外源性神经生长因子对血肿灶周神经元再生的影响

Effect of exogenous nerve growth factor on neuron regeneration in perihematoma region

目的:观察神经生长因子作用下血肿灶周神经元再生状况。方法:实验于2003-03/2004-04在河北医科大学第二医院影像科分子影像学实验室完成。健康家犬21只,随机分为3组:神经生长因子组(n=9):注血后0.5h,将神经生长因子2000AU立体定向导入血肿灶周区。脑出血对照组(n=6):只注血,不注药。对照组(n=3):只进针,不注血和注药。3组动物进入实验程序后前3d进行BrdU标记,在脑出血后3,10,28d3个时间点进行激光共聚焦显微镜检测,观察血肿灶周BrdU荧光单标和BrdU-NSE及BrdU-GFAP荧光双标细胞的数目和所在位置(BrdU为新生神经元的标记物,NSE为成熟神经元的标记物,GFAP为成熟星形胶质细胞的标记物)。结果:实验动物21只均进入结果分析。①新生神经元数目:神经生长因子组3,10,28d时BrdU单标阳性细胞数多于脑出血对照组(2.31±0.24,9.52±1.87,4.43±0.56;0.12±0.14,3.85±1.87,1.41±0.32,P<0.05),10和28d时BrdU-NSE和BrdU-GFAP双标阳性细胞数明显多于脑出血对照组(BrdU-NSE:3.84±0.24和6.23±1.92,1.35±0.71和1.39±0.24;BrdU-GFAP:4.51±2.08和10.53±2.47,1.65±0.08和1.37±0.13,P<0.05);神经生长因子组10d时BrdU单标阳性细胞数最多,28d时BrdU-NSE和BrdU-GFAP双标阳性细胞数明显增多。②新生神经元分布:神经生长因子组双标细胞的分布多位于血肿靠近额叶皮质面,在皮质与皮质下移行区内可见双标细胞,而脑出血对照组极罕见。结论:外源性神经生长因子立体定向导入血肿灶周区,能够通过刺激额叶皮质内源性神经干细胞再生、迁徙和分化的机制,促进血肿灶周神经功能的修复。

AIM： To study the effect of exogenous nerve growth factor （NGF）neuron regeneration in perihematoma region. METHODS： The experiment was conducted in the Laboratory of Molecular Imaging of Department of Image, Second Hospital of Hebei Medical University between March 2003 to April 2004. Twenty-one healthy dogs were randomly divided into three groups： NGF group （n=9）： NGF （2000AU） were stereotaxial led into the perihematoma region at 0.5 hour after injection of blood. Intracerebral hemorrhage （ICH） group （n=6）： only blood was injected. Control group （n=3）： only needle was injected. BrdU were labeled in animals on the first three days of experiment, and then were detected with laser confocal microscope respectively at 3, 10 and 28 days after ICH. Number and distribution of cells labeled by BrdU single-fluorescence, BrdU-NSE and double-fluorescence in perihematoma region were observed （BrdU was the label of newly-born neuron, NSE was the label of mature neuron, and GFAP was the label of mature horizontal cells）. RESULTS： A total of 21 experimental animals were involved in the analysis of results. ①The number of newly born neuron： that of BrdU single-labeled positive cells in the NGF group on the 3^nd, 10^th, and 280^th days were significantly greater than those in the ICH group （2.31±0.24,9.52＋1.87, 4.43±0.56;0.12±0.14,3.85±1.87,1.41±0.32,P 〈 0.05）, and that of BrdU-NSE and BrdU-GFAP double-labeled positive cells in the NGF group on the 100^th and 280^th day were greater than those in the ICH group （BrdU-NSE： 3.84±0.24 and 6.23±1.92,1.35±0.7I and 1.39±0.24; BrdU-GFAP： 431±9.08 and 10.53±2.47,1.65±0.08 and 1.37±0.13, P 〈 0.05）. The number of BrdU single-labeled positive cells in the NGF group was the most on the 10^thday, and number of BrdU-NSE and BrdU-GFAP double-labeled positive cells in the NGF group was significantly increased on the 28^th day. ②The distribution of newly born neuron： That of double-labeled cells in the NGF group mainly located in the perihematoma region that near the cortex of frontal lobes. Double-labeled cells could be seen in migratory region between cortex and sub-cortex, whereas little can be seen in the ICH control group. CONCLUSION： The stereotaxial leading of exogenous NGF can accelerate the neurological functional recovery in perihematoma region by stimulating the regeneration, migration and differentiation of endogenous nerve stem cells.