基于UPLC-Q-TOF/MS的黄连解毒汤抗动脉粥样硬化大鼠血浆代谢组学研究  

Metabolomics Study of Huanglian Jiedu Decoction on High Fat-induced Atherosclerosis Rats by UPLC-Q-TOF/MS

作　　者：姜丽[1,2,3] 李艳 李冰涛[1,2,3] 张启云[1,2] 余兰彬 陈雨[1] 王玮玮 熊艳玲 徐国良 JIANG Li;LI Yan;LI Bingtao;ZHANG Qiyun;YU Lanbin;CHEN Yu;WANG Weiwei;XIONG Yanling;XU Guoliang(Research Center for Differentiation and Development of TCM Basic Theory,Jiangxi University of Traditional Chinese Medicine,Nanchang 330004 Jiangxi,China;Jiangxi Provincial Key Laboratory of TCM Etiopathogenisis,Nanchang 330004 Jiangxi,China;Key Laboratory of Pharmacology of Traditional Chinese Medicine in Jiangxi,Nanchang 330004 Jiangxi,China;Department of Pharmacy,Zhejiang Hospital,Hangzhou 310013 Zhejiang,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,江西南昌330004 [2]江西省中医病因生物学重点实验室,江西南昌330004 [3]江西省中药药理学重点实验室,江西南昌330004 [4]浙江医院药剂科,浙江杭州310013

出　　处：《中药新药与临床药理》2020年第6期694-701,共8页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金青年项目(81703823);江西省自然科学基金项目(20192BAB205110);江西省卫生厅中医药科技计划项目(2018B131);江西省教育厅科学技术研究项目(GJJ170753);江西省中药学双一流学科项目(JXSYLXK-ZHYAO120)。

摘　　要：目的基于超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF/MS)技术探讨黄连解毒汤对动脉粥样硬化大鼠血浆中内源性标志物的影响,旨在寻找差异的潜在生物标志物群,以阐释黄连解毒汤抗动脉粥样硬化的分子机制。方法将雄性SD大鼠随机分成正常对照组、模型组及黄连解毒汤低、中、高剂量组(1.5、3、6 g·kg^-1)。采用高脂饲料+定期注射维生素D3复制动脉粥样硬化模型,造模8周;从第3周开始每天灌胃给药1次,连续给药5周。造模结束后应用UPLC-Q-TOF/MS进行血浆样品的分离和数据采集,通过偏最小二乘判别分析法(PLS-DA)分析黄连解毒汤对内源性代谢物的影响;对己鉴定结构的代谢标志物通过Metabo Analyst 4.0进行代谢通路分析。结果筛选和鉴定出35个潜在生物标志物。与模型组比较,黄连解毒汤可显著上调乙内酰脲-5-丙酸、3-羟基-N6,N6,N6-三甲基-L赖氨酸、5-L-谷氨酰基-牛磺酸、3-脱氧肉毒碱、3-羟基-9Z-十八碳烯酰肉毒碱、苏氨酰亮氨酸等20个生物标志物(P <0.05,P <0.01),下调原卟啉IX、17-羟基二十二碳六烯酸、19,20-二羟基-二十二碳五烯酸、肉桂酰甘氨酸、褪黑素等10个生物标志物(P <0.05,P <0.01)。主要影响色氨酸代谢、牛磺酸和亚牛磺酸代谢、组氨酸代谢、赖氨酸降解、卟啉与叶绿素代谢等5条代谢通路;原卟啉IX、乙内酰脲-5-丙酸、3-羟基-N6,N6,N6-三甲基-L赖氨酸、5-L-谷氨酰基-牛磺酸和褪黑素等代谢物是与代谢通路密切相关的关键潜在生物标志物。结论黄连解毒汤对高脂饮食诱导的动脉粥样硬化的治疗作用可能与其改善血浆中内源性代谢物水平,恢复体内正常代谢活动有关。Objective To study the effects of Huanglian Jiedu decoction(HJD)on the potential endogenous markers of experimental atherosclerosis rats by UPLC-Q-TOF/MS,for discover of the potential biomarkers and the molecular mechanism. Methods Male SD rats were randomly divided into five groups,which were the normal control group,model group,and the low-,medium-,high-dose HJD groups(1.5,3,6 g·kg^-1). Except for the normal control group which was fed with normal diet,others groups were fed with high-fat diet,and regularly injected with the vitamin D3. All groups were gavaged once daily from the 3rd week until 8th weeks when they were sacrificed. Then,the blood samples were collected from abdominal aorta,and the plasma were analyzed by UPLC-Q-TOF/MS. Data were processed by PLS-DA. The characterized metabolomics biomarkers were subjected to metaboloic pathway analysis by Metabo Analyst 4.0. Results All groups could be distinguished by metabolomics successfully. Compared with the normal group,35 pathological biomarkers have been changed significantly(P < 0.05,P < 0.01)in the model group. And HJD can significantly increase 20 biomarkers among these 35,including Hydantoin-5-propionic acid,3-Hydroxy-N6,N6,N6-trimethyl-L-lysine,5-L-Glutamyl-taurine,3-Dehydroxycarnitine,3-Hydroxy-9 Zoctadecenoylcarnitine, and so on. It can also significantly decrease 10 biomarkers among the 35, such as Protoporphyrin IX, 17-HDoHE, 19,20-DiHDPA, Cinnamoylglycine, Melatonin, etc(P < 0.05, P < 0.01).According to the metabolic pathways analysis of relevant endogenous markers,it is suggested that HJD may affect the model rats through tryptophan metabolism,taurine and hypotaurine metabolism,histidine metabolism,lysine degradation and porphyrin and chlorophyll metabolism pathways;and five metabolites, including Protoporphyrin IX, Hydantoin-5-propionic acid, 3-Hydroxy-N6,N6,N6-trimethyl-L-lysine, 5-L-Glutamyl-taurine, and Melatonin, were recognized as the key potential biomarkers of the related metabolic pathways. Conclusion This study primarily clarified t

关 键 词：动脉粥样硬化 黄连解毒汤 超高效液相色谱-四极杆-飞行时间质谱 代谢组学 代谢通路 机制 大鼠 

分 类 号：R285.5[医药卫生—中药学]