川芎嗪显著改善阿尔茨海默病转基因细胞AD样病变及蛋白表达谱失调  

TMP significantly improves AD-like pathologies and proteome profiling dysregulation in a cell model of Alzheimer’disease

作　　者：高旭芳 许康 Xufang GAO;Kang XU(Department of Neurology,General Hospital of the Yangtze River Shipping and Wuhan Brain Hospital,No.5 Huiji Road,Wuhan,Hubei 430010,China)

机构地区：[1]长江航运总医院,武汉脑科医院神经内科,武汉430010

出　　处：《阿尔茨海默病及相关病杂志》2023年第4期264-271,共8页Chinese Journal of Alzheimer's Disease and Related Disorders

摘　　要：2,3,5,6-四甲基吡嗪(TMP)是一种广泛存在于传统中草药川芎(Liguisticum Wallichii)中的纯天然小分子化合物。虽然大量动物实验证据表明TMP可有效改善阿尔茨海默病(AD)样病理,但其具体机制尚不明确。目的:本研究旨在围绕线粒体相关蛋白质网络,探究TMP改善AD相关病理的分子机制。方法:将野生型(WT)和转染人源“瑞典”突变淀粉样蛋白前体蛋白(APPswe,以下简称APP)的小鼠神经瘤母细胞系和进行细胞培养,分为空白对照组(N2a/WT)、实验对照组(N2a/APP)和实验组(N2a/APP TMP),其中实验组设置0.2μM、2μM和20μM三个TMP给药浓度。溶剂或TMP处理24h后,提取细胞总蛋白进行Western blot检测和TMT标记蛋白质组学检测。结果:Western Blot结果显示,TMP能显著抑制N2a-APP细胞中人源“瑞典”突变淀粉样蛋白前体蛋白的表达。蛋白质组学分析表明,与野生型N2a/WT细胞相比N2a/APP细胞中共有65种差异表达蛋白(DEPs),其中33种蛋白质表达发生下调,32种蛋白质表达发生上调。生物信息学分析表明,差异表达蛋白参与多种AD病程相关生物学途径,包括线粒体功能、细胞骨架结构、ATP和GTP结合以及细胞凋亡。进一步Western Blot检测显示TMP上调N2a-APP细胞中氧化呼吸传递链复合物I、IV的表达水平,并表现出恢复复合物II、III和V组分表达水平的趋势。结论:TMP处理可以显著减少APP表达,同时可能通过调控线粒体蛋白质网络改善AD样症状,发挥神经元保护作用。2,3,5,6-tetramethylpyrazine(TMP)is a natural compound prevalent in the traditional Chinese herbal medicine,Liguisticum Wallichii.Although there is solid evidence that TMP has beneficial effect in treating animal model of AD,the mechanism behind remains elusive.Objective:Our research aims to study the molecular mechanism of AD-treating effect of TMP by focusing on mitochondria proteomic profile.Methods:We performed cell culture with N2a cell carrying human“Swedish”mutant amyloid precursor protein(N2a/APP),a solid cell model of AD that excessively express pathologicalβ-amyloid,and wild-type N2a cell.Cell models are grouped as white-control group(N2a/WT),experimental-control group(N2a/APP),and dosage group(N2a/APP TMP)in which three different dosages of TMP are set.After 24 hours of TMP or saline treatment,the total protein from each group of N2a cell are extracted for Western blot examination or TMT protein labeling.Results:Western Blot results show TMP can significantly inhibit the expression of APP.Results of proteomic analysis indicated total 65 differentially expressed proteins(DEPs)in N2a/APP cells compared to wild-type N2a/WT cells,including 33 decreased and 32 increased proteins.Bioinformatic analysis indicated that differentially expressed proteins are participating in several pathways including mitochondrial function,cytoskeleton structure,ATP and GTP binding,and apoptosis.We further examined the expression of specific proteins involved in mitochondrial electron respiratory chain by western blotting.As results,TMP treatment can significantly increase the expression level of complex I,IV and exhibited a tendency of restoring expression level of complex II,III and V component.Conclusion:TMP can significantly inhibit expression of APP and may have neuroprotective effect in modifying proteomic expression profile against AD pathology through different cellular mechanism,remarkably through regulating the mitochondrial functions.

关 键 词：阿尔茨海默病 2 3 5 6-四甲基吡嗪 氧化呼吸链 线粒体功能 

分 类 号：R74[医药卫生—神经病学与精神病学]