Aβ寡聚体调控M1胆碱受体下游信号转导通路的机制研究  

The Mechanism of AβOligomers Regulating the Downstream Signaling Transduction of M1 Muscarinic Acetylcholine Receptor

作　　者：蒋敏洁 张永芳 王冬雪 王昊 徐见容 赵兰雪 JIANG Min-jie;ZHANG Yong-fang;WANG Dong-xue;WANG Hao;XU Jian-rong;ZHAO Lan-xue(Department of Pharmacology and Chemical Biology,Shanghai Jiao Tong University School of Medicine,Shanghai,200025,China;Institute of Interdisciplinary Integrative Biomedical Research,Shanghai University of Traditional Chinese Medicine,Shanghai,201204,China;Academy of integrative medicine,Shanghai University of Traditional Chinese Medicine,Shanghai,201204,China)

机构地区：[1]上海交通大学医学院药理学与化学生物学系,上海200025 [2]上海中医药大学交叉科学研究院,上海201204 [3]上海中医药大学中西医结合研究院,上海201204

出　　处：《现代生物医学进展》2022年第17期3201-3207,共7页Progress in Modern Biomedicine

基　　金：上海市卫计委临床研究专项(20194Y0056);国家自然科学基金面上项目(82173798)。

摘　　要：目的:阿尔茨海默病(Alzheimer's disease,AD)病理状态下M1胆碱受体功能受损,无法对配体做出正确响应。本研究模拟AD早期Aβ寡聚体富集的病理状态,考察对M1胆碱受体下游信号通路关键分子活性的影响,以期为AD病理状态下M1胆碱受体信号偶联障碍机制提供新的见解。方法:制备Aβ寡聚体(Aβoligomers,AβOs),1μM AβOs预处理CHO-M1细胞30 min、24 h和48 h,加入胆碱受体激动剂乙酰胆碱或者卡巴胆碱激活M1胆碱受体,利用Ca^(2+)检测实验、IP-One-Gq KIT检测实验及Western blot实验检测受体激动后下游关键信号分子Ca^(2+)、IP_(1)的释放量以及ERK磷酸化的动态变化。结果:AβOs预处理导致激活细胞内M1胆碱受体其下游Ca^(2+)和IP_(1)的释放量减少、ERK被更快激活。结论:AβOs预处理导致激活M1胆碱受体下游关键信号分子释放水平、磷酸化水平异常,揭示在阿尔茨海默病早期M1胆碱受体便出现功能障碍,无法正确激活信号转导通路。Objective:M1 muscarinic receptors are functional impairment in Alzheimer's disease(AD)and fail to respond to ligands properly.This study simulated the pathological state of Aβoligomers enrichment in early AD,and investigated the effects on the activities of key molecules in the downstream signaling pathway of M1 muscarinic receptors,in order to provide new insights into the mechanism of M1 muscarinic receptors coupling disorder in AD.Methods:Aβoligomers were successfully prepared in vitro.CHO-M1 cells were pretreated with 1μM Aβoligomers for 30 min,24 h,and 48 h,and then ACh or CCh agonists were adopted to activate the M1 muscarinic receptors.Calcium mobilization assay,IP-One-Gq KIT and Western blot were utilized to detect the downstream key signaling molecules Ca^(2+),IP_(1)and the dynamic changes of ERK phosphorylation after receptors activation.Results:Aβoligomers preconditioning resulted in reduced Ca^(2+)and IP_(1)release and earlier activation of ERK in the downstream of the activated M1 muscarinic receptors.Conclusion:Aβoligomers pretreatment lead to abnormal release and phosphorylation levels of key signaling molecules downstream of activated M1 muscarinic receptors,revealing that M1 muscarinic receptors are dysfunctional in the early stages of AD and cannot correctly activate signaling transduction pathways.

关 键 词：AΒ寡聚体 M1毒蕈碱型乙酰胆碱受体 信号转导通路 阿尔茨海默病 G蛋白偶联受体 

分 类 号：R-33[医药卫生] R749.16[生物学—分子生物学] Q73R966