore the relationshipbetween the protective effect of puerarin against myocardial anoxia/reoxygenationinjury and the PKCε signal pathway.Methods:The cultured primary cardiomyocytes were divided into five groupsrandomly, each group repeated six times:(1)Control group;(2) A/R group;(3)Pue+A/R group;(4) Pue+εV1-2+A/R group.(5) εV1-2+A/R group.The westernblotting analysis assayed the expression of PKCε and the cell viability was detectedby MTT, The results of MDA content，the activities of CK, LDH, SOD and GSH-Pxwere analyzed by UV spectrophotometer. The standard of intracellular ROS, the mmpand the apoptosis were detected by flow cytometry.Results: The cell viability was increased after preprocessing320μM pueraringroup, the expression of PKCε protein after preprocessing320μM puerarin wassignificantly raised compared to the control group and A/R group（p<0.01); Pue+A/Rgroup, the contents of LDH and CK was significantly decreased, the activities ofSOD and GSH-Px were increased, the contents of MDA was decreased, the contentof ROS was decreased, the MMP were significantly increased, and the apoptosis wasdecreased, compared with A/R group, and all indicators were significant differences(p<0.01); All indicators were significant differences between Pue+A/R group andPue+εV1-2+A/R group (p<0.01), but no significant differences between A/R groupand εV1-2+A/R group（p>0.05).Conclusion: Puerarin against myocardial ischemia-reperfusion injury depend onPKC epsilon signaling pathways, at least in part, depends on degree of PKC epsilon expression level increases.