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目的在建立氯化锂-毛果芸香碱癫持续状态模型的基础上,观察内质网应激相关因子CCAAT/增强子结合蛋白同源蛋白(CHOP)和Caspase-12的表达变化,探讨促红细胞生成素(EPO)产生脑保护作用的可能机制。方法 21～30 d SD大鼠96只随机分为对照组(n=32)、氯化锂-毛果芸香碱癫组(n=32)及EPO干预组(n=32),每组按6 h、24 h、48 h、72 h时间点又分为4个亚组,每亚组8只。观察各组大鼠的行为学变化,用免疫组织化学法检测各时间点各组大鼠CHOP及Caspase-12的表达水平。结果氯化锂-毛果芸香碱癫组大鼠海马区CHOP的表达水平明显增加,6 h开始增加,24 h达到高峰,后逐渐下降,至72 h仍高于对照组,与对照组同一时间点比较差异均有统计学意义(Pa<0.05);氯化锂-毛果芸香碱癫组大鼠海马区Caspase-12的表达水平在6 h开始增加,48 h达到高峰,72 h开始下降,但仍高于对照组,与对照组同一时间点比较差异均有统计学意义(Pa<0.05)。EPO干预组海马区CHOP及Caspase-12的表达较同一时间点氯化锂-毛果芸香碱癫组均下降,差异有统计学意义(Pa<0.05)。结论CHOP和Capase-12在癫发作后表达明显增加,提示内质网应激机制在癫性脑损伤的发生发展中具有重要作用;EPO可能通过内质网应激机制产生脑保护作用。
OBJECTIVE: To establish a continuous model of epilepticus of lithium chloride-pilocarpine, and to observe the changes of endoplasmic reticulum stress-related factors CCAAT / enhancer binding protein homologues (CHOP) and Caspase-12, and to explore the relationship between erythropoietin (EPO) to produce a possible mechanism of brain protection. Methods Ninety-six SD rats aged 21-30 days were randomly divided into control group (n = 32), lithium chloride-pilocarpine epilepsy group (n = 32) and EPO intervention group (n = 32) 24 h, 48 h, 72 h time point is divided into 4 subgroups, each subgroup of 8. The behavioral changes of rats in each group were observed. The expressions of CHOP and Caspase-12 in each group were detected by immunohistochemical method. Results CHOP expression in hippocampus of lithium chloride-pilocarpine epilepsy group increased significantly at 6 h, peaked at 24 h, then decreased gradually and remained higher at 72 h compared with the control group at the same time point (P <0.05). The expression level of Caspase-12 in hippocampus of lithium chloride-pilocarpine epilepsy group began to increase at 6 h, peaked at 48 h, but decreased at 72 h, but still higher than The difference between the control group and the control group at the same time point was statistically significant (Pa <0.05). Compared with the lithium chloride-pilocarpine epilepsy group, the expression of CHOP and Caspase-12 in the hippocampus of EPO-treated group were significantly decreased (P <0.05). Conclusion The expression of CHOP and Capase-12 in epileptic seizures increased significantly, suggesting that endoplasmic reticulum stress mechanism plays an important role in the development of epileptic brain injury. EPO may exert cerebral protective effects through the endoplasmic reticulum stress mechanism.