依达拉奉预处理对惊厥持续状态幼年大鼠海马神经元保护及IL-lβ表达的影响

王海萍; 李光乾

PDF(1618 KB)

中国当代儿科杂志 ›› 2010, Vol. 12 ›› Issue (3) : 205-210.

论著·实验研究

依达拉奉预处理对惊厥持续状态幼年大鼠海马神经元保护及IL-lβ表达的影响

王海萍，李光乾

作者信息 +

Protection of edravone on neurons and its effects on the expression of interleukin-lβ in juvenile rat hippocampus following status convulsion

WANG Hai-Ping, LI Guang-Qian

Author information +

文章历史 +

摘要

目的：观察依达拉奉预处理对幼年大鼠惊厥持续状态（SC）后脑组织海马神经元的保护作用及白介素-lβ（IL-lβ）表达的影响。方法：将195只Sprague-Dawley幼年雄性大鼠随机分为生理盐水对照组（NS组）、惊厥持续状态组（SC组）和依达拉奉干预组（ED组）；各组又均按时间点分为4、12、24、48、72 h 5个亚组。采用氯化锂-匹鲁卡品化学点燃法制备幼年大鼠SC模型。ED组于惊厥前3 d予以ED腹腔注射，每天1次，连续3 d。观察大鼠海马病理学改变及细胞凋亡以及IL-lβ蛋白表达情况。结果：电镜显示SC组惊厥后24 h海马少量神经元出现核固缩，大量内质网扩张；48 h以后内质网扩张较前有所减轻，但线粒体肿胀加重。ED组各时间点神经元改变均较SC组减轻。SC组在惊厥后12 h海马TUNEL阳性细胞数已显著高于NS 组，48 h达高峰，而ED组TUNEL阳性细胞数均较SC组显著下降，但仍高于NS组。免疫组化法示12 h、24 h、48 h、72 h SC组大鼠海马中IL-1β表达增强，与NS组比较差异有统计学意义；与SC组比较，ED组IL-1β表达明显降低，差异有统计学意义。结论：ED可下调匹鲁卡品致痫大鼠海马IL-1β的表达，减轻细胞凋亡，提示ED对SC引起的脑损伤有保护作用。［中国当代儿科杂志，2010，12（3）：205-210］

Abstract

OBJECTIVE: To study the possible protection of edaravone on neurons of the hippocampus after status convulsion (SC) and its effects on the expression of interleukin-1β (IL-lβ) in juvenile rats. METHODS: One hundred and ninety-five juvenile male Sprague-Dawley rats were randomly divided into three groups: SC, edaravone pretreatment and normal saline control (control group). Each group was subdivided into five groups sacrificed at 4, 12, 24, 48 and 72 hrs after SC induction. SC model was prepared using lithium-pilocarpine. The edaravone pretreatment group received edaravone by intraperitoneal injection once daily three days before convulsion induction. Histopathologic changes in the hippocampus were viewed under a light microscope and an electron microscope. Expression of apoptosis cells was observed by TdT-mediated dUTP nick end labeling (TUNEL). Expression of IL-lβ protein was determined by immunohistochemistry. RESULTS: Under the electron microscrope, a small quantity of neurons showed karyopycnosis and endocytoplasmic reticulum (ER) expanded remarkably 24 hrs after SC induction; at 48 hrs the ER expanding was alleviated somewhat but mitochomdria swelling was more severe. The edaravone pretreatment group showed less severe neuronal changes compared with the SC group under the microscopes. The TUNEL positive cells in the hippocampus of the SC group were significantly more than those of the control group 12 hrs, and peaked at 48 hrs after SC induction. The edaravone pretreatment group showed decreased TUNEL positive cells in the hippocampus compared with the SC group, although the positive cells were more than those in the control group between 12 and 48 hrs after SC induction. The immunohistochemistry assay demonstrated that the expression of IL-lβ in the hippocampus of the SC group increased significantly compared with that of the control group 12, 24, 48 and 72 hrs after SC induction. Edaravone pretreatment resulted in a significantly decreased IL-lβ expression in the hippocampus as compared with the SC group. CONCLUSIONS: Edaravone pretreatment may decrease the IL-1β expression and neuronal apoptosis in the hippocampus. This suggests that edaravone may have protective effects against the hippocampal damage caused by SC.［Chin J Contemp Pediatr, 2010, 12 (3):205-210］

导出引用

王海萍, 李光乾. 依达拉奉预处理对惊厥持续状态幼年大鼠海马神经元保护及IL-lβ表达的影响[J]. 中国当代儿科杂志. 2010, 12(3): 205-210

