Mechanisms underlying the protective effects of myricetin and quercetin following oxygen-glucose deprivation-induced cell swelling and the reduction in glutamate uptake in glial cells

The protective effects of the flavonoid polyphenols, myricetin and quercetin, were investigated on key features of ischemic injury in cultures including cell swelling and the reduction in glutamate uptake. C6 glial cells were exposed to oxygen-glucose deprivation (OGD) for 5 h and cell swelling was determined 90 min after the end of OGD. OGD-induced swelling was significantly blocked by both quercetin and myricetin although higher concentrations were required for quercetin. OGD-induced free radical production, a contributing factor in cell swelling, was significantly reduced by both myricetin and quercetin. However, depolarization of the inner mitochondrial membrane potential (ΔΨ(m)), the blockade of which generally reduces swelling, was significantly diminished by myricetin, but not quercetin. This indicated that quercetin could reduce swelling despite its inability to prevent depolarization of ΔΨ(m) possibly through other signaling pathways. Increased intracellular calcium ([Ca²+](i)) is an important characteristic of ischemic injury and is implicated in swelling. Both myricetin and quercetin attenuated the increase in [Ca²+](i). Further, a reduction in [Ca²+](i), through the use of nifedipine, nimodipine, verapamil, dantrolene, or BAPTA-AM, significantly reduced OGD-induced cell swelling indicating that one possible mechanism by which such flavonoids attenuate cell swelling may be through regulating [Ca²+](i). OGD-induced decrease in glutamate uptake was attenuated by myricetin, but not quercetin. Cyclosporin A, a blocker of the mitochondrial permeability transition (mPT) pore, but not FK506 (that does not block the mPT), attenuated the decline in glutamate uptake after OGD, indicating the involvement of the mPT in glutamate uptake. Our results indicated that while blockade of ΔΨ(m) may be sufficient to reduce swelling, it may not be a necessary factor, and that flavonoids reduce cell swelling by regulating [Ca²+](i). The differential effects of myricetin and quercetin on OGD-induced reduction on glutamate uptake may be due to their differential effects on mitochondria.