目的:探讨天麻素对氯化锂-匹罗卡品诱导癫痫大鼠模型中大脑皮质神经元凋亡因子表达的作用及其机制,为临床应用天麻素治疗癫痫提供分子生物学依据。方法:80只雄性SD大鼠随机分为5组:即对照组、模型组、60ms/kg天麻素预治疗组、120ms/kg天麻素预治疗组和180ms/kg天麻素预治疗组。采用氯化锂.匹罗卡品联合制备癫痫模型,治疗组给予不同剂量的天麻素预处理,模型组给以相同剂量的生理盐水。按Racine分级标准观察行为学变化,记录大发作潜伏期时间,于注射匹罗卡品后2h给予安定终止;3d后取材,免疫组织化学染色、免疫荧光染色和Western Blot检测凋亡相关蛋白表达变化。结果:天麻素治疗组较模型组,癫痫发作潜伏期延长,持续时间明显减少,程度减轻,死亡率显著降低,Bcl-2表达增加,Caspase-3显著降低。结论:天麻素可减少癫痫模型大鼠发作持续时间和程度,降低癫痫大鼠死亡率;天麻素通过增加癫痫大鼠大脑皮质Bcl-2和降低Caspase-3的表达而抑制凋亡,发挥脑保护作用。
Objective: To explore the protective effects of gastrodin on the expression of apoptosis factor in cortical neuron of the epileptic rats induced by lithium chloride-pilocarpine and the mechanisms and to provide the molecular basis whereby gastrodin treat the epilepsy in clinic. Methods: Eighty rats were randomly divided into five groups: control group, model group and treatment groups pretreated by gastrodin with 60 mg/kg, 120 mg/kg and 180 mg/kg. Epilepsy models were established by injecting lithium ehloride-pilocarpine. The treatment groups were given different dosages of gastrodin and model group was injected with same amount of saline. The behavior changes were observed according to the Racine Grading Standards. The latent period of grand seizure was recorded. The seizure was ended with valium in two hours after pilocarpine injection. Three days later, the expression of apoptosis related proteins were determined by immu- nohistochemistry, immunofluorescence and Western Blot. Results : Compared with model group, the latent period of treatment groups for epileptic attack was significantly prolonged, while the duration was shortened and mortality rate was decreased. The expression of Bcl-2 increased and Caspase-3 decreased markedly. Conclusion: The results indicate that gastrodin can reduce the seizure duration, severity, and the morality rate of epleptic rats. Gastrodin may protect the brain cells by increasing the expression of Bcl-2 and decreasing the expression of the Caspase-3.