基于非靶向代谢组学检测探讨肾精亏虚型颈动脉粥样硬化患者的代谢特征
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摘要
目的:筛选肾精亏虚型颈动脉粥样硬化患者与健康人群之间的差异代谢物,为其诊断及发病机制研究提供科学依据。方法:纳入肾精亏虚型颈动脉粥样硬化患者及健康志愿者各8例,采集所有受试者的血浆样本。应用超高压液相色谱-电喷雾-四级杆飞行时间串联质谱分析法(UHPLC-Q-TOF MS)对血浆样本进行全谱分析,采用XCMS软件对数据进行峰提取和代谢物鉴定;运用主成分分析、判别分析等统计方法筛选差异代谢物,再将差异代谢物提交到KEGG网站,进行相关通路分析。结果:正、负离子模式下,肾精亏虚型颈动脉粥样硬化患者与健康志愿者间具有显著性差异的代谢物共17种:精胺、焦谷氨酸、肉碱、精氨酸、肌酐、肌酸、三甲基甘氨酸、牛磺鹅胆酸盐、泛酸酯、硫酸犬尿素、左旋瓜氨酸、甘氨胆酸、γ-谷酰基-L-苯基丙氨酸、雌素酮-3-硫酸盐、D-阿洛糖、柠檬酸盐、乙酰-DL-亮氨酸;所有差异性代谢物共参与78条代谢通路,其中3种及以上差异性代谢物同时参与的代谢通路共19条。结论:肾精亏虚型颈动脉粥样硬化患者较健康志愿者的代谢物谱发生了显著性改变,这些差异代谢物及其所涉及的代谢通路可能成为未来证型和药物靶点研究的方向。
Objective: To screen the differential metabolites between patients of carotid atherosclerosis with syndrome of kidney essence deficiency and healthy volunteers,and provide scientific basis for its diagnosis and pathogenesis. Methods: Eight patients of carotid atherosclerosis with syndrome of kidney essence deficiency and 8 volunteers were included. The plasma samples of all subjects were collected. UHPLC-Q-TOF MS was carried out to analyze the full spectrum of the samples,and XCMS software was used to extract the peak and identify the metabolites. Principal component analysis,discriminant analysis and other statistical methods were used to screen the differential metabolites. The differential metabolites were submitted to KEGG website for relevant pathway analysis. Results: In the positive and negative ion mode,there were 17 kinds of significant differential metabolites between the patients of carotid atherosclerosis with syndrome of kidney essence deficiency and the healthy volunteers,including spermine,L-pyroglutamic acid,L-carnitine,L-arginine,creatinine,creatine,trimethylglyine,taurochenodeoxycholate,pantothenate,L-kynurenine,L-citrulline,glycocholic acid,γ-L-glutamyl-Lphenyal anine,estrone-3-sulfate,D-allose,citrate,acetyl-DL-leucine. All of the significant differential metabolites were involved in the 78 metabolic pathways,and the number of the metabolic pathways co-involved by three and more than three differential metabolites was 19.Conclusion: There are significant differences on the metabolite spectrum between the patients of carotid atherosclerosis with syndrome of kidney essence deficiency and the healthy volunteers,and these differential metabolites and the relevant metabolic pathway may become the research direction of syndrome and drug target in the future.
Objective: To screen the differential metabolites between patients of carotid atherosclerosis with syndrome of kidney essence deficiency and healthy volunteers,and provide scientific basis for its diagnosis and pathogenesis. Methods: Eight patients of carotid atherosclerosis with syndrome of kidney essence deficiency and 8 volunteers were included. The plasma samples of all subjects were collected. UHPLC-Q-TOF MS was carried out to analyze the full spectrum of the samples,and XCMS software was used to extract the peak and identify the metabolites. Principal component analysis,discriminant analysis and other statistical methods were used to screen the differential metabolites. The differential metabolites were submitted to KEGG website for relevant pathway analysis. Results: In the positive and negative ion mode,there were 17 kinds of significant differential metabolites between the patients of carotid atherosclerosis with syndrome of kidney essence deficiency and the healthy volunteers,including spermine,L-pyroglutamic acid,L-carnitine,L-arginine,creatinine,creatine,trimethylglyine,taurochenodeoxycholate,pantothenate,L-kynurenine,L-citrulline,glycocholic acid,γ-L-glutamyl-Lphenyal anine,estrone-3-sulfate,D-allose,citrate,acetyl-DL-leucine. All of the significant differential metabolites were involved in the 78 metabolic pathways,and the number of the metabolic pathways co-involved by three and more than three differential metabolites was 19.Conclusion: There are significant differences on the metabolite spectrum between the patients of carotid atherosclerosis with syndrome of kidney essence deficiency and the healthy volunteers,and these differential metabolites and the relevant metabolic pathway may become the research direction of syndrome and drug target in the future.
引文
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[2]宫兵,吴东垣,王丽岩,等.颈动脉粥样硬化与冠心病严重程度的相关性[J].中国实验诊断学,2014,18(4):601-603.
[3]施迎兵.颈动脉粥样硬化斑块与脑梗死相关性分析[J].中国实用医药,2014,9(7):92-93.
[4]吴凤英,胡静.脑梗死与颈动脉粥样硬化斑块及其相关因素分析[J].中国实用神经疾病杂志,2010,13(1):26-28.
[5]陈明荃,顾耘,徐辉.颈动脉粥样硬化患者中医证候分布探讨[J].中华中医药学刊,2012,30(6):1394-1396.
[6]杜文婷,顾耘,徐辉,等.基于多元统计学方法对颈动脉粥样硬化中医证候的初步研究[J].辽宁中医杂志,2014,41(9):1859-1861.
[7]中国医师协会超声医师分会.血管超声检查指南[J].中华超声影像学杂志,2009,18(10):911-920.
[8]郑筱萸.中药新药临床研究指导原则(试行)[M].北京:中国医药科技出版社,2002:77-85.
[9]POULTER N.Global risk of cardiovascular disease[J].Heart,2003,89(supple2):ii2-ii5.
[10]薛巧云,殷春萍.颈动脉粥样硬化易损斑块的中医证型分布规律探讨[J].中国医药指南,2015,13(5):215-216.
[11]江春迎,杨康敏,杨柳,等.金黄地鼠动脉粥样硬化发展过程中的NMR代谢组学研究[J].药学学报,2013,48(4):495-502.
[12]VALLEJO M,GARCIA A,TUNON J,et al.Plasma fingerprinting with GC-MS in acute coronary syndrome[J].Anal Bioanal Chem,2009,394(6):1517-1524.
[13]MIAO H,CHEN H,ZHANG X,et al.Urinary metabolomics on the biochemical profiles in diet-induced hyperlipidemia rat using ultraperformance liquid chromatography coupled with quadrupole time-of-flight SYNAPT high-definition mass spectrometry[J].J Anal Methods Chem,2014,2014:184162.