Anthocyanins from Hibiscus syriacus L. Inhibit Oxidative Stress-mediated Apoptosis by Activating the Nrf2/HO-1 Signaling Pathway

Hibiscus syriacus L. is widely distributed throughout Eastern and Southern Asia and its root bark has been used as a traditional remedy. Recently, the extracts of H. syriacus L. exerts anti-cancerous, anti-microbial, and anti-inflammatory activities. However, the effect of anthocyanin-rich fraction of H. syriacus L. petals (PS) has not been studied under excessive oxidative stress. In this study, we evaluated the cellular protective effect of PS in HaCaT human skin keratinocytes under hydrogen peroxide ($H_2O_2$)-induced oxidative stress conditions. PS at below $400{\mu}g/ml$ did not show any cell death; however, over $800{\mu}g/ml$ of PS gradually increased cell death. PS at below $400{\mu}g/ml$ significantly inhibited $H_2O_2$-induced apoptosis in HaCaT cells concomitant with downregulation of Bax and upregulation of pro-PARP and p-Bcl-2. Additionally, PS remarkably reversed $H_2O_2$-induced excessive reactive oxygen species (ROS) production and apoptosis, and also significantly inhibited mitochondrial ROS production concomitant with suppression of $H_2O_2$-induced mitochondrial depolarization. $H_2O_2$-mediated ratio of Bax to Bcl-2, and caspase-3 activation were markedly abolished in the presence of PS. Moreover, the inhibition of HO-1 function using zinc protoporphyrin, an HO-1 inhibitor, significantly attenuated the cellular protective effects of PS against $H_2O_2$, indicating the significance of HO-1 in PS mediated cytoprotective effect, which was mediated by activating nuclear factor erythroid 2-related factor-2 (Nrf2). Taken together, our results suggest that cytoprotective effect of PS in HaCaT keratinocytes against oxidative stress-induced apoptosis is mediated by inhibiting cellular and mitochondrial ROS production, which is downregulated by activating Nrf2/HO-1 axis.