锌螯合剂对匹罗卡品致痫小鼠行为学及海马神经元的影响

目的:研究锌螯合剂氯碘羟喹对匹罗卡品致痫小鼠行为学及海马神经元的影响。方法:将CD1小鼠随机分成3组,对照组小鼠腹腔注射等量的生理盐水;癫痫组小鼠腹腔注射匹罗卡品300 mg/kg;氯碘羟喹治疗组(治疗组)小鼠腹腔注射匹罗卡品300 mg/kg,30 min之后腹腔注射氯碘羟喹15 mg/kg。观察各组小鼠的癫痫发作次数。Morris水迷宫实验测定逃避潜伏期和目标象限停滞时间。应用尼氏染色计数海马神经元数量;利用免疫组织化学方法、免疫印迹结合图像分析技术检测凋亡基因caspase-3在各组小鼠海马的表达变化。结果:与对照组相比,其他2组小鼠的癫痫发作次数明显增多,发作级别明显增高;其中,治疗组小鼠的发作次数和发作级别均低于癫痫组。水迷宫实验结果显示,与对照组相比,其他2组小鼠的逃避潜伏期明显延长,目标象限停滞时间明显缩短;其中,治疗组的逃避潜伏期比癫痫组短,目标象限停滞时间比癫痫组长。其他2组小鼠海马神经元数量明显少于对照组;治疗组的海马神经元数量比癫痫组多。其他2组小鼠海马caspase-3表达明显高于对照组,阳性细胞数量增多,光密度增加;治疗组海马caspase-3的表达低于癫痫组,阳性细胞数量减少,光密度降低。结论:锌螯合剂氯碘羟喹能抑制癫痫发作,保护海马神经元,提高癫痫小鼠的学习记忆能力。

Effect of zinc chelating agent on the behaviors and hippocampus neurons of pilocarpine-induced epilepsy mice

Objective :To investigate the effect of clioquinol, a zinc chelating agent, on the ethology and hippocampus neurons of pilocarpine-induced epilepsy mice. Methods :CD1 mice were studied in this experiment.They were divided randomly into 3 groups. Mice were given saline in control group, pilocarpine(300 mg/kg) in epilepsy group and clioquinol(15 mg/kg) 30 min after pilocarpine treatment in clioquinol group intraperitoneally.Then the time and level of epileptic seizure in each group were observed. Morris maze experiment was done 24 h after pilocarpine treatment. The escape latency and time in correct quadrant were checked. Hippocampus tissues were obtained on 7^ th day. Nissl staining was selected to calculate the number of hippocampus neurons. Immunochemistry, immunoblotting and image analysis were used to detect the expression of caspase-3 in the hippocampus of each group. Results :Compared with control group, the time and level of epileptic seizure were higher in both epilepsy group and clioquinol group, which in the clioquinol group was lower than that in epilepsy group. Morris maze experiment showed that the escape latency was longer and the time in correct quadrant was shorter in epilepsy group and clioquinol group than those in control group. Mice in clioquinol group showed shorter escape latency and longer time in correct quadrant than those in epilepsy group. Number of hippocampus neurons was less in epilepsy group and clioquinol group than that in control group, while clioquinol group showed more neurons than epilepsy group. Number of caspase-3 immunopositive neurons and the optical density increased in the hippocampus of epilepsy group and clioquinol group. Mice in the clioquinol group showed less caspase-3 immunopositive neurons and lower optical density than that in epilepsy group. Conclusion :Clioquinol can inhibit epileptic seizure, protect hippocampus neurons and enhance the learning and memory capability of epilepsy mice.