艾地苯醌抗癫痫作用的用药时间和剂量研究

Study on optimal time and dose of idebenone for antiepileptic effect

目的研究艾地苯醌抗癫痫作用的最适用药时间与剂量。方法SD大鼠随机分为对照组、模型组(经典氯化锂-匹罗卡品诱导法)、12 h+60 mg/kg组、12 h+120 mg/kg组、24 h+60 mg/kg组、24 h+120 mg/kg组。12 h+60 mg/kg组和12 h+120 mg/kg组癫痫持续状态12 h后分别给予60 mg/kg和120 mg/kg艾地苯醌干预,24 h+60 mg/kg组和24 h+120 mg/kg组癫痫持续状态24 h后分别给予60 mg/kg和120 mg/kg艾地苯醌干预,记录给药后癫痫发作频率、时间。水迷宫实验检测不同时间、不同剂量艾地苯醌对大鼠学习、记忆能力影响。通过Western blot检测海马N-甲基-D-天冬氨酸受体2B(N-methyl-D-aspartate receptor 2B,NMDAR2B)磷酸化水平的变化,检测大鼠棘波频率。结果与模型组相比,12 h+60 mg/kg组大鼠癫痫发作频率、时间、大鼠记忆能力无显著变化(均P>0.05),大鼠海马组织NMDAR2B磷酸化水平和脑电波频率也无显著变化(P>0.05)。与模型组相比,12 h+120 mg/kg组大鼠癫痫发作频率显著降低(P<0.05)、发作时间显著减少(P<0.05),大鼠记忆能力显著改善(P<0.05),大鼠海马组织中NMDAR2B磷酸化水平及棘波频率显著下降(均P<0.05)。与模型组相比,24 h+60 mg/kg组和24 h+120 mg/kg组大鼠癫痫发作频率显著降低(P<0.05)、发作时间显著减少(P<0.05),大鼠记忆能力改善显著(P<0.05)。与12 h+120 mg/kg组相比,24 h+60 mg/kg组和24 h+120 mg/kg组大鼠癫痫发作频率、时间和大鼠记忆能力无显著差异(均P>0.05)。Western blot结果表明,与模型组相比,24 h+60 mg/kg组和24 h+120 mg/kg组大鼠海马组织NMDAR2B磷酸化水平显著下降(P<0.005);与12 h+120 mg/kg组相比,24 h+60 mg/kg组和24 h+120 mg/kg组NMDAR2B磷酸化水平无显著变化(P>0.05)。结论在氯化锂-匹罗卡品诱导的大鼠癫痫模型中,癫痫终止后12 h给予120 mg/kg艾地苯醌治疗可以达到较好的抗癫痫效果,这一作用可能与NMDAR2B受到抑制有关。

Objective To study the optimal time and dose of idebenone for the antiepileptic effect.Methods SD rats were randomly assigned into control group,model group,12 h+60 mg/kg group,12 h+120 mg/kg group,24 h+60 mg/kg group,24 h+120 mg/kg group.The epilepsy model was induced by classical lithium chloride-pilocarpine induction method.Rats in 12 h+60 mg/kg group and 12 h+120 mg/kg idebenone group were given 60 mg/kg and 120 mg/kg idebenone after 12 h epilepsy,respectively.Rats in 24 h+60 mg/kg group and 24 h+120 mg/kg idebenone group were given 60 mg/kg and 120 mg/kg idebenone after 24 h epilepsy,respectively.The frequency and the duration of epileptic seizures were recorded.The changes of learning and memory ability were evaluated by water maze test.The phosphorylation level of NMDAR2B(N-methyl-D-aspartate receptor 2B)in hippocampus was detected by Western blot,and the frequency of the brain waves was detected.Results There was no statistical difference in the seizure frequency and duration,the memory ability,the phosphorylation level of NMDAR2B in hippocampus and the frequency of brain waves between model group and 12 h+60 mg/kg(all P>0.05).Compared with model group,the seizure frequency and the seizure duration of rats were significantly decreased in 12 h+120 mg/kg group(P<0.05),the memory ability of rats was significantly improved(P<0.05),and the phosphorylation level of NMDAR2B and the frequency of spines in hippocampal tissue of rats were significantly decreased(P<0.05).Compared with model group,the seizure frequency and the seizure duration were significantly decreased in 24 h+60 mg/kg group and 24 h+120 mg/kg group(P<0.05),and the memory ability was significantly improved(P<0.05).Compared with 12 h+120 mg/kg group,the seizure frequency,the seizure duration and the memory ability of rats showed no significant change in 24 h+60 mg/kg group and 24 h+120 mg/kg group(all P>0.05).Western blot results showed that compared with model group,the phosphorylation level of NMDAR2B in hippocampus was significantly decreased in 24 h+60 mg/kg group and 24 h+120 mg/kg group(P<0.005).Compared with 12 h+120 mg/kg group,the phosphorylation level of NMDAR2B showed no significant change in 24 h+60 mg/kg group and 24 h+120 mg/kg group(P>0.05).Conclusion The antiepileptic effect is better in the lithium chloride-pilocarpine induced epilepsy rat models after treatment with 120 mg/kg idebenone at 12 h after seizure termination,which may be related to the inhibition of NMDAR2B receptor function.