Neuroprotective effects of anthocyanins on apoptosis induced by mitochondrial oxidative stress

Objectives: Mitochondrial oxidative stress (MOS) is a major factor in the underlying pathology of many neurodegenerative diseases. Here, we investigated the neuroprotective effects of a unique class of nutraceutical antioxidants, anthocyanins, against MOS-induced death of cultured cerebellar granule neurons (CGNs). Callistephin and kuromanin are anthocyanins derived from strawberries and black rice, respectively, whose neuroprotective properties have yet to be examined in detail. Methods: Glutathione (GSH)-sensitive MOS and intrinsic apoptosis were induced in CGNs by incubation with the Bcl-2 inhibitor, HA14-1. The effects of anthocyanin co-incubation on CGN survival were assessed. Results: The anthocyanins demonstrated significant protection from MOS-induced apoptosis which was equivalent to that provided by the green tea polyphenol, epigallocatechin 3-gallate; however, neither anthocyanin was as effective as GSH at rescuing CGNs. Inhibition of Bcl-2 caused a significant reduction of mitochondrial GSH which was prevented by the anthocyanins. Furthermore, the anthocyanins inhibited iron-induced lipid peroxidation in rat brain homogenates and prevented cardiolipin oxidation induced by MOS in CGNs. MOS-induced mitochondrial fragmentation and proteolytic cleavage of the optic atrophy 1 (OPA1) fusion GTPase were also attenuated by the anthocyanins. Finally, the anthocyanins significantly enhanced GSH peroxidase activity in a cell-free assay. Discussion: These data show that anthocyanins suppress MOS-induced apoptosis by preserving mitochondrial GSH and inhibiting cardiolipin oxidation and mitochondrial fragmentation. These nutraceutical antioxidants warrant further study as potential therapeutic agents for neurodegenerative diseases caused by MOS.