糖基化终末产物促SH-SY5Y细胞β-淀粉样蛋白生成及相关机制  

Effect of advanced glycation end products on expression of the β-amyloid protein in SH-SY5Y cells and its related mechanism

作　　者：徐松[1] 高顺宗[1] 刘雪平[1] 王美霞[1] 董传芳[1] 侯亮[1] 袁树华[1] 

机构地区：[1]山东大学附属省立医院老年神经科,济南250021

出　　处：《山东大学学报（医学版）》2011年第6期33-37,45,共6页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助项目(30971036);山东省自然科学基金资助项目(Y2008C13)

摘　　要：目的通过研究糖基化终末产物(AGEs-BSA)对培养的人神经母细胞瘤细胞(SH-SY5Y细胞)β-淀粉样蛋白(Aβ)的生成,以及淀粉样前体蛋白(APP)及相关酶—β-分泌酶(BACE1)、γ-分泌酶(PS1)的表达的影响,在体外水平探讨AGEs-BSA在阿尔茨海默病(AD)发病中的作用及其可能的机制。方法以培养的SH-SY5Y细胞为模型,将细胞随机分为4组。用MTT实验得到的AGEs-BSA最佳干预时间及浓度干预细胞,用免疫细胞化学方法及ELISA方法观察及检测各组细胞内Aβ1-40、Aβ1-42表达,用免疫印迹法检测各组细胞内APP、BACE1、PS1变化。结果 BSA组与空白对照组相比APP、BACE1、PS1、Aβ的表达无明显差异(P>0.05);AGEs-BSA组与BSA组相比APP、BACE1、PS1、Aβ的表达明显增加(P<0.05);AGEs-BSA+抗RAGE中和抗体组APP、BACE1、PS1、Aβ的表达较单纯AGEs-BSA组明显减少(P<0.05),但仍高于BSA组(P<0.05)。结论 糖基化终末产物能够促使SH-SY5Y细胞中APP的表达增加,并通过上调BACE1、PS1的活性使Aβ生成增加。通过阻断其与特异性受体RAGE的结合可以部分减少APP、BACE1、PS1及Aβ的表达和生成。Objective To investigate the effect of advanced glycation end products （AGEs） on expressions of the β- amyloid protein （Aβ） and its related enzymes in cultured SH-SY5Y cells, and explore the effect and possible mecha- nism of AGEs on Alzheimer＇s disease（AD） the cell level. Methods Cultured SH-SY5Y ceils were randomly divided into four groups： the blank control group, the AGE-modified bovine serum albumin （AGEs-BSA） group, the AGEs- BSA ＋ anti-receptor for advanced glycation end products（RAGE） group and the BSA group. The MTT metabolic rate was employed to determine cells＇ growth and best concentration and time of the AGEs-BSA. Immunocytochemistry and ELISA were used to observe expressions of Aβ1-40 and Aβ1-42. Western blot was employed to examine changes of the amyloid precursor protein （ APP）, β- secretion enzymel （ BACE1 ） and presenilinl （ PSI ） in SH-SY5Y cells. Results There was no difference in APP, BACE1, PS-land Aβ between the blank control group and the BSA group（P 〉 0.05）. Immunocytochemistry and ELISA results indicated that expression of A13 in cells was significantly higher in AGEs-BSA and AGEs-BSA ＋ antiRAGE groups than in the BSA group （ P 〈 0.05 ）, and it was lower in the AGEs-BSA ＋ antiRAGE group than that in the AGEs-BSA group （ P 〈 0.05 ）. Western blot showed that APP, BACE1 and PSI levels in SH-SY5Y cells were elevated in AGEs-BSA and AGEs-BSA ＋ antiRAGE groups compared with the BSAgroup（ P 〈0.05 ）, and concentrations of them in the AGEs-BSA ＋ antiRAGE group were lower than those in the AGEs- BSA group（P 〈 0.05）. Conclusion AGEs-BSA promotes expression of APP, and it promotes expression of A[~ by up-regulating activities of BACE1 and PS1. The blocking combination of AGEs-BSA and its receptor（RAGE） reduces expressions of APP, BACE1, PSI and Aβ.

关 键 词：阿尔茨海默病 糖基化终末产物 Β-淀粉样蛋白 

分 类 号：R592[医药卫生—老年医学] R742[医药卫生—内科学]