神经生长因子对难治性癫癎儿童脑海马癎性放电区神经细胞的影响

方琼; 陈燕惠

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中国当代儿科杂志 ›› 2011, Vol. 13 ›› Issue (3) : 236-239.

论著·实验研究

神经生长因子对难治性癫癎儿童脑海马癎性放电区神经细胞的影响

方琼，陈燕惠

作者信息 +

Effects of nerve growth factor on neural cells in the epileptiform discharge region of the hippocampus in children with intractable epilepsy

FANG Qiong, CHEN Yan-Hui

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文章历史 +

摘要

目的：探讨神经生长因子（NGF）对难治性癫癎儿童海马癎性放电区星形胶质细胞和神经元的影响。方法：对术中取下的癫癎儿童脑海马癎性放电区组织块，采用解离细胞即刻培养技术，在空白对照组及NGF浓度分别为12.5、50、100 ng/mL 的条件下进行培养，通过免疫荧光细胞化学法，以双苯酰亚胺（Bb）染核，联合细胞特异性标志蛋白GFAP、MAP2分别标记星形胶质细胞、神经元，在倒置荧光显微镜下计数神经细胞总数，并计算相应的阳性细胞比率。结果：NGF 作用 4 h 后，海马区 GFAP 阳性的星型胶质细胞比率及 MAP2 阳性神经元细胞比率较空白对照组升高，并随着 NGF 浓度的增高而增加（P<0.05）。结论：NGF 对癫癎儿童损伤海马区的GFAP+ 星形胶质细胞和MAP2+神经元可能起到一定的保护作用。

Abstract

OBJECTIVE: To study the effects of nerve growth factor (NGF) on the astrocytes and neurons in the epileptiform discharge region of the hippocampus in children with intractable epilepsy. METHODS: Acutely dissociated cells from the epileptiform discharge region of the hippocampus were exposed to blank control group and NGF treatment groups (NGF concentrations: 12.5, 50 and 100 ng/mL). Astrocytes and neurons were identified by bisbenzimide staining combined with the specific makers such as GFAP and MAP2. The total number of neural cells was counted under an inversion fluorescence microscope. The percentage of immunostaining positive cells was calculated. RESULTS: After 4 hrs of culture, the percentages of GFAP+ astrocytes and MAP2+ neurons in the NGF treatment groups were higher than those in the blank control group, and increased with increasing NGF concentration (P<0.05). CONCLUSIONS: NGF may have protective effects on GFAP+ astrocytes and MAP2+ neurons in the injured hippocampus of children with epilepsy.

导出引用

方琼，陈燕惠. 神经生长因子对难治性癫癎儿童脑海马癎性放电区神经细胞的影响[J]. 中国当代儿科杂志. 2011, 13(3): 236-239

FANG Qiong, CHEN Yan-Hui. Effects of nerve growth factor on neural cells in the epileptiform discharge region of the hippocampus in children with intractable epilepsy[J]. Chinese Journal of Contemporary Pediatrics. 2011, 13(3): 236-239

中图分类号： R742.1

参考文献

［1］Thippeswamy T, McKay JS, Quinn J, Morris R. Either nitric oxide or nerve growth factor is required for dorsal root ganglion neurons to survive during embryonic and neonatal development［J］. Brain Res Dev Brain Res, 2005, 154(2): 153-164.

［2］Ozdinler PH, Ulupinar E, Erzurumlu RS. Dose and age-dependent axonal responses of embryonic trigeminal neurons to localized NGF via p75NTR receptor［J］. J Neurobiol, 2005, 62(2): 189-206.

［3］吴立文.难治性癫痫的诊断和治疗［J］. 中华神经科杂志，2002，35（1）：55-57.

［4］陈燕惠，张燕舞，吕敏. 妊娠 20～24 周人脑脑室管膜下区神经细胞组成［J］. 解剖学杂志，2007，30（5）：572-575.

