胶质细胞源性神经营养因子对培养脊髓运动神经元的影响(英文)  

Influence of glial cell line-derived neurotrophic factors on spinal cord motor neurons in vitro

作　　者：宋学琴[1] 李春岩[1] 王丽琴[1] 刘瑞春[1] 王晓娟[1] 

机构地区：[1]河北医科大学第二医院神经内科,河北省石家庄市050000

出　　处：《中国临床康复》2006年第5期147-149,i0006,共4页Chinese Journal of Clinical Rehabilitation

摘　　要：背景:胶质细胞源性神经营养因子对运动神经元具有营养作用,但不同浓度的营养因子对培养运动神经元体外生长影响的作用缺乏量化数据。目的:采用体外培养胚胎大鼠的脊髓运动神经元,观察不同浓度胶质细胞源性神经营养因子对其生长活性的作用。设计:以体外培养细胞为对象的验证性观察。单位:河北医科大学第二医院神经内科。材料:实验于2001-01/2002-09在河北医科大学第二医院神经内科实验室完成。选择成年清洁级雌雄SD大鼠合笼,取孕15d的大鼠胚胎进行脊髓分离。方法:取孕鼠15d胚胎的脊髓腹侧半组织进行原代体外分离培养。胶质细胞源性神经营养因子组按浓度1,10,50,100μg/L分为4组。对照组为不含胶质细胞源性神经营养因子的培养液。各组设立8个复孔,共做2个批次。从细胞形态学及应用MTT法观察不同浓度胶质细胞源性神经营养因子对大鼠脊髓运动神经元的影响。主要观察指标:培养运动神经元的细胞存活数目和细胞突起的长度。结果:①脊髓运动神经元细胞突起的长度比较:胶质细胞源性神经营养因子1μg/L组、10μg/L组、50μg/L组和100μg/L组明显高于对照组[(107.4±35.4068,160.5±38.5649,450.5±60.6403,293.5±67.3814,82.8±7.9725)μm,t=2.610-2.647,P<0.01]。②细胞存活数:胶质细胞源性神经营养因子1μg/L组、10μg/L组、50μg/L组和100μg/L组明显高于对照组[(13.9±0.8999,16.1±0.6680,20.1±0.6679,26.0±0.6030,10.5±0.7820)μm,t=2.211-2.312,P<0.05]。③MTT比色微量分析:胶质细胞源性神经营养因子1μg/L组、10μg/L组、50μg/L组和100μg/L组明显高于对照组[(0.350±0.0598,0.3667±0.0719,0.3819±0.0638,0.3953±0.0605,0.2858±0.0325)μm,t=2.259-2.577,P<0.05]。结论:不同浓度胶质细胞源性神经营养因子对体外培养大鼠胚胎脊髓运动神经元有不同程度的促生长作用。BACKGROUND： Glial cell line-derived neurotrophie factor （GDNF） is characterized by its trnphic function on motor neurons, but there is still lack of quantitative data coneerning the influence of different concentrations of the neurotrophic factor on the growth of in vitro cultured motor OBJECTIVE： To observe the influence of GDNF on neuronal growth by observing fetus rat spinal cord motor neurons cultured in vitro. DESIGN： Verifying observation taking in vitro euhured eells as subjects. SETTING： Neurological Department of the Second Hospital Affiliated to Hebei Medical College. MATERIALS： This experiment was conducted in the laboratory of Neurulogieal Department, the Second Hospital Affiliated to Hebei Medical College, between January 2001 and September 2002. Adult male and female rats were raised together in the same cage, embryonic rats at 15 days of gestation were obtained for spinal cord separation. METHODS： Ventral spinal tissues were obtained from embryonic rats at 15 days of gestation for primary in vitro culture. They were divided into four groups according to the density of GDNF. namely 1. 10, 50 and 100μg/L GDNF groups, while the culture medium in control group did not contain GDNF. Neurons were cultured in 8 wells for each group, which was repeated for two batches. Then the influence of GDNF on spinal cord motor neurons was observed from the perspective of cell morphology with MTT method. MAIN OUTCOME MEASURES： The survival rate of motor neurons and the length of cell processes. RESULTS： ① The length of spinal cord motor neuronal processes： It was found obviously longer in GDNF 1 μg/L group, 10 μg/L group, 50 μg/L group and 100 μg/L group than in control group [（107.4±35.406 8, 160.5±38.564 9, 450.5±60.640 3, 293.5±67.381 4, 82.8±7.972 5） μm, t=2.610-2.647, P 〈 0.01]. ② Cell survival rate： It was higher in GDNF 1μg/L group, 10 μg/L group, 50 μg/L group and 100 μg/L group than in control group [（13.9±0.899 9, 16.1±0.668 0, 20.1±0.667 9, 26.0±0.603 0

关 键 词：脊髓 神经组织蛋白质类 运动神经元 

分 类 号：R651.2[医药卫生—外科学]