WANG Hai-Ping, LI Guang-Qian. Protection of edravone on neurons and its effects on the expression of interleukin-lβ in juvenile rat hippocampus following status convulsion[J]. Chinese Journal of Contemporary Pediatrics. 2010, 12(3): 205-210

中图分类号： R-33

参考文献

［1］Araújo IM, Gil JM, Carreira BP, Mohapel P, Petersen A, Pinheiro PS, et al. Calpain activation is involved in early caspase-independent neurodegeneration in the hippocampus following status epilepticus［J］. Neurochem, 2008, 105（3）：666-667.
［2］Vezzani A, Granata T. Brain inflammation in epilepsy：experimental and clinical evidence［J］. Epilepsia, 2005, 46（11）：1724-1743.
［3］王礼周，李树军，赵东菊，张贺，石太新，杨达胜，等.新生儿缺氧缺血性脑病血清半胱氨酸蛋白酶-1、白细胞介素-lβ、-18测定的意义［J］.实用儿科临床杂志，2005，20（2）：127-128.
［4］Yasuoka N, Nakajima W, Ishida A, Takada G. Neuroprotection of edaravone on hypoxic-ischemic brain injury in neonatal rats［J］. Brain Res Dev Brain Res，2004，151(1-2)：129-139.
［5］王海萍，邓小龙，李光乾. 依达拉奉对惊厥持续状态幼年大鼠海马IL-lβ、NF-κB表达及细胞凋亡的影响［J］.中华儿科杂志，2009，47（8）：575-580.
［6］André V, Dubé C, Francois J, Leroy C, Rigoulot MA, Roch C, et al.Pathogenesis and pharmacology of epilepsy in the lithium-pilocarpine model［J］.Epilepsia, 2007, 48（Suppl 5）：41-47.
［7］Dinarello CA.Immunological and inflammatory functions of the interleukin-1 family［J］. Annu Rev Immunol, 2009, 27：519-550.
［8］Li B, Mahmood A, Lu O, Wu H, Xiong Y, Qu C, et al. Simuastatin attenuates microglial cells and astrocyte activation and decreases interleukin-1 beta level after traumatic brain injury［J］. Neurosurgery, 2009, 65(1): 179-185.
［9］Hailer NP, Vogt C, Korf HW, Dehghani F. Interleukin-1beta exacerbates and interleukin-1 receptor antagonist attenuates neuronal injury and microglial activation after excitotoxic damage in organotypic hippocampal slice cultures［J］. Eur J Neurosci, 2005, 21（9）：2347-2360.
［10］Sayyah M, Beheshti S, Shokrgozar MA, Eslami-far A, Deljoo Z, Khabiri AR, et al. Antiepileptogenic and anticonvulsant activity of interleukin-1 beta in amygdala-kindled rats［J］. Exp Neuro, 2005, 191（1）：145-153.
［11］毛定安, 刘利群, 薄涛, 刘洁明，白海涛，熊洁. 幼年大鼠惊厥性脑损伤白细胞介素-1β 和白细胞介素-18 的表达及川弓嗪干预的影响［J］. 中国急救医学, 2007, 27（1）：42-45.
［12］De Simoni MG, Perego C, Ravizza T, Moneta D, Conti M, Marchesi F, et al. Inflammatory cytokines and related genes are induced in the rat hippocampus by limbic status epilepticus［J］. Eur J Neurosci, 2000, 12（7）：2623-2633.
［13］Patel HC, Ross FM, Heenan LE. Neurodegenerative actions of interleukin-1 in the rat brain are mediated through increases in seizure activity［J］. J Neurosci Res, 2006, 83（3）：385-391.
［14］宋贵军, 唐建武, 尹琳, 王乃昌, 包礼平.依达拉奉对脑缺血-再灌注损伤大鼠的抗细胞凋亡与神经保护作用［J］. 医药导报, 2007, 26（6）：582-585.
［15］Martinian L, Boer K, Middeldorp J, Hol EM, Sisodiya SM, Squier W, et al. Expression patterns of glial fibrillary acidic protein (GFAP)-delta in epilepsy-associated lesional pathologies［J］. Neuropathol Appl Neurobiol, 2009, 35（4）：394-405.
［16］Sama MA, Mathis DM, Furman JL, Abdul HM, Artiushin IA, Kraner SD, et al. Interleukin-1beta-dependent signalin g between astrocytes and neurons depends critically on astrocytic calcineurin/NFAT activity［J］. J Biol Chem, 2008, 283（32）：21953-21964.