U18666A对星形胶质瘤U373细胞APP代谢及Aβ生成的影响  

Effect of U18666A on APP metabolism and Aβgeneration in U373 cells of astroglioma

作　　者：杨宏艳[1] 都晓辉[1] 包亚男[1] 韩云峰 田华[1] YANG Hongyan;DU Xiaohui;BAO Ya′nan;HAN Yunfeng;Tian Hua(Pharmacy School,Qiqihar Medical University,Heilongjiang Province,Qiqihar161006,China;Public Health School,Qiqihar Medical University,Heilongjiang Province,Qiqihar161006,China)

机构地区：[1]齐齐哈尔医学院药学院,黑龙江齐齐哈尔161006 [2]齐齐哈尔医学院公共卫生学院,黑龙江齐齐哈尔161006

出　　处：《中国医药导报》2020年第22期4-7,共4页China Medical Herald

基　　金：黑龙江省教育厅科学技术研究项目(2018-KYYWF-0101);国家自然科学基金资助项目(81803757)。

摘　　要：目的观察胆固醇转运抑制剂U18666A(UA)对星形胶质瘤U373细胞淀粉样前体蛋白(APP)代谢及β淀粉样蛋白(Aβ)生成的影响。方法0、1、3、5μg/mL UA作用U373细胞24 h,或5μg/mL UA与U373细胞共培养0、12、24、48 h,Western blot检测APP及其代谢产物α-CTF、β-CTF的表达,酶联免疫吸附试验检测细胞内Aβ1-40含量,荧光酶法检测β-分泌酶活性。结果3、5μg/mL UA组APP蛋白表达水平、β-CTF和α-CTF蛋白表达量、Aβ1-40含量高于0μg/mL UA组,差异均有统计学意义(P<0.05或P<0.01)。共培养24、48 h组APP表达水平、β-CTF和α-CTF蛋白表达量、Aβ1-40含量高于共培养0 h组,差异均有高度统计学意义(均P<0.01)。5μg/mL UA组β-分泌酶活性高于对照组,差异有高度统计学意义(P<0.01)。结论UA通过增加β-分泌酶活性增强APP的表达和代谢,促进Aβ1-40的生成,加速阿尔茨海默病进程。Objective To investigate the effect of cholesterol transport inhibitor U18666A(UA)on amyloid precursor protein(APP)metabolism andβ-amyloid protein(Aβ)generation in U373 cells of astroglioma.Methods U373 cells were treated with UA at the dose of zero,one,three and fiveμg/mL for 24 h or 5μg/mL UA for different times(0,12,24,48 h).The expressions of APP and its metabolitesα-CTF andβ-CTF were detected using Western blot.The cellular levels of Aβ1-40 were measured with enzyme linked immunosorbent assay and the activity ofβ-secretase was determined by fluorescence assay.Results The expression levels of APP,α-CTF andβ-CTF,and Aβ1-40 in 5μg/mL UA group were higher than those in 0μg/mL UA group,and the differences were statistically significant(P<0.05 or P<0.01).The expression levels of APP,α-CTF andβ-CTF,and Aβ1-40 in co-cultured 24 h and 48 h groups were significantly higher than those in co-cultured 0 h group,and the differences were highly statistically significant(all P<0.01).The activity ofβ-secretase in 5μg/mL UA group was significantly higher than that in the control group(P<0.01).Conclusion UA enhances APP expression and metabolism by increasingβ-secretase activity,promotes Aβ1-40 generation,accelerates the pathological process of Alzheimer's disease.

关 键 词：胆固醇 U18666A U373细胞 淀粉样前体蛋白 Β淀粉样蛋白 

分 类 号：R749.16[医药卫生—神经病学与精神病学]