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

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

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.