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目的:采用立体定向手术的方法建立海人酸颞叶癫痫大鼠模型,探讨其行为学表现,并观察海马各区病理学改变以及肿瘤坏死因子α的表达特点。方法:实验于2005-03/09在哈尔滨医科大学附属第二医院实验动物中心进行。①分组:健康雄性清洁级Wistar大鼠104只,分为空白对照组8只、生理盐水对照组32只和模型组64只,后两组每组又分3,6,12,24h,3,7,15,30d等8个时点,其中生理盐水对照组每时点4只,模型组每时点8只。②大鼠一侧海马CA3区注射海人酸1.0μL(生理盐水对照组抽取生理盐水1.0μL),缓慢注射,于10～15min内注射完毕,留针5min后拔出。③观察各组大鼠的行为学表现,苏木精-伊红染色法观察海马各区病理学改变,免疫组化法检测大鼠脑海马内肿瘤坏死因子α蛋白的动态表达。结果:①在行为学上,模型组大鼠注射海人酸后可出现湿狗样抖动、头面部肌肉阵挛、肢体阵挛及全面强直阵挛发作。空白对照组和生理盐水对照组未见癫痫发作。②病理学上,一侧海马注射可造成双侧损害,注射侧重以CA3和门区为主,对侧轻且以CA1和门区为主。③免疫组化结果:肿瘤坏死因子α蛋白在海马各区均有表达,神经元、胶质细胞均见表达。海马内肿瘤坏死因子α蛋白在3h出现,12h时达高峰,而后逐渐下降,7d后与生理盐水对照组表达水平相似,15,30d又稍高于生理盐水对照组。结论:大鼠一侧海马注射海人酸是理想的人类颞叶癫痫动物模型,内源性肿瘤坏死因子α可能通过各种方式影响脑内功能活动,参与癫痫发病过程。
OBJECTIVE: To establish a rat model of kainic acid temporal lobe epilepsy by stereotactic surgery and investigate its behavior and observe the pathological changes and the expression of tumor necrosis factor-α in the hippocampus. Methods: The experiment was performed at Experimental Animal Center of the Second Affiliated Hospital of Harbin Medical University from March to September in 2005. ① Grouping: 104 healthy male Wistar rats were divided into blank control group (n = 8), saline control group (n = 32) and model group (n = 64). The latter two groups were divided into 3, 6, 12, 24 h, 7, 15, 30d and other 8 time points, including saline control group at each time point 4, the model group at each time point 8. ② The hippocampal CA3 area of the rat was injected with 1.0 μL of kainic acid (physiological saline control group, 1.0 μL of normal saline) and injected slowly. The injection was completed within 10-15 min and the needle was withdrawn after 5 min. ③ The behavior of rats in each group was observed. The pathological changes in hippocampus were observed by hematoxylin-eosin staining. The dynamic expression of tumor necrosis factor-α protein in hippocampus was detected by immunohistochemistry. Results: ① In behavioral studies, wet dog-like jitter, head and face muscular clonus, limb clonus and generalized tonic clonic seizures were observed in the model rats after the injection of kainic acid. No control group and saline control group seizures. ② pathology, bilateral hippocampal injection can cause bilateral damage, focusing on the injection of CA3 and door-based, contralateral to light and CA1 and door-based. ③ Immunohistochemistry results: Tumor necrosis factor alpha protein expressed in all regions of the hippocampus, neurons, glial cells were seen. The level of tumor necrosis factor alpha protein in hippocampus appeared at 3h and peaked at 12h, then gradually decreased. After 7d, the expression of tumor necrosis factor alpha protein was similar to that in normal saline control group, and slightly higher than saline control group on the 15th and 30th days. Conclusion: The injection of kainate into rat hippocampus is an ideal animal model of human temporal lobe epilepsy. Endogenous tumor necrosis factor-α may affect brain functional activity in various ways and participate in the process of epilepsy.