基于UPLC-Q-TOF-MS的补阳还五汤抗慢性炎症大鼠尿液代谢组学研究  

Metabolomic study on urine of chronic inflammation rats treated with Buyang Huanwu Decoction based on UPLC-Q-TOF-MS

作　　者：尤心怡 姜丽[1,2] 王文凤 徐霞 张寿文 刘红宁 严小军[1] 聂鹏[1] 李冰涛[1,2] 徐国良[1,2] YOU Xin-yi;JIANG Li;WANG Wen-feng;XU Xia;ZHANG Shou-wen;LIU Hong-ning;YAN Xiao-jun;NIE Peng;LI Bing-tao;XU Guo-liang(Jiangxi Provincial Key Laboratory of Traditional Chinese Medicine Etiopathogenisis,Research Center for Differentiation and Development of Traditional Chinese Medicine Basic Theory,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Key Laboratory of Pharmacology of Traditional Chinese Medicine in Jiangxi,Nanchang 330004,China;Qilu Hospital of Shandong University Dezhou Hospital,Dezhou 253000,China;Research Center for Traditional Chinese Medicine Resources and Ethnic Minority Medicine,Jiangxi University of Chinese Medicine,Nanchang 330004,China)

机构地区：[1]江西中医药大学中医基础理论分化发展研究中心江西省中医病因生物学重点实验室,江西南昌330004 [2]江西省中药药理学重点实验室,江西南昌330004 [3]山东大学齐鲁医院德州医院,山东德州253000 [4]江西中医药大学中药资源与民族药研究中心,江西南昌330004

出　　处：《中国中药杂志》2023年第19期5345-5355,共11页China Journal of Chinese Materia Medica

基　　金：江西省卫生科技计划项目(SKJP220210795);江西省教育厅科学技术研究项目(GJJ2200956);江西中医药大学校级科技创新团队发展计划项目(CXTD22007);江西省第四批中青年中医药骨干人才计划项目。

摘　　要：基于超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)探讨补阳还五汤(Buyang Huanwu Decoction,BYHWD)对大肠杆菌脂多糖(LPS)诱导的慢性炎症大鼠尿液中内源性标志物的影响,旨在从代谢组学的角度阐述BYHWD治疗慢性炎症的分子作用机制。雄性SD大鼠随机分成正常组、模型组以及BYHWD低、中、高剂量组(7.5、15、30 g·kg^(-1))。模型组及各药物组每周首日尾静脉注射LPS 200μg·kg^(-1),并于次日灌胃BYHWD,每日1次,连续4周。给药结束收集尿液样品,应用UPLC-Q-TOF-MS分析各组大鼠的尿液代谢轮廓,结合主成分分析(PCA)、偏最小二乘法判别分析(PLS-DA)和正交偏最小二乘法判别分析(OPLS-DA)等多元统计分析方法,分析BYHWD对内源性代谢物的影响;结合单因素方差分析(One-way ANOVA)和变量投影重要度(VIP)筛选出与慢性炎症相关的潜在生物标志物;通过MetaboAnalyst 5.0对鉴定出来的生物标志物进行通路及富集分析。结果显示,在大鼠尿液中筛选和鉴定出25个潜在的生物标志物,与正常组相比,模型组中有14个物质的含量显著上升(P<0.05),11个物质的含量显著下降(P<0.05);BYHWD能够使绝大部分内源性生物标志物呈现良好回调趋势。与模型组比较,BYHWD可显著下调其中的13个生物标志物(P<0.05),上调其中的10个生物标志物(P<0.05)。代谢产物主要与泛酸和辅酶A生物合成、色氨酸代谢、视黄醇代谢、丙酸代谢等代谢通路有关。BYHWD对LPS诱导的慢性炎症具有治疗作用,可能与改善内源性代谢物的水平,提高机体抗炎、抗氧化能力,恢复机体正常代谢活动有关。The study investigated the effect of Buyang Huanwu Decoction(BYHWD) on endogenous biomarkers in the urine of rats with chronic inflammation induced by lipopolysaccharide(LPS) using ultra-high performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry(UPLC-Q-TOF-MS),aiming to elucidate the molecular mechanism underlying the therapeutic effect of BYHWD on chronic inflammation from a metabolomics perspective.Male SD rats were randomly divided into a normal group,a model group,and low-,medium-,and high-dose BYHWD groups(7.5,15,and 30 g·kg^(-1)).The model group and BYHWD groups received tail intravenous injection of LPS(200 μg·kg^(-1)) on the first day of each week,followed by oral administration of BYHWD once a day for four consecutive weeks.Urine samples were collected at the end of the administration period,and UPLC-Q-TOF-MS was used to analyze the metabolic profiles of the rat urine in each group.Multivariate statistical analysis methods such as principal component analysis(PCA),partial least squares-discriminant analysis(PLS-DA),and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to analyze the effect of BYHWD on endogenous metabolites.One-way ANOVA and variable importance for the projection(VIP) were used to screen for potential biomarkers related to chronic inflammation.The identified biomarkers were subjected to pathway and enrichment analysis using MetaboAnalyst 5.0.A total of 25 potential biomarkers were screened and identified in the rat urine in this experiment.Compared with the normal group,the model group showed significant increases in the levels of 14 substances(P<0.05) and significant decreases in the levels of 11 substances(P<0.05).BYHWD was able to effectively reverse the trend of most endogenous biomarkers.Compared with the model group,BYHWD significantly down-regulated 13 biomarkers(P<0.05) and up-regulated 10 biomarkers(P<0.05).The metabolic products were mainly related to the biosynthesis of pantothenic acid and coenzyme A,tryptophan metabolism,ret

关 键 词：补阳还五汤 慢性炎症 代谢组学 UPLC-Q-TOF-MS 代谢通路 代谢机制 

分 类 号：R285.5[医药卫生—中药学]