基于超高效液相色谱-飞行时间质谱技术的肺炎支原体感染小鼠血清代谢组学研究

Serum metabonomics in mice infected with mycoplasma pneumoniae by UPLC-Q-TOF-MS

采用超高效液相色谱与串联四级杆飞行时间质谱(UPLC-Q-TOF-MS)联用技术的代谢组学方法,分析肺炎支原体(MP)感染小鼠血清内源性代谢物的变化,筛选出与肺炎支原体肺炎(MPP)相关的潜在生物标记物,分析代谢通路,探讨MPP的致病机制。采用MP滴鼻感染的方法建立小鼠MPP模型,肺组织病理切片、IgM和支原体核酸的含量测定结果显示造模成功。利用UPLC-Q-TOF-MS方法分析MP感染小鼠的血清代谢轮廓,利用主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)方法进行数据处理。结果显示MP感染模型小鼠的血清代谢谱与正常小鼠有显著差别,经数据库检索、质谱信息匹配,筛选出包括鸟氨酸、皮质醇、维生素A、色氨酸等47种生物标志物,涉及视黄醇代谢、精氨酸与脯氨酸代谢、甾类激素合成等17条代谢通路。该文建立了基于UPLC-Q-TOF-MS技术的MP感染小鼠血清代谢组学研究方法,从整体水平反映了MP感染小鼠内源性小分子的代谢变化,为进一步在分子水平进行药物筛选与评价奠定了基础。

Ultra high performance liquid chromatography coupled with tandem quadrupole time of flight mass spectrometry（UPLC-Q-TOF-MS） was applied to metabonomics study in BALB/c mice infected with mycoplasma pneumoniae（MP） to analyze the changes in serum endogenous metabolites, identify potential biomarkers associated with mycoplasma pneumoniae pneumonia（MPP）, analyze the metabolic pathway and explore the pathogenic mechanism of MPP. The BALB/c mice were inoculated with MP by repeated intranasal infectious routes to establish MPP models, and the results of the lung tissue biopsy, IgM and mycoplasma nucleic acid content determination showed that the models of MP in BALB/c mice were successfully established. UPLC-Q-TOF-MS was used to analyze the serum metabolic profiling of BALB/c mice infected with MP, and then principal component analysis（PCA） was combined with orthogonal partial least squares discriminant analysis（OPLS-DA） for data processing. The results showed that there were significant differences in serum metabolic profile between the MP infected mice and the normal mice. Forty-seven potential biomarkers such as ornithine, cortisol, vitamin A and tryptophan were screened out by database searching and MS information matching. These potential biomarkers related to 17 metabolic pathways including retinol metabolism, arginine and proline metabolism, steroid hormone synthesis and so on. The metabonomic research method for serum of mice infected with mycoplasma pneumoniae based on UPLC-Q-TOF-MS was established in this study. The metabolic changes of endogenous small molecules in mice infected with MP were reflected in the overall level, laying the foundation for the selection and evaluation of MPP drugs.