刺五加皂苷对淀粉样前体蛋白APP酶解通路的影响  

Effect of Acanthopannx Senticousus Saponins on processing Pathway of Amyloid precursor protein

作　　者：刘芳[1] 李净洋 邱烨[1] 许妍姬[1] 

机构地区：[1]延边大学医学院预防医学教研室,吉林延吉133002

出　　处：《中国实验诊断学》2017年第12期2173-2176,共4页Chinese Journal of Laboratory Diagnosis

基　　金：吉林省教育厅科学技术研究项目(吉教科合字2016-279号)

摘　　要：目的研究刺五加皂苷对APP(Amyloid Precursor Protein,APP)酶解通路的影响。方法 (2015.9-2016.5)用12.5mg/L、25mg/L、50mg/L刺五加皂苷处理胚鼠皮层神经元细胞,采用酶联免疫吸附测定法(enzyme linked immunosorbent assay,ELISA)检测γ-分泌酶活性;另用脂质体2000转染构建APP695高表达CHO细胞系,用G418(500mg/L)进行筛选获得APP695-CHO稳转染细胞,用12.5 mg/L、25 mg/L、50 mg/L刺五加皂苷处理稳转染细胞,48h后,ELISA法检测细胞培养上清中Aβ1-40含量,用Western-blot法检测细胞裂解液中APP-N端片段蛋白的表达。结果在胚鼠皮层神经元细胞与对照组相比,25 mg/L、50 mg/L的刺五加皂苷能够抑制内源性γ-内切酶的活性,而且随着浓度的升高,抑制作用增强;在APP695-CHO稳转染细胞中,Aβ1-40的分泌量增高,APP-N端片段的蛋白表达水平减少,不同剂量刺五加皂苷能缓解这些变化。结论刺五加皂苷能抑制内源性γ-内切酶减少Aβ1-40的生成,对APP酶解起到一定的影响,对阿尔茨海默病的发病及进展有一定的保护作用。Objective To study the effects of Acanthopannx Senticousus Saponins（ASS）on processing pathway of amyloid precursor protein（APP）.Methods （2015.9-2016.5）12.5 mg/L,25 mg/L,50 mg/L of ASS were treated to primary culturedrat cortical neuron cells,the activity of γ-secretase was measured by ELISA analysis;The APP695 gene was transfected into CHO cells using Lipper 2000 reagent.Stably transfected cells were sclected by G418（500 mg/L）.12.5 mg/L,25 mg/L,50 mg/L of ASS were treated to stably transfected cellsfor 48 hours,ELISA assay was used to detected Aβ1-40 concentration in the supernatant of cell culture medium.Western-blot analysis were used to detected the APP-N terminal fragment protein expression from cell lysate.Results In the primary cultured cortical neuron cells, 25 mg/L,50 mg/L ASS inhibited the activity of γ-secretase compared with control group,and,the effect of inhibition were enhanced as the concentration increased.The concentration of Aβ1-40 was incresedand the expression of APP-N ter-minal fragment protein were decreased in APP695-CHO stably transfected cells,which all changes were alleviated by different dose of ASS.Conclusion ASS could inhibit the activity of endogenousγ-secretase to decreasing Aβ1-40 produc-tion and influence APP processing may have an protective effect on Alzheimer＇s disease pathology.

关 键 词：刺五加皂苷 Β淀粉样蛋白 γ-内切酶 阿尔茨海默病 APP-N端片段 

分 类 号：R285.5[医药卫生—中药学]