PDF(1334 KB)
PDF(1334 KB)
PDF(1334 KB)
反复高热惊厥大鼠脑内脑源性神经营养因子表达及神经元凋亡分析
Expression of brain-derived neurotrophic factor and apoptosis in the brain of rats with repeated febrile seizures
目的：研究大鼠反复高热惊厥（FS）后脑源性神经营养因子（BDNF）的改变及神经细胞凋亡情况。方法：51只雄性SD大鼠按随机数字法随机分为正常对照组（NC组），高热对照组（HC组）和高热惊厥组（FS组），热水浴法建立FS模型。酶联免疫吸附试验（ELISA）测定海马匀浆中BDNF水平，免疫组化法检测BDNF在脑组织各区的表达，原位末端标记法（TUNEL）检测脑组织细胞凋亡，统计学处理采用SPSS13.0统计学软件处理。结果：ELISA 法测得的FS组海马BDNF水平（89.9±12.5 ng/g）明显高于NC组（54.4±18.9 ng/g）和HC组（64.1±15.0 ng/g）（P＜0.01），FS组各脑区BDNF阳性神经元OD值亦明显高于NC组和HC组（P＜0.01）；FS组脑组织各区凋亡指数（AI）均明显高于NC组与HC组（P＜0.01），且脑组织各区细胞凋亡程度与BDNF在脑组织各区的表达呈正相关（r=0.332，P＜0.05）。结论：反复FS后大鼠脑内BDNF表达明显增加，并且与神经细胞的凋亡相关。［中国当代儿科杂志，2010，12（5）：382-385］
OBJECTIVE: To study the changes of brain-derived neurotrophic factor (BDNF) following repeated febrile seizures in rats and its possible correlation with neurocyte apoptosis. METHODS: Fifty-one male Sprague-Dawley (SD) rats were randomly assigned to three groups: normal control (n=14), febrile seizure (FS, n=18), hyperthermia alone (n=19). Febrile seizures were induced by hot water bath. The level of BDNF in the hippocampal homogenate was measured using ELISA and the expression of BDNF in various brain regions was measured by immunohistochemistry. The neurocyte apoptosis of the brain was determined by TdT-mediated biotinylated-dUTP nick end labling (TUNEL). RESULTS: The level of BDNF in the hippocampus in the FS group（89.9±12.5 ng/g）was higher than that in the normal control group（54.4±18.9 ng/g）and in the hyperthermia alone group （64.1±15.0 ng/g） (P<0.01). The OD value of BDNF positive neurons in various brain regions of the FS group was significantly higher than that of the normal control group (P<0.01) and the hyperthermia alone group (P<0.01). The FS group had significantly higher apoptotic index in various brain regions than the normal control and the hyperthermia alone groups (P<0.01). There was a positive correlation between the expression of BDNF and the apoptotic index in various brain regions (r=0.332, P＜0.05）. CONCLUSIONS: BDNF expression in the brain increases following repeated febrile seizures in rats, and the increased BDNF expression is correlated with neurocyte apoptosis.［Chin J Contemp Pediatr, 2010, 12 (5):382-385］
Febrile seizure / Brain-derived neurotrophic factor / Apoptosis / Rats
［1］Nakajima H, Uchida K, Kobayashi S, Inukai T, Horiuchi Y, Yayama T, et al. Rescue of rat anterior horn neurons after spinal cord injury by retrograde transfection of adenovirus vector carrying brain-derived neurotrophic factor gene［J］. Neurotrauma, 2007, 24（4）：703-712.
［2］Murashima YL, Suzuki J, Yoshii M. Developmental program of epileptogenesis in the brain of EL mice［J］. Epilepsia, 2005, 46（5）：10-16.
［3］周戬平，王帆，李瑞林，黄绍平，郭亚乐. 热性惊厥大鼠海马区c-Fos蛋白表达及苔藓纤维发芽特征［J］. 实用儿科临床杂志，2006，21（2）：101-103.
［4］郑飞霞，李光乾. 反复热性惊厥大鼠体内皮质醇水平变化［J］. 实用儿科临床杂志，2006，21（18）：1250-1253.
［5］Tejada S, Roca C, Sureda A, Rial RV, Gamundí A, Esteban S. Antioxidant response analysis in the brain after pilocarpine treatments［J］. Brain Res Bull， 2006，6（9）：587-592.
［6］Kawakami Y, Monobe M, Kuwabara K, Fujita T, Maeda M, Fujino O, et al. A comparative study of nitric oxide, glutathione, and glutathione peroxidase activities in cerebrospinal fluid from children with convulsive diseases/children with aseptic meningitis［J］. Brain Dev, 2006, 2（8）：243-246.
［7］Tongiorgi E, Domenici L, Simonato M. What is the biological significance of BDNF mRNA targeting in the dendrites? Clues from epilepsy and cortical development［J］. Mol Neurobiol, 2006, 33（1）：17-32.
［8］Shetty AK, Zaman V, Shetty GA. Hippocampal neurotrophin levels in a kainate model of temporal lobe epilepsy：a lack of correlation between brain-derived neurotrophic factor content and progression of aberrant dentate mossy fiber sprouting［J］. Neurochem, 2003, 87（1）：147-159.
［9］Hattiangady B, Rao MS, Zaman V, Shetty AK. Incorporation of embryonic CA3 cell grafts into the adult hippocampus at 4-months after injury：effects of combined neurotrophic supplementation and caspase inhibition［J］. Neuroscience, 2006, 139（4）：1369-1383.
［10］Hagihara H, Hara M, Tsunekawa K, Nakagawa Y, Sawada M, Nakano K.Tonic-clonic seizures induce division of neuronal progenitor cells with concomitant changes in expression of neurotrophic factors in the brain of pilocarpine treated mice［J］. Brain Res Mol Brain Res, 2005, 139（2）：258-263.
［11］王虔，程焱，吴伟，梁浩，张瑞珠. 脑源性神经营养因子对谷氨酸损伤神经元保护作用的研究［J］. 中华老年心脑血管病杂志，2006，8（1）：57-60.
［12］Kim YH, Rhyu IJ, Park KW, Eun BL, Kim YI, Rha HK, et al. The induction of BDNF and c-fos mRNA in the hippocampal formation after febrile seizures［J］. Neuroreport，2001，12（15）：3243-3246.
［13］Wang CX, Song JH, Song DK, Yong VW, Shuaib A, Hao C. Cyclin-dependent kinase-5 prevents neuronal apoptosis through ERK-mediated upregulation of Bcl-2［J］. Cell Death Differ, 2006, 13（7）：1203-1212.
［14］Rose CR, Blum R, Kafitz KW, Kovalchuk Y, Konnerth A. From modulator to mediator：rapid effects of BDNF on ion channels［J］. Bioessays，2004，26（11）：1185-1194.
［15］D'Intino G, Vaccari F, Sivilia S, Scagliarini A, Gandini G, Giardino L, et al. A molecular study of hippocampus in dogs with convulsion during canine distemper virus encephalitis［J］. Brain Res, 2006, 1098（1）：186-195.
