短肽N328-332对人神经母细胞瘤株SY5Y生长的影响  

Effect of amyloid beta-protein precursor N end 328-332 on the growth of human neuroblastoma cells SY5Y

作　　者：姚洁[1] 张景艳[2] 姬志娟[2] 王蓉[2] 盛树力[2] 

机构地区：[1]首都医科大学宣武医院检验科,北京市100053 [2]首都医科大学宣武医院北京脑老化重点实验室,北京市100053

出　　处：《中国临床康复》2006年第26期89-91,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家科技部"九七三"项目(2000057010)~~

摘　　要：目的:分析淀粉样β蛋白前体蛋白N端328-332对人神经母细胞瘤株SY5Y生长的影响,在体外寻找具有神经营养作用、促进神经细胞生长的最适浓度,为进一步细胞损伤的保护及药物体内研究提供理论依据。方法:细胞分组:根据实验目的不同,分组处理如下:第一步分为对照组、短肽N328-3322.5,5,10,20,40,80,160μmol/L组。第二步分为对照组、短肽N328-33240μmol/L组。以MTT代谢率测定细胞存活率以筛选肽对神经细胞生长促进的最佳有效浓度。以MTT代谢率测定细胞存活率、细胞计数观察细胞增长情况、乳酸脱氢酶漏出率观察细胞死亡率和WesternBlotting检测P-CREB、Bcl-2表达水平为观察指标,分析短肽N328-332对人神经母细胞瘤株SY5Y生长的影响。结果:①从10μmol/L起,短肽N328-332即对SY5Y细胞生长有促进作用,其最小有效浓度为20μmol/L,40μmol/L促进作用最强,80μmol/L开始作用减弱,故选用40μmol/L浓度作为以下试验浓度。②短肽N328-33240μmol/L加药组代表细胞存话率的A值高于对照组,组间比较差异有显著性意义(0.9741±0.0047,0.6001±0.0019,P<0.01。③短肽N328-33240μmmol/L加药组乳酸脱氢酶漏出率明显低于对照组,组间比较差异有显著性意义(84.9135±31.5759,199.6252±40.7273,P<0.01。④从接种第4天始,短肽N328-332组SY5Y细胞数增加,与对照组相比,差异有显著性意义(P<0.05)。⑤短肽N328-33240μmol/L组P-CREB、Bcl-2蛋白表达高于对照组(37424.08±83.59,34957.24±78.03,19110.24±45.80,2761.36±58.61,P<0.05)。结论:短肽N328-332对SY5Y细胞具有神经营养作用,40μmol/L浓度,效果优于其他浓度。AIM： To analyze the effects of amyloid beta-protein precursor N end 328- 332 on the growth of human neuroblastoma （HNB） SY5Y, and find in vitro the neurotrophic effects as well as the best concentration for growth of neurocytes, so as to provide the theoretical basis for the protection of cellular injury and in vivo research of medicament. METHODS： Grouping： cells were divided into groups according to the experimental purpose： In the first step, cells were divided into control group, 2.5, 5, 10, 20, 40, 80, 160 μmol/L APP 5-mer peptide group . In the second step, they were divided into control group and 40 μmol/L APP 5-mer peptide group. MMT metabolic rate was used to determine the cell survival rate so as to screen the best concentration for promoting the growth of neurocytes. MMT metabolic rate was adopted to determine the survival rate of cells and cell counting to detect the cell growth and lactate dehydrogenase （LDH） leakage rate, so as to observe the cell.death rate. The levels of P-CREB and Bcl-2 expression were inspected with Western Blotting and taken as the observation indicators. The effects of APP 5-mer peptide on HNB SY5Y growth. RESULTS： ①APP 5-mer peptide could promote the growth of SY5Y cells from 10 μmol/L, and the minimum concentration was 20 μmol/L, the effects of APP 5-mer peptide at 40 μmol/L was the best, and began to decrease at 80μmol/L. Therefore, the concentration below 40 μmol/L was taken as the test concentration. ②Compare with the normal control group, the A value of survival rate in administration group of 40 μmol/L APP 5mer peptide group was much higher, and differences among groups were significant （0.974 1±0.004 7,0.600 1±0.001 9,P 〈 0.01）.③The leaking rate of LDH in administration group of 40 μmol/L APP 5-mer peptide group was obviously lower than the control group, and there were remarkable differences among groups （84.913 5±31.575 9,199.625 2±40.727 3, P 〈 0.01）.④From the 4^th day of inoculation, the cell count of SY5Y cells in

关 键 词：淀粉样Β蛋白 神经母细胞瘤/病理学 细胞 培养的 

分 类 号：R749.4[医药卫生—神经病学与精神病学]