基于代谢组学和质谱成像法考察烟碱暴露对小鼠脑内内源性代谢物的影响  

Effects of nicotine exposure on endogenous metabolites in mouse brain based on metabolomics and mass spectrometry imaging

作　　者：郭露露 张辰 黄延俊 刘兴余 刘德水 龙腾 孙晋浩 刘绍锋 李中皓 王加忠 毛健 GUO Lulu;ZHANG Chen;HUANG Yanjun;LIU Xingyu;LIU Deshui;LONG Teng;SUN Jinhao;LIU Shaofeng;LI Zhonghao;WANG Jiazhong;MAO Jian(Beijing Life Science Academy Co.,Ltd.,Beijing 102209,China;Department of Nutrition and Health,China Agricultural University,Beijing 100193,China;Technology Center,China Tobacco Guizhou Industrial Co.,Ltd.,Guiyang 550009,China)

机构地区：[1]北京生命科技研究院有限公司,北京102209 [2]中国农业大学营养与健康系,北京100193 [3]贵州中烟工业有限责任公司技术中心,贵州贵阳550009

出　　处：《色谱》2025年第4期363-371,共9页Chinese Journal of Chromatography

基　　金：国家自然科学基金(32072344,32272455);中国烟草总公司科技重点研发项目(110202102001);国家烟草专卖局创新平台科研活动稳定支持专项(国烟科[2021]171号).

摘　　要：烟碱具有明显的神经药理学活性,对多种中枢神经系统疾病具有干预作用,然而烟碱入脑后对脑区内源性代谢物的影响尚不清楚。研究烟碱暴露后对小鼠脑内内源性代谢物的影响,可为阐明烟碱的多重生理学效应提供更加全面的物质基础。本文基于超高效液相色谱-串联质谱(UHPLC-MS/MS)及代谢组学分析方法,系统考察了烟碱多次暴露后对小鼠脑内内源性代谢产物的影响,并进一步采用质谱成像法,直观获取烟碱引发的差异性代谢物在不同脑区的分布及变化差异。实验结果如下:本研究共鉴定到759种代谢物,烟碱暴露后共筛选到575个显著差异性代谢物,其中434个代谢物下调,141个代谢物上调;通过京都基因和基因组数据库(Kyoto Encyclopedia of Genes and Genomes,KEGG)通路富集分析表明烟碱暴露后主要影响到脑内必需氨基酸代谢、脂质代谢、核苷酸代谢、碳水化合物代谢、辅因子和维生素代谢以及其他氨基酸代谢通路;进一步通过质谱成像法分析差异性代谢物在脑内的分布特征,结果表明烟碱暴露后全脑范围内胆碱、丝氨酸、天冬氨酸及苹果酸含量都被显著下调,皮层区域牛磺酸、海马区域乙酰基肉碱以及纹状体区域的腺苷类物质均被显著影响。这些结果为进一步揭示烟碱暴露对中枢神经系统的影响提供了新的实验依据。Nicotine,the principal alkaloid in tobacco,exhibits significant central nervous system activity and induces a wide array of physiological effects.In addition to its well-documented role in tobacco dependence,previous studies have suggested that nicotine also has diverse pharmacological properties.These include alleviating symptoms associated with Parkinson’s disease,potentially reducing the risk of Alzheimer’s disease,mitigating oxidative stress,as well as anti-inflammatory and anxiolytic effects.Neuroscientists frequently use an array of molecular biology techniques to elucidate the mechanisms responsible for the effects of nicotine on the central nervous system.However,disease onset is invariably accompanied by metabolic dysfunction,and organisms often exhibit complex and unpredictable responses to pharmacological stimuli.As a bioactive alkaloid with potent pharmacological properties,nicotine is able to cross the blood-brain barrier and induce brain-compound changes,which serves as the basis for its effects on the central nervous system.Consequently,examining the extensive impact of nicotine exposure on endogenous metabolites and metabolic pathways in the brain is an indispensable step toward providing a more robust foundation for understanding the complex physiological effects of nicotine.In this study,an ultra-high performance liquid chromatography-tandem mass spectrometry(UHPLC-MS/MS)metabolomic-analysis method was established to systematically examine the effects of repeated nicotine exposure on endogenous metabolites in mouse brains.Two chromatographic systems fitted with Acquity UPLC BEH HILIC(150 mm×2.1 mm,1.7μm)and BEH C18(150 mm×2.1 mm,1.7μm)columns were used to determine the nicotine present in samples.As a result,the established UHPLC-MS/MS method identified a total of 759 endogenous metabolites.Compared with the saline group,nicotine exposure resulted in 575 significantly different metabolites,with 434 metabolites down-regulated and 141 up-regulated.Further pathway-enrichment analysis using

关 键 词：超高效液相色谱-串联质谱 代谢组学 质谱成像 烟碱 鼠脑 

分 类 号：O658[理学—分析化学]