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背景:近年来,许多学者对癫痫与免疫的关系从细胞和分子水平进行了研究,发现癫痫患者存在细胞、分子及其功能的多种免疫学改变。目的:观察实验性癫痫大鼠海马神经元的变性,以及小胶质细胞MHC-Ⅰ、MHC-Ⅱ和C3b受体的表达。设计:随机对照动物实验。单位:大连大学医学院;日本三重大学。材料:实验于2001/2002在吉林北华大学免疫学实验室完成。取成年Wistar大鼠40只,随机分为模型组和对照组两组,每组20只。方法:模型组大鼠皮下注射海仁酸(12mg/kg)建立实验性癫痫大鼠模型,对照组不干预,观察模型组大鼠注射海仁酸6h期间湿狗样变行为变化。给药后3d处死大鼠,采用结晶紫染色方法观察实验性癫痫大鼠海马神经元的变性;采用免疫组织化学染色方法观察实验性癫痫大鼠MHC分子和补体C3b受体的表达。主要观察指标:两组大鼠大脑海马神经元变性,MHC-Ⅰ类分子、MHC-Ⅱ类分子和C3b受体的密度比较。结果:40只大鼠全部进入结果分析。①模型组大鼠注射海仁酸3h以内出现湿狗样变及抽搐发生,在第3～6小时出现更为剧烈的癫痫持续状态。②模型组大鼠大脑海马锥状细胞层可见到神经元的变性,对照组未见神经元损伤。③模型组大鼠大脑海马MHC-Ⅰ类分子、MHC-Ⅱ类分子和C3b受体的密度分别为(201.6±6.43),(493.8±7.92)和(362.5±3.18)个/视野,对照组未见明显表达,两组比较差异显著(P<0.01)。结论:①皮下注射海仁酸可成功建立实验性癫痫大鼠模型。②实验性癫痫大鼠大脑海马硬化发生过程中,存在着补体系统参与的免疫炎症反应,说明癫痫的免疫炎症机制。
Background: In recent years, many scholars have studied the relationship between epilepsy and immunity at the cellular and molecular level and found that there are many immunological changes in cells, molecules and their functions in patients with epilepsy. Objective: To observe the degeneration of hippocampal neurons and the expression of MHC-Ⅰ, MHC-Ⅱ and C3b receptors in microglia in experimental epilepsy rats. Design: Randomized controlled animal experiments. Unit: Dalian University School of Medicine; Japan’s Mie University. Materials: The experiment was performed in Immunology Laboratory, Beihua University, Jilin Province from 2001 to 2002. Forty adult Wistar rats were randomly divided into model group and control group, with 20 rats in each group. Methods: Experimental rats with epilepsy were injected subcutaneously with naringin (12mg / kg) in the model group. The rats in the control group were not intervened. The behavior of wet dog in the model group was observed during 6h injection. The rats were sacrificed 3 days after the administration, and the degeneration of hippocampal neurons in experimental epilepsy rats was observed by crystal violet staining. The expression of MHC molecules and complement C3b receptor in experimental epileptic rats was observed by immunohistochemical staining. MAIN OUTCOME MEASURES: Degeneration of hippocampal neurons, comparison of density of MHC class I molecules, MHC class II molecules and C3b receptors in two groups of rats. Results: All 40 rats entered the result analysis. ① In the model group, wet dog-like changes and convulsions occurred within 3 hours after injection of meric acid, and more severe status epilepticus occurred in the third to sixth hours. ② In the model group, degeneration of neurons was observed in the hippocampal pyramidal cells of rats in the model group, and no neuronal injury was found in the control group. ③ The density of MHC-I molecules, MHC-II molecules and C3b receptors in the hippocampus of the model group were (201.6 ± 6.43), (493.8 ± 7.92) and (362.5 ± 3.18) / field, respectively, See the obvious expression, the difference between the two groups was significant (P <0.01). Conclusion: ① Subcutaneous injection of meric acid can successfully establish experimental epilepsy rat model. (2) During the process of hippocampal sclerosis in experimental epilepsy rats, immune and inflammatory reactions involved in the complement system exist, indicating the immune and inflammatory mechanisms of epilepsy.