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目的探讨外源性硫化氢对癫痫(epilepsy,EP)所致大鼠脑认知功能障碍的保护作用及机制。方法利用氯化锂-匹罗卡品腹腔注射复制雄性SD大鼠96只,6周龄,体质量180～220 g。癫痫模型,大鼠分为正常对照组(NC组)、癫痫模型组(EP组)、癫痫+硫氢化钠组(NaHS组);采用Morris水迷宫检测大鼠的空间学习记忆能力;实时荧光定量PCR检测海马中的胱硫醚-β-合酶(CBS)mRNA和CREB mRNA表达变化。结果 EP组的空间辨别学习记忆能力与NC组比较明显减退:Morris水迷宫定位航行实验中逃避潜伏期明显延长(P<0.01),第3象限探索有效时间显著缩短(P<0.05)、第3象限游泳距离占总距离的百分比以及穿越平台次数明显减少(P<0.01);NaHS可提高癫痫大鼠的学习记忆能力,与EP组比较,空间学习记忆能力各项检测指标均有显著差异(P<0.05,P<0.01)。7、30 d时EP组大鼠海马CBSmRNA表达水平明显低于NC组(P<0.05),而NaHS组CBS mRNA表达水平明显高于EP组(P<0.05);EP组海马CREBmRNA表达在7、30 d时均明显低于NC组(P<0.05),而NaHS组CREB mRNA在24 h、7、30 d时表达显著高于EP组(P<0.05),与NC组相比,7、30 d时均无差异(P>0.05)。结论外源性H2S可能通过上调癫痫大鼠其海马CBS、CREBmRNA表达来改善癫痫大鼠的学习记忆能力。
Objective To investigate the protective effect and mechanism of exogenous hydrogen sulfide on cognitive impairment induced by epilepsy (EP) in rats. Methods Ninety-six male Sprague-Dawley rats were injected intraperitoneally with lithium-pilocarpine at 6-week-old with body weight of 180-220 g. Epilepsy model group (EP group), epilepsy + sodium hydrosulfide group (NaHS group). Morris water maze was used to detect spatial learning and memory in rats. Real-time fluorescence quantitative PCR was used to detect the expression of cystathionine-β-synthase (CBS) mRNA and CREB mRNA in the hippocampus. Results Compared with NC group, the learning and memory abilities of EP group were significantly decreased compared with NC group (P <0.01). The effective time of exploration in the third quadrant was significantly shortened (P <0.05). The third quadrant Swimming distance as a percentage of the total distance and crossing the platform significantly decreased (P <0.01); NaHS can improve the learning and memory ability of epilepsy rats, compared with the EP group, the spatial learning and memory ability of the test indicators were significantly different (P < 0.05, P <0.01). The level of CBS mRNA in hippocampus in EP group was significantly lower than that in NC group at 7 and 30 days (P <0.05), while the level of CBS mRNA in NaHS group was significantly higher than that in EP group (P <0.05) The expression of CREB mRNA in NaHS group was significantly higher than that in EP group at 24 h, 7 and 30 d (P <0.05), compared with NC group, There was no difference in d (P> 0.05). Conclusion Exogenous H2S may improve the learning and memory ability of epileptic rats by up-regulating the expression of CBS and CREB mRNA in the hippocampus of epileptic rats.