Post-treatment with voltage-gated Na(+) channel blocker attenuates kainic acid-induced apoptosis in rat primary hippocampal neurons.
Injection of rats with kainic acid (KA), a non-N-methyl-D-aspartate (NMDA) type glutamate receptor agonist, induces recurrent (delayed) convulsive seizures and subsequently hippocampal neurodegeneration, which is reminiscent of human epilepsy. The protective effect of anti-epileptic drugs on seizure-induced neuronal injury is well known; however, molecular basis of this protective effect has not yet been elucidated. In this study, we investigated the effect and signaling mediators of voltage-gated Na(+) channel blockers (Lamotrigine, Rufinamide, Oxcarbazepine, Valproic Acid, and Zonisamide) on KA-induced apoptosis in rat primary hippocampal neurons. Exposure of hippocampal neurons to 10 μM KA for 24 h caused significant increases in morphological and biochemical features of apoptosis, as determined by Wright staining and ApopTag assay, respectively. Analyses showed increases in expression and activity of cysteine proteases, production of reactive oxygen species (ROS), intracellular free [Ca(2+)], and Bax:Bcl-2 ratio during apoptosis. Cells exposed to KA for 15 min were then treated with Lamotrigine, Rufinamide, Oxcarbazepine, Valproic Acid, or Zonisamide. Post-treatment with one of these anti-epileptic drugs (500 nM) attenuated production of ROS and prevented apoptosis in hippocampal neurons. Lamotrigine, Rufinamide, and Oxcarbazepine appeared to be less protective when compared with Valproic Acid or Zonisamide. This difference may be due to blockade of T-type Ca(2+) channels also by Valproic Acid and Zonisamide. Our findings thus suggest that the anti-epileptic drugs that block both Na(+) channels and Ca(2+) channels are significantly more effective than agents that block only Na(+) channels for attenuating seizure-induced hippocampal neurodegeneration.
Department of Neurosciences, Medical University of South Carolina, 96 Jonathan Lucus Street, Charleston, SC 29425, USA.
Arabinda
A
Das
Misty
M
McDowell
Casey M
CM
O'Dell
Megan E
ME
Busch
Joshua A
JA
Smith
Swapan K
SK
Ray
Naren L
NL
Banik
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002154:Calpain / Q000378:metabolism; D002478:Cells, Cultured; D003124:Colorimetry; D004789:Enzyme Activation; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015640:Ion Channel Gating; D007608:Kainic Acid / Q000037:antagonists & inhibitors* / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D051381:Rats; D017382:Reactive Oxygen Species; D026941:Sodium Channel Blockers / Q000494:pharmacology*; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D017382:Reactive Oxygen Species; D026941:Sodium Channel Blockers; D051028:bcl-2-Associated X Protein; D002154:Calpain; D007608:Kainic Acid