Protective effect of grape seed extract against oxidative stress-induced cell death in a staurosporine-differentiated retinal ganglion cell line

Abstract

OBJECTIVE: Grape seed extract (GSE) is a potent antioxidant. We examined the effect of GSE on oxidative stress-induced cell death in a transformed retinal ganglion cell line, RGC-5.

METHODS: Staurosporine-differentiated RGC-5 (ssdRGC-5) cells obtained by treating RGC-5 cells with 1 µM staurosporine were incubated with GSE for 2 h and then exposed to buthionine sulfoximine plus glutamate (B/G) for 24 h. Cell death was detected using the LIVE/DEAD viability assay and the type of cell death was evaluated using fluorescein isothiocyanate-conjugated Annexin-V/propidium iodide staining. To investigate the mechanism underlying cell death, we determined the caspase-3 activity and level of reactive oxygen species (ROS) formation.

RESULTS: Treatment of ssdRGC-5 cells with B/G increased intracellular ROS and induced apoptosis (not necrosis) with increasing caspase-3 activity. GSE rescued the ssdRGC-5 cells from oxidative stress-induced cell death by inhibiting both intracellular ROS production and caspase-3 activation.

CONCLUSION: GSE had a neuroprotective effect against oxidative stress-induced apoptotic death in ssdRGC-5 cells.