Protective Effect Of Allicin On Myocardial Ischemia-reperfusion Injury And Its Mechanism

ObjectiveTo explore whether allicin has protective effect on myocardial ischemia-reperfusion injury by inhibiting cardiomyocyte calcium overload and apoptosis,and to further explore whether its regulation of calcium overload is related to PI3K/GRK2/PLC-y/IP3R signal pathway.Materials and methodsIn this study,we investigated the molecular mechanisms by which allicin attenuated the myocardial ischemia-reperfusion injury in vitro and in vivo by focusing on phosphoinositide 3-kinase(PI3K),mitochondrial injury,calcium overload and cardiomyocyte apoptosis.Sixty male Albino Wistar mice were randomly assigned into three groups:myocardial ischemia-reperfusion group,sham-operated group,and myocardial ischemia reperfusion+allicin group.Myocardial ischemia reperfusion and allicin groups were modeled by myocardial ischemia-reperfusion surgery,and mice in the Allicin group were injected by allicin 30 minutes before the reperfusion.Masson staining to dye the heart of mice to evaluate the myocardial infarct area after the reperfusion.Moreover,the TUNEL assay was carried out to examine the anti-apoptotic effect of allicin.The morphology of mitochondria was observed by transmission electron microscope.Differentially expressed genes and pathways of MIRI were analyzed by miRNA microarray analysis and bioinformatics analysis.Real-time quantitative polymerase chain reaction(q-PCR)was performed to validate the selected PI3K gene expression.Immunofluorescence staining and western blot analysis were carried out to detect the effect of allicin treatment on the PI3K in MI/R.A pan-PLC activator,m-3M3FBS,was applied to investigate whether or not the protective effect of allicin on cardiomyocyte apoptosis during MI/R was via the GRK2/PLC-y/IP3R signaling pathway using Fura-2AM staining.ResultsMasson staining and the TUNEL assay revealed that allicin treatment reduced infarct size after M/IR and played the anti-apoptotic role in M/IR.The hierarchical clustering analysis exhibited an intense distinction on differentially expressed miRNAs in Allicin group,as demonstrated in the heat.Gene ontology and Genome analyses indicated the calcium signaling pathway and differentially expressed gene PI3KCA were selected in this study.Immunofluorescence staining and Western Blot analysis showed that allicin increased the phosphorylation level of PI3K and Akt and decreased the expression of p-GRK2,p-CaMKII,PLC-? and p-IP3R during myocardial ischemia-reperfusion injury.Fura-2AM staining showed that allicin could reduce Ca2+overload in cardiomyocytes,and the effect of allicin could be reversed by using pan-PLC activator m-3M3FBS.ConclusionsIt is reported that allicin has a protective effect on myocardial ischemia-reperfusion injury.This effect might be associated with the inhibition of Ca2+overload-induced apoptosis and the inhibition of the PI3K mediated GRK2/PLC-y/IP3R signaling pathway.