儿童哮喘患者中医证型的代谢组学研究

Metabolomics Research on TCM Syndrome of Childhood Asthma

目的:基于气相色谱/质谱(GC/MS)技术的代谢组学方法,分析儿童哮喘两种中医证型患者尿液中的内源性小分子代谢物,探索两种中医证型分类的物质基础。方法:采用GC/MS技术检测儿童哮喘肺脾气虚型、气阴两虚型患者以及正常儿童的尿液中小分子代谢物,利用SIMCA-P软件进行主成分分析(principal component analysis,PCA)及正交偏最小二乘法(OPLSA)分析,研究尿液中小分子代谢物在各组间差异。结果:儿童哮喘肺脾气虚型、气阴两虚型患者代谢谱具有良好的分离趋势,并与正常儿童对照组有明显区分。利用代谢物谱库(如Wiley和NIST质谱库)及本实验室建立的标准品代谢物谱库鉴定了17个差异代谢物。儿童哮喘两种"证型"患者间尿液中丁酸、磷酸盐、L-苏氨酸、肌酐、甘氨酸、D-葡萄糖、马尿酸、假尿苷的相对含量存在显著差异,这些代谢物与能量代谢、肠道菌群代谢、氨基酸代谢和核苷酸代谢异常相关,它们是区分这两种中医证型的潜在生物标志物。结论:代谢组学技术能够全面地反映肺脾气虚型、气阴两虚型儿童哮喘的代谢共性和各自特征,代谢组学所研究的体内小分子代谢物可能是中医分型的物质基础。

Objective ： Based on gas chromatography/mass spectrometry （GC/MS） metabolomics techniques, endoge- nous small molecule metabolites of urine in asthma children with two TCM syndrome was analyzed and the material basis of the two syndromes type classification was explored. Methods ： Small molecule metabolites in the urine of childhood asth- ma with lung and spleen deficiency syndrome and Qi - Yin deficiency syndromes as well as normal children were detected with GC/MS. The principal component analysis （PCA） and Orthogonal -Projection to Latent Structures （OPLS） were analyzed using SIMCA - P software. The differences of small molecule metabolites in urine among the groups were studied Results Metabolic profile appeared good separation trend between childhood asthma with lung and spleen deficiency syn- drome and Qi -Yin deficiency syndromes. Three groups were distinguished clearly. Seventeen differences in metabolites were identified using metabolite library （ such as Wiley and NIST mass spectral library） and the establishment of the labo- ratory~ standard library metabolites. There were significant differences about urine relative content of butyric acid,phos- phate, L - threonine, creatinine, glycine, D - glucose, hippuric acid, the pseudouridine between children asthma with two kinds of syndromes. The dysfunction of energy metabolism,intestinal flora metabolism, amino acid metabolism and nucleo- tide metabolism were related to these metabolites. They were the potential biomarkers for distinguishing two kinds of TCM syndromes. Conclusion： Metabolomic technology can fully reflect their common type and metabolic characteristics between childhood asthma with lung and spleen deficiency syndrome and Qi - Yin deficiency syndrome. The small molecule me- tabolites in vivo based on metabolomic study may be the material basis of TCM syndrome.