The role of superoxide dismutase and alpha-tocopherol in the development of seizures and kindling induced by pentylenetetrazol - influence of the radical scavenger alpha-phenyl-N-tert-butyl nitrone.
Previous experiments have shown that the generation of free hydroxyl radicals in rat brain homogenates is increased following pentylenetetrazol (PTZ) kindling. The present study was performed in order to evaluate the involvement of endogeneous radical defence systems as the superoxide dismutase (SOD) and the level of alpha-tocopherol, an important lipid-soluble and membrane-bound antioxidant in brain homogenate of rats after acute seizure and kindling induced by PTZ. The activities of the total SOD were significantly reduced after acute seizure and tend towards an enhancement in kindled animals. Western blot analysis shows an upregulation of Mn-SOD in rat brain homogenates after kindling. The level of the chain-breaking antioxidant alpha-tocopherol was reduced in acutely convulsing rats and was not modified in kindled rats. Second, we studied the influence of exogeneously supplied radical scavenger alpha-phenyl-N-tert-butyl-nitrone (PBN) on seizure and kindling following PTZ treatment. After a single injection of PTZ at a dose evoking clonic-tonic seizures, PBN did not modify either the formation of free hydroxyl radicals measured by the levels of 2,3-dihydroxybenzoic acid (DHBA) and 2,5-DHBA or the susceptibility to PTZ. In the kindling group, subchronic treatment with PBN (over a period of 4 weeks) prevented the increase in the formation of free hydroxyl radicals, and the susceptibility to PTZ was transiently decreased during the development of kindling, but PBN did not influence the susceptibility to PTZ in fully kindled rats. Pretreatment with PBN increased the activities of total SOD and the protein content of Mn-SOD and decreased the level of alpha-tocopherol in comparison to saline controls. The results suggest that the formation of free hydroxyl radicals is not reflected by an enhanced susceptibility to PTZ classified according to the modified RACINE scale. Additionally, it may be assumed that the increased generation of hydroxyl radicals in kindled animals is not primary caused by an exhaustion of both the defence systems measured. Adaptive mechanisms, as the induction of Mn-SOD, may be taken into consideration to counteract oxidative stress-mediated free radical formation.
Rauca
Christine
C
Department of Pharmacology and Toxicology, Faculty of Medicine, Otto-von-Guericke University Magdeburg, Leipziger Street 44, 39120 Magdeburg, Germany.
Wiswedel
Ingrid
I
Zerbe
Renate
R
Keilhoff
Gerburg
G
Krug
Manfred
M
D000704:Analysis of Variance; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003292:Convulsants; D003497:Cyclic N-Oxides; D004195:Disease Models, Animal; D004334:Drug Administration Schedule; D005841:Gentisates / Q000378:metabolism; D062385:Hydroxybenzoates / Q000378:metabolism; D007696:Kindling, Neurologic / Q000187:drug effects*; D008297:Male; D018696:Neuroprotective Agents / Q000627:therapeutic use*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II; D009589:Nitrogen Oxides / Q000627:therapeutic use*; D010433:Pentylenetetrazole; D051381:Rats; D017208:Rats, Wistar; D012640:Seizures / Q000139:chemically induced / Q000188:drug therapy*; D013482:Superoxide Dismutase / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D024502:alpha-Tocopherol / Q000378:metabolism*
D003292:Convulsants; D003497:Cyclic N-Oxides; D005841:Gentisates; D062385:Hydroxybenzoates; D018696:Neuroprotective Agents; D009589:Nitrogen Oxides; C029217:phenyl-N-tert-butylnitrone; C009135:2,3-dihydroxybenzoic acid; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II; C496318:Nos2 protein, rat; D013482:Superoxide Dismutase; D024502:alpha-Tocopherol; C010925:2,5-dihydroxybenzoic acid; D010433:Pentylenetetrazole