［5］Tassic L, Colombo N, Garbelli R, Francione S, Lo Russo G, Mai R. Focal cortical dysplasia: neuropathological subtypes, EEG, neuroimaging and surgical outcome［J］. Brain, 2002, 125(8): 1719-1732.

［6］Takaya S, Hanakawa T, Hashikawa K, Ikeda A, Sawamoto N, Nagamine T, et al. Prefrontal hypofunction in patients with intractable mesial temporal lobe epilepsy［J］. Neurology, 2006, 67(9): 1674-1676.

［7］Szyndler J, Piechal A, BlecharzKlin K, Skórzewska A, Maciejak P, Wallkowiak J, et al. Effect of kindled seizures on rat behavior in water Morris maze test and amino acid concent rations in brain structures［J］. Pharmacol Rep, 2006, 58(1):75-82.

［8］Kotloski R, Lynch M, Lauersdorf S, Sutula T. Repeated brief seizures induce progressive hippocampal neuron loss and memory deficits［J］. Prog Brain Res, 2002, 135: 95-110.

［9］Yu LM, Miller FD, Schoichet MS. The use of immobillized neurotrophins to support neuron survival and guide nerve fiber growth in compartmentalized chambers［J］. Biomaterials, 2010, 31(27): 6987-6999.

［10］Davis-Lopez de Carrizosa MA, Morado-Diaz CJ, Morcuende S, de la Cruz RR, Pastor AM. Nerve growth factor regulates the firing patterns and synaptic composition of motoneurons［J］. J Neurosci, 2010, 30(24): 8308-8319.

［11］丛树艳，刘冰，柳忠兰. NGF对癫癎大鼠海马神经细胞wtp53表达的影响［J］. 中风与神经疾病杂志，2000，14（4）：181-183.

［12］Hasegawa D, Matsuki N, Fujita M, Ono K, Orima H. Kinetics of glutamate and gamma-aminobutyric acid in cerebrospinal fluid in a canine model of complex partial status epilepticus induced by kainic acid［J］. J Vet Med Sci, 2004, 66(12):1555-1559.

［13］Pettigrew DB, Li YQ, Kuntz C, Crutcher KA. Global expression of NGF promotes sympathetic axonal growth in CNS white matter but does not alter its parallel orientation［J］. Exp Neurol, 2007, 203(1): 95-109.

［14］胡勇，邬建荣，孔元原，俞海国，汤云珍. 神经生长因子对新生大鼠缺氧缺血性脑损伤的保护作用［J］.中国当代儿科杂志，2001，3（2）：170-172.

［15］严超英，周文莉，徐乃军，武辉. 外周应用神经生长因子对新生大鼠缺氧缺血脑损伤的保护作用［J］. 中国新生儿科杂志，2007，22（2）：93-94.

［16］刘冰，柳忠兰，何桂芹. 神经生长因子对红藻氨酸诱导癫癎发作大鼠海马神经细胞凋亡的影响［J］. 中国神经免疫学和神经病学杂志，2000，7(1)：15-19.

［17］黄艳，孔庆兖，金淑静, 柳忠兰. 神经生长因子对大鼠癫癎脑损伤的保护作用［J］. 徐州医学院学报，2000, 20（3）: 192-196.

［18］李广宪，易晓燕，吕受卿，张湘玲，祁岩超. 神经生长因子治疗小儿癫癎［J］.实用儿科临床杂志. 1994，9(9): 94-96.

［19］David Y, Cacheaux LP, Lvens S, Lapilover E, Heinemann U, Kaufer D, et al. Astrocytic dysfunction in epileptogenesis: consequence of altered potassium and glutamate homeostasis［J］. J Neurosci, 2009, 29(34): 10588-10599.

［20］梁建民，崔新明，李秀杰，张淑琴. 颞叶癫癎大鼠海马CA1区超微结构［J］. 吉林大学学报（医学版），2005，31（5）：720-723.

［21］Du Y, Chen CP, Tseng CY, Eisenberg Y, Firestein BL. Astroglia-mediated effects of uric acid to protect spinal cord neurons from glutamate toxicity［J］. Glia, 2007, 55(5): 463-472.