The radical scavenger CR‐6 protects SH‐SY5Y neuroblastoma cells from oxidative stress‐induced apoptosis: effect on survival pathways

Abstract

Reactive oxygen species (ROS) and oxidative stress have long been linked to cell death of neurons in many neurodegenerative conditions. However, the exact molecular mechanisms triggered by oxidative stress in neurodegeneration are at present unclear. In the current work we have used the human neuroblastoma SH-SY5Y cell line as a model for studying the molecular events occurring after inducing apoptosis with H 2 O 2 . We show that treatment of SH-SY5Y cells with H 2 O 2 up-regulates survival pathways during early stages of apoptosis. Subsequently, the decline of anti-apoptotic protein levels leads to the activation of the calcium-dependent proteases calpains and the cysteine proteases caspases. Additionally, we demonstrate that CR-6 (3,4-dihydro-6-hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran) acts as a scavenger of ROS and prevents apoptosis by enhancing and prolonging up-regulation of survival pathways. Furthermore, we show that pre-treatment of SH-SY5Y cells with a cocktail containing CR-6, the pan-caspase inhibitor zVAD-fmk (zVal-Ala-Asp-fluoro-methylketone) and the calpain inhibitor SJA6017 confers almost total protection against apoptosis. In summary, the present work characterizes the molecular mechanisms involved in oxidative stress-induced apoptosis in SH-SY5Y cells. Our findings highlight the relevance of CR-6, alone or in combination with other drugs, as potential therapeutic strategy for the treatment of neurodegenerative diseases.

Participation of Oxidative Stress on Rat Middle Cerebral Artery Changes Induced by Focal Cerebral Ischemia: Beneficial Effects of 3,4-Dihydro-6-hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran (CR-6)

“…, NO, and ONOO Ϫ (Montoliu et al, 1999;Sanvicens et al, 2006). Our results show that MCA from ischemic rats presents an enhanced formation of O 2 .…”

“…We also tested the beneficial effects of CR-6, a structurally simple derivative of vitamin E (Casas et al, 1992), with capacity to scavenge oxygen and nitrogen reactive species. CR-6 has previously shown some beneficial effects against oxidative stress in vivo (Miranda et al, 2007) and in vitro (Montoliu et al, 1999;Sanvicens et al, 2006).…”

Redox survival signalling in retina-derived 661W cells

“…This increase in H 2 O 2 levels was completely abrogated by the radical scavenger CR-6 (3,4-dihydro-6-hydroxy-7-methoxy-2,2-dimethyl-1(2H)-benzopyran) 14,17 ( Figure 1b). Furthermore, H 2 O 2 production was significantly reduced by the Nox inhibitor diphenyleneiodonium (DPI), suggesting a role for the Nox system in H 2 O 2 generation.…”

Antioxidant and Anticancer Activities of Methanol Extract of Seeds of Datura stramonium l

“…Oxidative stress is now recognized to be associated with more than 100 diseases, as well as with the normal aging process, especially people diagnosed with Alzheimer's disease (AD). 21 Antioxidants are intimately involved in the prevention of cellular damage the common pathway for cancer, aging, and a variety of diseases.…”