The Study Of Berberine Protects Cardiomyocytes Against Anoxia/Reoxygenation Injury Mediated By AMPK?

Objective:Berberine(BBR),an isoquinoline alkaloid,extracts from the Chinese herb Rhizoma Coptidis which has been reported can attenuate myocardial ischemia/repefusion injury.The major aim is to study the mechanism of BBR against anoxia/reoxygenation(A/R)injury and the inhibitor Compound C used for further clarify whether the protective effect of BBR against A/R injury mediated by AMPK? protein.Method: In this study,rat H9c2 cardiomyocytes were used as experimental vectors,and I/R model in vivo were replicated by cardiomyocytes A/R model in vitro.The experimental was randomly divided into 4 groups,each group repeated 6 times:(1)Control group;(2)A/R group;(3)20?M BBR+A/R group;(4)20?M BBR+1?M Compound C+A/R group.The cell viability was tested by MTS assay.The lactate dehydrogenase(LDH)was detected by Enzyme-linked immunosorbent assay.The AMPK? protein expression was assayed by western blotting analysis.The level of intracellular ROS was tested by DCFH-DA.The result of cardiomyocyte apoptosis was detected by TUNEL assay and Annexin ?-FITC/PI double staining.Results: 1.Compared to A/R group,BBR pretreatment rat H9c2 cardiomyocytes decreased LDH release and activity(P<0.01)and increased the cell viability(P<0.01).2.The result of western blotting showed that 20?M BBR pretreatment rat H9c2 cells up-regulated the expression of AMPK? protein,the difference has statistical significance(P<0.01).3.The result of FCM detection showed that 20?M BBR pretreatment rat H9c2 cardiomyocytes reduced intracellular ROS level(P<0.05).4.The result of Annexin?-FITC/PI double staining showed that 20?M BBR pretreatment H9c2 cells decreased significantly compared to A/R group,and the result of TUNEL was the same of double staining result.5.Compound C abolished the listed protective effect of BBR suffered from A/R injury.Conclusion: Berberine protects H9c2 against A/R injury partly via up-regulating the expression of AMPK??...