高糖对神经系统的影响指导.docVIP

摘要　目的　通过观察APP17肽和高糖环境对人神经母细胞瘤株SY5Y生长的影响，从细胞水平证实此肽有促进神经细胞生长的作用，并初步观察其对高糖引起神经元毒性作用的保护效应。方法　将细胞计数、MTT代谢率及LDH漏出率作为观察指标。结果　APP17肽可使细胞数和MTT代谢率明显增加，高糖环境下细胞数和MTT代谢率明显降低，LDH漏出率升高，此时加入APP17肽仍可使MTT代谢率增加。结论　①APP17肽具有促进神经元生长的作用，且能增加细胞线粒体活性，故可能是一种细胞代谢促进剂。②在无胰岛素的情况下，高糖环境对神经元有损伤作用，而APP17肽对神经元有一定的保护作用。 Effect of APP17 peptide on the neurotoxicity of high glucose Wang Rong,Zhang Jingyan,Zhang Dai,et al 　　(Capital University of Medical Sciences,Xuan Wu Hospital,Beijing Aging Research Laboratory,Beijing 100053) 　　abstract　Objective　To observe the effect of APP17 peptide and a high glucose environment on the growth of human neuroblastoma cells,SY5Y,to prove that this peptide can enhance neuronal growth from a cellular perspective,and to make preliminary observations of its preventive effect on the neurotoxicity of high glucose.Methods　Cells number,MTT metabolic rate and LDH leakage rate were assessed.Results　APP17 peptide significantly increased cell number and MTT metabolic rate.Cell number and MTT metabolic rate were reduced,and LDH leakage was increased in a high glucose environment,but the addition of APP17 peptide under such conditions can still raise MTT metabolic rate.Conclusion　①APP17 peptide had neurotrophic effect and promoted mitochondrial activity,therefore it may be a metabolic enhancer.②In the absence of insulin a high glucose environment was neurotoxic,and APP17 peptide was neuroprotective. 　　Key words　APP17 peptide　High glucose environment 　　APP是β-淀粉样肽(β-Amyloid,Aβ或β/A4)的前体蛋白(amyloid β/A4 protein precusor,APP)［1］。APP695由695个氨基酸组成［2］，其中319～335肽段为APP17肽(APP 17-mer peptide)［3］。已有报道，大鼠脑内注射此肽段两周，即可引起细胞学及行为学方面的改变，并能提高动物的学习记忆能力［4］。本实验室近年来对APP17肽进行了免疫组织化学方面的研究，结果表明它能明显改善糖尿病脑病动物海马神经元的退变(未发表资料)。葡萄糖作为动物体内能量来源之一，参与细胞生长代谢过程。但在缺乏胰岛素的情况下，高于体内正常水平的葡萄糖对神经元是否存在损伤作用，却少有报道［5］。本研究目的在于通过观察APP17肽和高糖环境对人神经母细胞瘤株(human neuroblastoma cells)SY5Y生长的影响，从细胞水平证实此肽促进神经细胞生长的作用，并初步观察其对高糖引起神经元毒性作用的保护效应。 材料和方法 　　一、材料 　　1.SY5Y细胞株：由瑞典卡罗琳斯卡研究所Bengt Winblad教授和裴进京博士赠送。细胞培养条件为MEM(Gibco BRL,41500-034)培养基中加入10%胎牛血清