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目的 :红藻氨酸 (kainicacid ,KA)诱导的大鼠边缘叶发作模型 ,检测半胱氨酸天冬酶 9(caspase 9)在癫大鼠海马神经元表达。方法 :立体定位杏仁核注射KA诱导癫发作 ,以持续记录脑电、局部脑血流 (regionalcerebralbloodflow ,r CBF) ,用TUNEL染色和cresylviolet染色观察海马神经元存活和凋亡 ;用免疫荧光和Westernblot检测海马caspase 9的表达。结果 :发作终止 4h时caspase 9出现裂解片段 ,8h时出现TUNEL阳性细胞 ,2 4h时达高峰。脑室内注射caspase 9抑制剂z LEHD fluoromethylketone(z LEHD fmk)可减少TUNEL阳性细胞 ,增加存活神经元 ;发作后在KA注射同侧海马的CA3区神经元caspase 9免疫反应性增强 ;对侧海马未见TUNEL阳性细胞及caspase 9的上述变化。发作前后r CBF无明显变化。结论 :癫发作可诱导caspase 9的激活。caspase 9可能是癫潜在的治疗靶点。
OBJECTIVE: The rat model of limbic lobule induced by kainic acid (KA) was used to detect the expression of caspase 9 in the hippocampal neurons of epileptic rats. Methods: Epileptic seizures were induced by stereotactic injection of KA into the amygdaloid nucleus to continually record regional cerebral blood flow (rCBF). TUNEL staining and cresylviolet staining were used to observe the survival and apoptosis of hippocampal neurons. Immunofluorescence and Western blot The expression of caspase 9 in hippocampus was detected. Results: The cleaved fragment of caspase 9 appeared at 4h after the onset of seizures. TUNEL positive cells appeared at 8h and peaked at 24 h. Intracerebroventricular injection of caspase 9 inhibitor z LEHD fluoromethylketone (z LEHD fmk) decreased TUNEL-positive cells and increased viable neurons; caspase 9 immunoreactivity in CA3 neurons of KA injected ipsilateral hippocampus increased after the onset of seizures; contralateral hippocampus See the above changes of TUNEL positive cells and caspase 9. Before and after the onset of r CBF no significant change. Conclusion: Epileptic seizures induce caspase 9 activation. Caspase 9 may be a potential therapeutic target for epilepsy.