利用液相色谱质谱联用技术开展维医异常黑胆质证哮喘的血清代谢组学研究,血清样品经4倍体积乙腈沉淀蛋白处理,对采集的谱图进行峰提取、峰匹配和缺失值过滤后,采用主成分分析和偏最小二乘-判别分析进行数据解读。结果发现,异常黑胆质证哮喘病人的代谢表型发生了显著变异,主成分分析得分散点图中健康对照组与患病组分类良好;偏最小二乘-判别分析发现了15个表征代谢表型变异的生物标记物,对其中9个进行了结构鉴定,预测了其余6个的分子式。与健康对照组比较,肉豆蔻酰溶血磷脂酰胆碱、十五酰溶血磷脂酰胆碱、棕榈酰溶血磷脂酰胆碱、二十碳三烯酰溶血磷脂酰胆碱、油酰溶血磷脂酰胆碱、十七酰溶血磷脂酰胆碱、单棕榈酸甘油酯、N-十四酰二氢鞘氨醇和未知代谢物C20H43NO2、C22H47NO3在患病组的含量显著下降,而硬脂酰胺和未知代谢物C16H35NO、C16H33NO、C27H3O14、C20H39NO的含量显著升高。异常黑胆质证哮喘中溶血磷脂酰胆碱的降低可能与炎症介导的肺表面活性物质的功能异常有关,这为进一步认识维医异常黑胆质的发病机制与维医独特的治疗原则提供了初步基础。
Serum metabonomics investigation into asthma diagnosed as abnormal savda syndrome in Uighur Traditional Medicine was carried out with HPLC/Q-TOF MS. Prior to LC-MS analysis, a deproteination method with four timesvolume acetonitrile was employed to prepare serum samples. A multi-steps data preprocessing procedure including peak extraction, chromatographic alignment and missing value filtering was completed, and then principal component analysis (PCA) and partial least square-discriminant analysis (PLS-DA) were fulfilled to distinguish the metabolic phenotypes and pinpoint potential biomarkers. It was found that the metabolic phenotype in patient group underwent significant variation, which resulted in the obvious classification between the health and patient groups in the score plot of PCA model. The influential biomarkers accounting for the metabolic signature perturbation were discovered in the PLS-DA model, of which nine biomarkers were tentatively structure identified and the formula of the other six ones were also postulated in accordance with exact mass. The findings demonstrated that the expression of myristoyl- lysophosphatidylcholine (lysoPC), pentadecanoyl-lysoPC, palmitoyl-lysoPC, eicosatrienoyl-lysoPC, oleoyl-lysoPC, heptadecanoyl-lysoPC, monopalmitin, N-tetradecanoyl-sphinganine and unknown metabolites C20H43NO2, C22H47NO3 significantly decreased in patient group comparing with health group, whereas, the expression of stearamide and other four unknown metabolites C16H35NO, C16H33NO, C27H45N3014, C20H39NO significantly elevated. It was inferred that the decreased expression of lysoPCs in patient group might in part correlate with pulmonary surfactant dysfunction mediated by inflammation reaction. These findings would facilitate further understanding of pathogenesis of abnormal savda syndrome, and serve as primary groundwork for elucidating the unique therapeutic principles in Uighur Traditional Medicine.