Neuroprotective effects of nicotinamide and 1-methylnicotinamide in acute excitotoxicity in vitro.
Nicotinamide (NAM), an important cofactor in many metabolic pathways, exhibits at high doses neuroprotective abilities of an unclear mechanism. In the present study we evaluated the unknown protective capability of its immediate metabolite 1-methylnicotinamide (MNA) in comparison to NAM in primary cultures of rat cerebellar granule cells (CGC) submitted to acute excitotoxicity. Neurotoxicity was evaluated with propidium iodide staining 24 h after 30 min exposure to glutamate (GLU) and NMDA. NAM and MNA reduced NMDA toxicity only at 25 mM concentration, while neurotoxicity of 0.5 mM GLU was slightly diminished only by 25 mM NAM. Both compounds at 25 mM reduced GLU-induced 45Ca uptake and dose-dependently inhibited NMDA-induced 45Ca accumulation. Neither NAM nor MNA interfered with GLU-evoked intracellular calcium transients evaluated with calcium orange fluorescent probe or inhibited [3H]MK-801 binding to rat cortical membranes. NAM and MNA failed to change GLU-evoked decrease in mitochondrial membrane potential monitored using the fluorescent dye rhodamine 123. Analysis with a hydroperoxide-sensitive fluorescent probe demonstrated significant reduction by 20 and 25 mM MNA, but not NAM, of oxidative stress in cultures after 1 h treatment with GLU. CGC accumulated radiolabelled NAM and MNA in a time and concentration dependent manner, NAM being transported more rapidly. These findings demonstrate that weak neuroprotective ability of MNA in excitotoxicity, accompanied by incomplete stabilization of calcium imbalance and lessening of oxidative stress, is not connected with direct inhibition of NMDA receptors. The exact mechanisms of these effects require further investigation.
Slomka
Marta
M
Department of Neurochemistry, Medical Research Centre, Polish Academy of Sciences, Pawinskiego 5, 02-106 Warsaw, Poland.
Zieminska
Elzbieta
E
Salinska
Elzbieta
E
Lazarewicz
Jerzy W
JW
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D018690:Excitatory Amino Acid Agonists / Q000633:toxicity; D018698:Glutamic Acid / Q000633:toxicity; D066298:In Vitro Techniques; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D016202:N-Methylaspartate / Q000633:toxicity; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism* / Q000473:pathology; D018696:Neuroprotective Agents / Q000494:pharmacology*; D009536:Niacinamide / Q000031:analogs & derivatives* / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats
D018690:Excitatory Amino Acid Agonists; D018696:Neuroprotective Agents; D009536:Niacinamide; D018698:Glutamic Acid; D016202:N-Methylaspartate; D002118:Calcium; C024058:N(1)-methylnicotinamide