摘要
深静脉血栓(DVT)是一种血栓栓塞性疾病,具有高发病率、高死亡率和高后遗症3大特点。采用左股静脉不完全结扎加高渗盐水刺激建立DVT大鼠模型,使用超高效液相色谱-静电场轨道阱高分辨质谱(UHPLC-Orbitrap HRMS)检测假手术组与DVT模型组的血浆代谢谱,用主成分分析(PCA)及正交偏最小二乘-判别分析(OPLS-DA)对代谢组数据进行多元统计分析,观察两组间的代谢表型差异,将多变量模型分析中的变量重要性值(VIP>1)以及代谢物在模型组中的变化倍数(FC≤0.5或FC≥2,且P<0.05)作为差异代谢物筛选条件。最终在DVT模型组与假手术组间筛选得到27种差异代谢物,这些代谢物反映了DVT大鼠的代谢紊乱情况。具体表现为与假手术组相比,DVT模型组中三甲基胺氮氧化物(TMAO)、维生素K、鹅去氧胆酸、牛磺酸、1-甲基烟酰胺、7-酮胆固醇、反式十六烷基-2-烯醇肉碱、乙烯基乙酰甘氨酸、丙酰脯氨酸、咪唑乙酸、咪唑乙酸核糖苷、1,3,7-三甲基尿酸、1-丁胺、2-羟基异丙酸、吡哆醛、5α-四氢皮质酮、苯乳酸的水平显著升高;而3-脱氢肉碱、磷脂酰胆碱22∶6/20∶2(PC 22∶6/20∶2)、甘油二酯18∶3/20∶4(DG 18∶3/20∶4)、溶血磷脂酰胆碱20∶2(LysoPC 20∶2)、波维酸、鹅肌肽、L-肌肽、辛酸、羟基丙酮酸、3-羟基癸酸的水平显著降低。基于京都基因与基因组百科全书数据库(KEGG)代谢通路的差异丰度(DA)分析显示DVT模型大鼠与假手术组的代谢通路差异主要集中在初级胆汁酸生物合成、胆汁分泌、组氨酸代谢、亚油酸代谢、甘油磷脂代谢和β-丙氨酸代谢。紊乱的代谢物和代谢途径可为进一步深入理解DVT的病理机制、寻找诊断标志物及药物作用靶点提供参考。
Deep vein thrombosis(DVT)is a venous thromboembolic disease characterized by high incidence,mortality,and sequelae.Therefore,the effective prevention of DVT has become a critical public health concern.However,due to its complexity,the pathophysiological mechanism of DVT remains unclear.Metabolomics can be employed to analyze disease characteristics and provide scientific evidence on the underlying mechanisms.In this study,an established left femoral vein ligation rat model of DVT(n=10)was used and compared with sham surgery controls(n=10).In the DVT group,rats were anesthetized using an intraperitoneal injection of 10%chloral hydrate(300 mg/kg),after which the hair was shaved and the groin disinfected.A 2-cm longitudinal incision was made along the midpoint of the left groin area,and then the left femoral vein was separated.The vein was partially ligated at its proximal end to shrink the blood vessel lumen to approximately half.Then,0.4 mL of 10%hypertonic saline was slowly injected from the distal end of the left femoral vein.At the same time,the femoral vein turned dark red,which indicated the formation of thrombosis.Finally,the incision was sutured after verifying bleeding in the surrounding tissue.Keeping all other procedures the same as the DVT group,the vein in the control group was not ligated or stimulated using hyper-tonic saline.The abdominal aorta plasma from rats in each group was collected seven days later.Untargeted metabolomics analysis based on ultra-high performance liquid chromatography-electrostatic field orbitrap high resolution mass spectrometry(UHPLC-Orbitrap HRMS)was conducted to investigate the plasma metabolic profiles of the sham surgery control and DVT groups.Principal component analysis(PCA)and orthogonal to partial least squares discrimi-nant analysis(OPLS-DA)on metabolome data for multivariate statistical analysis were employed to assess differences in the metabolic profile between the two groups.The results revealed distinct profiles for the DVT and control groups.The selection crit
作者
谷艳
臧鹏
李进霞
闫燕艳
王佳
GU Yan;ZANG Peng;LI Jinxia;YAN Yanyan;WANG Jia(School of Medicine, Shanxi Datong University, Datong 037009, China;Datong No. 3 People’s Hospital, Datong 037000, China)
出处
《色谱》
CAS
CSCD
北大核心
2022年第8期736-745,共10页
Chinese Journal of Chromatography
基金
山西省科技厅应用基础研究项目(201901D211429).
关键词
超高效液相色谱-静电场轨道阱高分辨质谱
代谢组学
深静脉血栓
ultra-high performance liquid chromatography-electrostatic field orbitrap high resolution mass spectrometry(UHPLC-Orbitrap HRMS)
metabolomics
deep vein thrombosis(DVT)
