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目的 探讨强脑因子对大鼠胎鼠脑神经细胞培养物的生物学保护效应 方法 取16日龄Wistar大鼠脑神经细胞经体外培养后,分别进行(1)组织形态学观察:在神经细胞培养物中加入不同剂量强脑因子(1 10和100mg/L),光学显微镜下观察细胞形态学变化 在电子显微镜下观察神经细胞超微结构变比;(2)神经细胞代谢活性检测:应用四唑盐(tetrazolium,MTT)法测定不同剂量强脑因子对神经细胞代谢活性的影响;(3)可溶性蛋白质水平测定:应用Bradford蛋白定量法测定不同剂量强脑因子对神经细胞胞浆内可溶性蛋白质水平的影响;(4)强脑因子抗神经细胞凋亡试验:应用神经元特异性烯醇化酶(neuron-specific enolase,NSE)定量法,测定强脑因子与奥地利产脑活素对NSE活性表达的影响。结果(1)强脑因子可促进神经细胞分化成熟 突起增多并延长、细胞数量增加。(2)在不同剂量强脑因子作用下 神经细胞MTT代谢率增强,细胞浆内蛋白质水平升高 而且随着强脑因子剂量的增加均呈现出明显的剂量依赖关系;促进NSE表达活性,利于神经母细胞分化(3)强脑因子可增强神经细胞抗缺氧作用及抗谷氨酸所致的细胞凋亡效应 且功效强于奥地利产脑活素 结论 在相同实验条件下,强脑因子对体外培养的神经细胞有生物学保护作用,效果优于脑活素
Objective To investigate the biological protective effect of Intracerebroventricular factor on cultured rat fetal brain neurons.Methods Cultured 16-day-old Wistar rat brain neurons were cultured in vitro and respectively (1) Morphological observation: (1 10 and 100 mg / L) were added into the medium to observe the morphological changes of the cells under the light microscope. The ultrastructural changes of the nerve cells were observed under an electron microscope. (2) Detection of the metabolic activity of nerve cells: (3) Determination of soluble protein levels: Bradford protein quantification method was used to determine the effect of different doses of strong brain-derived factor on the level of soluble protein in the cytoplasm of nerve cells (4) Anti-Nerve Cell Apoptosis Test: Neuron-specific enolase (NSE) assay was used to determine the effect of strong brain natriuretic peptide and austenite on NSE activity . Results (1) Strong brain-derived factor can promote the differentiation and maturation of nerve cells increased and prolonged, the number of cells increased. (2) Under the condition of different dose of strong brain factor, the MTT metabolic rate of nerve cells increased, the level of protein in cytoplasm increased, and showed a dose-dependent relationship with the increase of dose of strong brain factor; promoted NSE expression activity, (3) Strong brain-derived factor enhances neuronal anti-anoxia effect and anti-glutamate-induced apoptosis and its efficacy is better than Austrian-produced cerebrolysin Conclusion Under the same experimental conditions, Cultured nerve cells have a biological protective effect, the effect is better than cerebrolysin