ABSTRACT
AIM:To explore the role of aloperine in ischemia-reperfusion(I/R)-induced H9c2 cardiomyocyte injury and inflammation.METHODS: The H9c2 cardiomyocytes were cultured under hypoxia and re-oxygenation condi-tions to simulate ischemia-reperfusion(SI/R)injury.After treatment with aloperine at various doses,the cell viability was measured by MTT assay.The cell apoptosis was analyzed by flow cytometry.Simultaneously,the levels of lactate dehydro-genase(LDH),malonaldehyde(MDA)and caspase-3 activity were detected by the commercial kits.The levels of inflam-matory cytokines were also detected by ELISA.Moreover,the effects of aloperine on the activation of PI 3K/AKT signaling pathway were determined by Western blot.RESULTS:Pre-treatment with aloperine remarkably abated the inhibitory effect of SI/R on H9c2 cell viability,and decreased the elevations of LDH and MDA triggered by SI /R(P<0.05).Pre-treat-ment with aloperine dramatically suppressed the cell apoptosis induced by SI /R treatment(P<0.05), concomitant with the decrease in caspase-3 activity and increase in Bcl-2/Bax ratio(P<0.05).In contrast to SI/R group,aloperine treat-ment notably restrained the concentrations of pro-inflammatory cytokines, including interleukin-6, tumor necrosis factor-α and interleukin-1β(P<0.05).Furthermore, aloperine remarkably increased the protein levels of p-PI3K and p-AKT. While blocking the PI3K/AKT pathway with its specific inhibitor LY294002, the viability-promoting, anti-apoptotic and anti-inflammatory effects of aloperine on the H 9c2 cells were obviously attenuated(P<0.05).CONCLUSION: Alope-rine protects against cardiomyocytes from I/R injury and inhibits inflammatory responses by activating the PI 3K/AKT signa-ling pathway,implying a potential benefic role of aloperine against myocardial I /R injury.