糖基化终末产物对人神经母细胞瘤细胞APP表达及Aβ生成的影响  

The effect of advanced glycation endproducts on the expression of amyloid precursor protein and β-amyloid protein in human neuroblastoma cells

作　　者：徐松[1] 高顺宗[2] 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内科,济南硕士研究生250021 [2]山东大学附属省立医院老年心内科,济南250021

出　　处：《卒中与神经疾病》2011年第2期67-71,102,共6页Stroke and Nervous Diseases

基　　金：国家自然科学基金(基金编号为30971036);山东省自然科学基金(基金编号为Y2008C13)

摘　　要：目的通过研究糖基化终末产物(AGEs-BSA)以及阻断其与特异性受体RAGE的结合对培养的人神经母细胞瘤细胞(SH-SY5Y)淀粉样前体蛋白(amyloid precursor protein,APP)的表达以及β淀粉样蛋白(β-amyloid,Aβ)生成的影响。方法以培养的SH-SY5Y细胞为模型,将细胞随机分为4组,空白对照组、BSA组、AGEs-BSA组、AGEs-BSA+抗RAGE中和抗体组。用MTT法观察细胞形态以及确定AGEs-BSA的最佳干预时间及浓度。用免疫细胞化学方法及免疫印迹方法来检测各组细胞内APP、RAGE表达和Aβ生成情况。结果不同蛋白浓度的BSA处理细胞24、48、72h,与空白对照组比较细胞MTT代谢率,APP、RAGE及Aβ的表达水平没有明显差异,(P>0.05);不同蛋白浓度的AGEs-BSA(>50μg/ml)处理细胞与BSA组比较,细胞MTT代谢率明显降低,并随蛋白浓度升高差异越明显,APP、RAGE、Aβ的表达水平较BSA组明显增加(P<0.05),预先用抗RAGE中和抗体(1∶100)1h后再加入AGEs-BSA,APP、Aβ的表达水平较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、RAGE、Aβ的表达和生成增加。通过阻断其与特异性受体RAGE的结合可以部分减少APP、Aβ的表达和生成。Objective To investigate the effect of advanced glycation endproducts （AGEs） and blocking AGEs-BSA and its acceptor（RAGE）＇s union on the expression of arnyloid precursor protein （APP） and 13-amyloid protein （Aβ） in cultred SH-SY5Y cell. Methods Raise the SH-SYSY cells as models and divide them into four groups randomly, the blank control group, the BSA group, the AGEs-BSA group, and AGEs-BSA ＋ th, anti-RAGE antibody group. To determine AGEs-BSA with the MTT metabolic rate form the best interventior time and the density. To examine the protein expression of APP,RAGE and Aβ in SH-SYSY cells by immunocytochemistry and Western blotting. Results Comparing with the blank group, treatment with concentration, of BSA for 24,48,72hours, there were no difference in MTT metabolic rate and the expression of APP, RAGE and Aβ protein（P〉0. 05）; Comparing with the BSA group, treatment with concentrations of AGEs-BSA （〉50/μg/ml） ,MTT metabolic rate reduced significantly in concentration-independent manner, and the expressior of APP,RAGE and Aβ protein increased significantly （P〈0. 05）. Comparing with the AGEs-BSA group, the cells exposed to anti-RAGE antibody（ 1 ： 100）for one hour before treated with AGEs-BSA, the expression ot APP and Aβ protein reduced significantly （P〈0. 05）. But comparing with the BSA group, the expression ot APP and Al3 protein increased significantly in the AGEs-BSA ＋ the anti-RAGE antibody group （P〈0. 05）.Conclusions AGEs-BSA could promote the production of APP, RAGE and Aβ protein, blocking AGEs-BSA and its acceptor（RAGE）＇s union could reduce the producttion of APP and Aβ protein.

关 键 词：阿尔茨海默病 糖基化终末产物 淀粉样前体蛋白 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]