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Hydrogen sulfide (H2S) generated from H2S-synthesizing enzymes induced nitric oxide (NO) release via nNOS as a defense mechanism during cerebral ischemic injury. Although most studies show that the release of NO during ischemia results in the production of cytotoxic reactive oxygen species (ROS), the roles of NO in ROS generation during ischemia remain unclear. In the present study, the roles of NO were investigated using hippocampal slices as an in vitro ischemia model. Rat hippocampal slices were cut and stored in a medium containing 2 mM Ca2+ for 6 h in a sealed chamber to minimize hypoxia-ischemia. H2S-induced NO release was measured using the fluorescent probe DAF-FM and was activated by the addition of GYY4137 (300 microM). NO alone increased neuronal death compared with control. The addition of 300 microM GYY4137 produced a stronger neuronal death than NO alone, suggesting that NO generation is suppressed during ischemia-reperfusion injury. NO alone increased the production of ROS and the release of lactate dehydrogenase (LDH) during ischemia, and both the increases in ROS and LDH release were inhibited by 300 microM GYY4137. These findings suggest that NO generated during ischemia has cytotoxic effects via the stimulation of ROS production. In contrast, H2S-induced NO release suppressed NO production during ischemia
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