色谱 ›› 2019, Vol. 37 ›› Issue (1): 71-79.DOI: 10.3724/SP.J.1123.2018.09008

• 研究论文 • 上一篇    下一篇

基于超高效液相色谱-四极杆飞行时间质谱联用技术的血瘀模型大鼠血浆代谢组学分析

杨秀娟1,2, 杨志军1,2, 李硕1,2, 邓毅1,2, 杨延泽1, 曼琼1, 李鹏杰1   

  1. 1. 甘肃中医药大学药学院, 甘肃 兰州 730000;
    2. 甘肃省高校中(藏)药化学与质量研究省级重点实验室, 甘肃 兰州 730000
  • 收稿日期:2018-09-05 出版日期:2019-01-08 发布日期:2014-11-29
  • 通讯作者: 邓毅,Tel:(0931)5109886,E-mail:dengyi@gszy.edu.cn.
  • 基金资助:

    国家自然科学基金地区科学基金项目(81360633);甘肃省自然科学基金项目(1506RJZA044);甘肃省中管局项目(GZK-2016-25);甘肃省教育厅高等学校科研项目(2017A-056,2016B-055);2016-度甘肃省高校中(藏)药化学与质量研究省级重点实验室开放基金项目(zzy-2016-06);敦煌医学与转化教育部重点实验室开放基金项目(DHYX18-11).

Rat plasma metabolomics in blood stasis model based on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry

YANG Xiujuan1,2, YANG Zhijun1,2, LI Shuo1,2, DENG Yi1,2, YANG Yanze1, MAN Qiong1, LI Pengjie1   

  1. 1. Gansu College of Traditional Chinese Medicine, Lanzhou 730000, China;
    2. Key Laboratory of Chemistry and Quality for Traditional Chinese Medicine of Gansu Province, Lanzhou 730000, China
  • Received:2018-09-05 Online:2019-01-08 Published:2014-11-29
  • Supported by:

    National Natural Science Foundation of China, Regional Science Foundation Project (No. 81360633); Gansu Provincial Natural Science Foundation of China (No. 1506RJZA044); Research Project Fund Project of Gansu Provincial Administration of Traditional Chinese Medicine (No. GZK-2016-25); Scientific Research Project of the Higher Education Institutions of Gansu Province (Nos. 2017A-056, 2016B-055); Research Fund Project of Key Laboratory of Chemistry and Quality for Traditional Chinese Medicine of Gansu Province (No. zzy-2016-06); Research Fund Project of Key Laboratory for Transfer of Dunhuang Medicine at the Provincial and Ministerial Level (No. DHYX18-11).

摘要:

采用盐酸肾上腺素加冰水浴建立急性血瘀大鼠模型,使用超高效液相色谱-四极杆飞行时间质谱(UPLC-Q-TOF/MS)检测空白对照组与血瘀模型组中血浆代谢物,用主成分分析(PCA)、有监督偏最小二乘法判别分析(PLS-DA)及正交偏最小二乘法判别分析(OPLS-DA)对代谢组学数据进行多维统计分析,筛选潜在生物标志物。与对照组相比,在血瘀模型组大鼠血浆中检测出46个差异代谢物,血瘀模型组中乙酰胆碱、N6,N6,N6-三甲基-L-赖氨酸、胞嘧啶、乙酰肉碱等21个代谢物显著上调,吲哚丙酸、LysoPC(14:0)等25个代谢物显著下调,可能与脂质代谢、半乳糖代谢、亚油酸代谢、不饱和脂肪酸生物合成、糖酵解、花生四烯酸代谢等通路有关。代谢产物可作为血瘀证研究中的重要标记物,该研究结果有助于揭示血瘀证的发病机制,可为临床血瘀疾病的诊断及选用药物治疗提供思路,为后续治疗手段提供参考依据。

关键词: 超高效液相色谱-四极杆飞行时间质谱, 代谢物, 代谢组学, 机制, 血瘀证

Abstract:

Acute blood stasis syndrome was induced in rats by adrenaline hydrochloride and ice water. Ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was conducted on plasma metabolites of normal and model rats. Principal component analysis (PCA), differentiation analysis of supervised partial least squares method (PLS-DA), and orthogonal to partial least squares discriminant analysis (OPLS-DA) on metabolomics data for multidimensional statistical analysis were employed, and the resulting biomarkers were screened. Compared to the normal group, there were 46 endogenous metabolites in blood stasis-rat plasma. Of these, 21 metabolites were significantly upregulated, such as acetylcholine, N6,N6,N6-trimethyl-L-lysine, cytosine, and acetylcarnitine, while 25 metabolites were reduced, including indoleacrylic acid, and lysoPC(14:0). These metabolites were mainly related to metabolic pathways, including lipid metabolism, galactose metabolism, linoleic acid metabolism, biosynthesis of unsaturated fatty acids, glycolysis, and arachidonic acid metabolism. In conclusion, these results indicated that metabolites could be used as important biomarkers for blood stasis syndrome, and could help in revealing the mechanism of blood stasis disease and provide a reference network to determine the disease development stage and appropriate follow-up treatment. Studying altered metabolites in blood stasis model rats can provide insights useful for the diagnosis of blood stasis in the clinic and for the development of drug therapies.

Key words: blood stasis syndrome, mechanism, metabolites, metabolomics, ultra performance liquid chromatography-quadrupole-time-of-flight mass spectro-metry (UPLC-Q-TOF/MS)

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