Protective effect of nicotinamide on neuronal cells under oxygen and glucose deprivation and hypoxia/reoxygenation.
Nicotinamide (vitamin B(3)) reduces the infarct volume following focal cerebral ischemia in rats; however, its mechanism of action has not been reported. After cerebral ischemia and/or reperfusion, reactive oxygen species (ROS) and reactive nitrogen species may be generated by inflammatory cells through several cellular pathways, which can lead to intracellular calcium influx and cell damage. Therefore, we investigated the mechanisms of action of nicotinamide in neuroprotection under conditions of hypoxia/reoxygenation. Results showed that nicotinamide significantly protected rat primary cortical cells from hypoxia by reducing lactate dehydrogenase release with 1 h of oxygen-glucose deprivation (OGD) stress. ROS production and calcium influx in neuronal cells during OGD were dose-dependently diminished by up to 10 mM nicotinamide (p < 0.01). This effect was further examined with OGD/reoxygenation (H/R). Cells were stained with the fluorescent dye 4,6-diamidino-2-phenylindole (DAPI) or antibodies against anti-microtubule-associated protein-2 and cleaved caspase-3. Apoptotic cells were studied using Western blotting of cytochrome c and cleaved caspase-3. Results showed that vitamin B(3) reduced cell injury, caspase-3 cleavage and nuclear condensation (DAPI staining) in neuronal cells under H/R. In addition, nicotinamide diminished c-fos and zif268 immediate-early gene expressions following OGD. Taken together, these results indicate that the neuroprotective effect of nicotinamide might occur through these mechanisms in this in vitro ischemia/reperfusion model.
Shen
Chiung-Chyi
CC
Department of Neurosurgery, Taichung Veterans General Hospital, Taichung, Taiwan, ROC.
Huang
Hsueh-Meei
HM
Ou
Hsiu-Chung
HC
Chen
Huan-Lian
HL
Chen
Wen-Chi
WC
Jeng
Kee-Ching
KC
D000818:Animals; D017209:Apoptosis; D002545:Brain Ischemia / Q000517:prevention & control*; D002118:Calcium / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology; D005947:Glucose / Q000378:metabolism; D000860:Hypoxia; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008954:Models, Biological; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D009536:Niacinamide / Q000494:pharmacology*; D010100:Oxygen / Q000378:metabolism; D020011:Protective Agents / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism; D015424:Reperfusion
D020011:Protective Agents; D017382:Reactive Oxygen Species; D009536:Niacinamide; D007770:L-Lactate Dehydrogenase; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D005947:Glucose; D010100:Oxygen; D002118:Calcium