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目的立体定向手术建立海人酸(KA)颞叶癫痫大鼠模型,检测海马内IL-1β、TNF-α蛋白及其mRNA的表达。方法大鼠随机分为空白对照组、生理盐水对照组和模型组。模型组大鼠一侧海马CA3区注射KA(生理盐水组注射生理盐水),观察其行为学特征,HE染色和Nissl染色以及电镜观察其病理学改变,免疫组化法检测大鼠海马内IL-1β、TNF-α蛋白的表达,原位杂交法检测TNF-αmRNA的动态表达。结果大鼠注射KA后出现湿狗样抖动、头面部肌阵挛、肢体阵挛及全面强直阵挛发作等,病理结果显示海马神经元变性、缺失及胶质细胞增生,模型组IL-1β在致痫后36、h表达水平明显增加并于12 h达高峰,之后逐渐下降,7 d后与对照组相比差异无统计学意义(P>0.05);TNF-α蛋白与mRNA表达时程基本一致,3 h出现,12 h达高峰,而后逐渐下降,7 d后回归至对照组表达水平,15、30 d又高于对照组(P<0.05)。结论(1)大鼠一侧海马注射KA是人类颞叶癫痫理想的动物模型;(2)内源性IL-1β、TNF-α参与了癫痫发病机制。
OBJECTIVE: To establish a model of temporal lobe epilepsy rat model of kainate acid (KA) by stereotactic surgery, and to detect the expression of IL-1β, TNF-α and its mRNA in hippocampus. Methods The rats were randomly divided into blank control group, normal saline control group and model group. The rats in the untreated group were injected KA into the hippocampus CA3 area of the hippocampus to observe their behavioral characteristics. The pathological changes were observed by HE staining and Nissl staining and electron microscopy. The levels of IL- 1β, TNF-α protein expression, in situ hybridization assay of TNF-αmRNA expression. Results After the KA injection, wet dog-like jitter, head and facial myoclonus, limb clonus and generalized tonic clonic seizures were observed. The pathological results showed degeneration and deletion of hippocampal neurons and glial cell proliferation. The levels of IL-1β The expression of TNF-α at 36 and h after epileptic seizure increased significantly and peaked at 12 h, then gradually decreased. There was no significant difference between the control group and the 7 d after operation (P> 0.05) (P <0.05), and reached the peak at 3 h and peaked at 12 h, then decreased gradually and returned to the control group after 7 days. Conclusions (1) KA injection into rat hippocampus is an ideal model of human temporal lobe epilepsy. (2) Endogenous IL-1β and TNF-α are involved in the pathogenesis of epilepsy.