Ginkgolide B Maintains Calcium Homeostasis in Hypoxic Hippocampal Neurons by Inhibiting Calcium Influx and Intracellular Calcium Release

Ginkgolide B (GB), a terpene lactone and active ingredient of Ginkgo biloba, shows protective effects in neuronal cells subjected to hypoxia. We investigated whether GB might protect neurons from hypoxic injury through regulation of neuronal Ca(2+) homeostasis. Primary hippocampal neurons subjected to chemical hypoxia (0.7 mM CoCl(2)) in vitro exhibited an increase in cytoplasmic Ca(2+) (measured from the fluorescence of fluo-4), but this effect was significantly diminished by pre-treatment with 0.4 mM GB. Electrophysiological recordings from the brain slices of rats exposed to hypoxia in vivo revealed increases in spontaneous discharge frequency, action potential frequency and calcium current magnitude, and all these effects of hypoxia were suppressed by pre-treatment with 12 mg/kg GB. Western blot analysis demonstrated that hypoxia was associated with enhanced mRNA and protein expressions of Ca(v)1.2 (a voltage-gated Ca(2+) channel), STIM1 (a regulator of store-operated Ca(2+) entry) and RyR2 (isoforms of Ryanodine Receptor which mediates sarcoplasmic reticulum Ca(2+) release), and these actions of hypoxia were suppressed by GB. Taken together, our in vitro and in vivo data suggest that GB might protect neurons from hypoxia, in part, by regulating Ca(2+) influx and intracellular Ca(2+) release to maintain Ca(2+) homeostasis.