Plumbagin protects H9c2 cardiomyocytes against TBHP‑induced cytotoxicity by alleviating ROS‑induced apoptosis and modulating autophagy

Abstract

Plumbagin has been previously reported to alleviate myocardial ischemia/reperfusion injury in vivo. In this study, we analyzed the potential role of plumbagin against hydrogen peroxide-induced injury in cardiomyocytes. In the present study, plumbagin (PLB) was used to evaluate its cytoprotective property in H9c2 cardiomyocytes against tertiary butyl hydrogen peroxide (TBHP, 75 μM) induced ROS-mediated oxidative stress and apoptosis. Our results implicate that pretreatment with PLB (5, 10 or 20 μM) notably restored viabilities in TBHP-induced H9c2 cells (p<0.01). Also PLB treatment significantly decreased creatine kinase (CK) (p<0.01) and lactate dehydrogenase (LDH) activity (p<0.01). TBHP induced apoptosis and oxidative stress in cultured cardiomyocyte, whereas PLB pretreatment significantly reduced TBHP-induced apoptosis rate (p<0.01) and ROS level (p<0.01). Furthermore, PLB resulted in decrease in the expressions of cleaved caspase 3, nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme 4 (NOX4) and phospho-p38 MAPK in TBHP-induced H9c2 cells. And the active marker of autophagosomes, LC3-II/LC3-I was elevated following treatment with PLB. These findings indicated that PLB may induce autophagy. The present study shows the protective role of PLB against TBHP-induced cardiomyocyte injury via alleviation of ROS-mediated apoptosis and induction of autophagy.