Resveratrol Protects Cardiomyocytes Against Anoxia/Reoxygenation Injury By Promoting VDAC1 De-phosphorylation Modification

Objective: As we previously demonstrated,VDAC1,a protein located in the mitochondrial outer membrane,is involved in the effects of resveratrol on mitochondrial permeability transition pore(mPTP).However,the regulation mechanism remains obscure.In this study,we aim to explore the effect of resveratrol on VDAC1 phosphorylated modification and the relationship with Akt-GSK3? pathway in H9c2 cardiomyocytes subjected to anoxia/reoxygenation(A/R)injury.Methods: In this study,an in vitro I/R model was replicated on H9c2 cardiomyocytes by A/R treatment.p-Akt(Ser473),Akt,p-GSK3?(Ser9),GSK3?,VDAC1 protein expression were detected by Western Blotting;p-VDAC1,HK2-VDAC1 complexes,and Bax-VDAC1 complexes expression were detected by Co-Immunoprecipitation.According to the kit,the activity of CPK,LDH were measured by UV spectrophotometer.The levels of intracellular ROS,mitochondrial membrane potential(??m)and apoptosis were detected by flow cytometry.Results: 1.After A/R injury,the expression of VDAC1,p-VDAC1 and Bax-VDAC1 complexes expression were increased compared with control group,the expression of p-Akt(Ser473),p-GSK3?(Ser9)and HK2-VDAC1 complexes were remarkably dropped compared to control group(p<0.01).2.Preconditioning with Resveratrol,the expression of VDAC1,p-VDAC1 and Bax-VDAC1 complexes expression were dropped compared with A/R grou,the expression of p-Akt(Ser473),p-GSK3?(Ser9)and HK2-VDAC1 complexes were remarkably increased compared to A/R group(p<0.01).3.To compared with Resveratrol pretreatment group,Akt-GSK3? pathway was inhibited when used the specific inhibitor of Akt inhibitor IV,come with VDAC1 phosphorylation level was rised,the cell viability was remarkably decreased,the activity of CPK and LDH were ascended.While VDAC1 de-phosphorylated modification were inhibited by Akt inhibitor IV,A/R injury would induced ROS generation,decreased the stability of ??m,increased the release of cytochrome c from the mitochondria into cytosol and finally induced cell apoptosis(p<0.01).Conclusion: Resveratrol activates Akt-GSK3 ? pathway,prompts VDAC1 de-phosphorylated modification,subsequently inhibit the mitochondrial apoptosis pathway,reduce cell apoptosis,finally protects cardiomyocytes against A/R injury.