冠心病痰瘀互结证小型猪血浆代谢组学研究  

Metabolomics Study on Plasma of Miniature Pigs with Phlegm-stasis Syndrome of Coronary Heart Disease

作　　者：张国瑗 李磊[1] 孟红旭[1] 王奥奥 王紫艳 胡广 李瑛 马彦雷[1] 史跃[1] 孙明谦[1] 刘建勋[1] ZHANG Guoyuan;LI Lei;MENG Hongxu;WANG Aoao;WANG Ziyan;HU Guang;LI Ying;MA Yanlei;SHI Yue;SUN Mingqian;LIU Jianxun(Institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,中药药理北京市重点实验室,北京100091

出　　处：《中国中医药信息杂志》2023年第6期117-122,共6页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家自然科学基金(82030124、81873041);中国中医科学院科技创新工程项目(CI2021A01707);国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202007)。

摘　　要：目的基于液相色谱-质谱联用技术对冠心病痰瘀互结证小型猪血浆除磷脂外的代谢物进行代谢组学分析。方法将小型猪分为对照组和模型组,采用高脂饲料喂养结合冠状动脉内皮损伤法建立冠心病痰瘀互结证小型猪模型。HE和Masson染色观察心肌组织病理变化;血浆样本用96孔SPE板进行除蛋白和去磷脂处理,采用超高效液相色谱-四级杆飞行时间质谱(UPLC-QTOF-MS)对样本进行检测,偏最小二乘判别分析(PLS-DA)进行数据分析,通过HMDB数据库和二级质谱裂解规律比对,对内源性成分进行定性分析,筛选差异代谢物,通过MetaboAnalyst 5.0结合HMDB、KEGG数据库对差异代谢物进行代谢通路分析。结果HE和Masson染色显示,模型组心肌细胞肥大,部分心肌细胞融合,心肌间质水肿,大量胶原纤维增生;PLS-DA结果显示,模型组和对照组血浆代谢物差异明显,筛选出7a,12a-二羟基-3-氧代-4-胆氨酸、3b,12a-二羟基-5a-胆酸、脱氧胆酸甘氨酸结合物、别石胆酸、十六烷二酸、3-氧代十八烷酸、二十二碳六烯酸、丁烯肉碱、3-羟基-9-十六碳烯酰肉碱、3-羟基十六烷基肉碱、(9E)-10-硝基十八烷-9-烯酰肉碱等26个差异代谢物,涉及肉碱穿梭代谢、花生四烯酸代谢、胆汁酸生物合成、胆碱代谢、丙酮酸代谢等途径。结论内源性非磷脂类成分如胆酸类、脂肪酸类、氨基酸类、酰基肉碱类及其生物代谢途径参与冠心病痰瘀互结证发病过程。本研究可为了解冠心病痰瘀互结证病程进展提供实验依据。Objective To analyze the metabolomic characteristics of plasma of miniature pigs with phlegm-stasis syndrome of coronary heart disease based on liquid chromatography-mass spectrometry.Methods The miniature pigs were divided into control group and model group.The model of phlegm-stasis syndrome of coronary heart disease in miniature pigs was established by high-fat feed combined with coronary artery balloon strain endothelium.Myocardial pathological changes were observed with HE and Masson staining;plasma samples were treated with SPE 96-well plates to remove proteins and phospholipids,and ultra-high performance liquid chromatography-quadrupole time-offlight mass spectrometry(UPLC-QTOF-MS)was used for sample detection,partial least squares discriminant analysis was used for data analysis,qualitative analysis of endogenous components and screening of differential metabolites were carried out through comparison of HMDB database and secondary mass spectrometry,and MetaboAnalyst 5.0 combined with KEGG and HMDB databases was used to analyze metabolic pathways of differential metabolites.Results HE and Masson staining showed that myocardial cells in model group were hypertrophy,some myocardial cells were fused,myocardial interstitial edema,and a large number of collagen fibers were proliferated;PLS-DA results showed that there were significant differences in plasma metabolites between model group and control group,and 26 different metabolites were screened,including 7a,12a-dihydroxy-3-oxo-4-choline,3b,12a-dihydroxy-5a-cholic acid,deoxycholic acid glycine conjugate,allocholic acid,hexadecane diacid,3-oxodectanoic acid,docosahexaenoic acid,butene carnitine,3-hydroxy-9-hexadecenyl carnitine,3-hydroxyhexadecyl carnitine,(9E)-10-nitrooctadec-9-enoylcarnitine,etc.,involving carnitine shuttle metabolism,arachidonic acid metabolism,bile acid biosynthesis,choline metabolism,pyruvate metabolism and other pathways.Conclusion Endogenous non phospholipid components,such as bile acids,fatty acids,amino acids,acyl carnitines and their

关 键 词：冠心病 痰瘀互结证 代谢组学 超高效液相色谱-四级杆飞行时间质谱 小型猪 

分 类 号：R285.5[医药卫生—中药学]