强脑因子生物学特性的实验研究

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

Study on biological characteristics of encephadynorokine

Objective The protective effects of eneephadynorokine on neuronal cell culture of ralembryonic brain were studied. Methods The cerebral cells of Wistar rats aged 16 days were cultured in vitro and studied in the following respects. 1)Histomorphology: different dosage of eneephadynerokine (1, 10 and 100 mg/L) were added in nerve cell culture. Then the cell morphology and ultramicrostrueture were studied under oplie and electron microscope respectively. 2)Estimation of nerve cell metabolic activity: the effect of different dosage of eneephadynorokine on metabolie activity of nerve cell was estimated by tetazolium(MTT) method. 3)Estimation of soluble protein level: effect of different dosage of eneephadynorokin on soluble protein level in nerve cell eytoplasm was measured by Bradford quantitative method for protein.4) Test ol enceph-dynorokine for anti-nerve cell apoptosis: the effects of eneephadynorokine and Austria cerebrolysin on the neuron specific enolase (NSE) expression were measured by NSE, qnantilative test. Results 1)It was showed in morphology that the eneephadynorokine could promote the proliferation and differentiation ol nerve cells, the number and lengtb of nerve cell processes were also inereased. 2)The effects of different dosage of eneephadynorokine on nerve cell growth, development and differentiation were showed that MTT metabolic rate and cytoplasmic protein level were increased. The dosage dependent correlation was showed obviously as increasing of en-eepphadynorokine dosage. The NSE expression was promoted by eneephadynorokine and helpful tor nerve cell differentiation. 3) The apoptosis resulted from antihypoxia and glulamie toxiration of nerve cell could be enhanced by eneephadynorokine. The effect of eneephadynorokine was superior than Austria cerobrolysin. Conclusion The anti-apoptosis effect of eneephadynorokine on nerve cells is superior than that of eere-brolysin in the similar experimental condition.