基于LC-MS的结直肠癌瘀毒内阻证与气血两虚证患者的血清代谢组学研究

Serum metabonomics study of colorectal cancer patients with stasis and poison obstruction syndrome or Qi and blood deficiency syndrome based on LC-MS

目的探讨瘀毒内阻证和气血两虚证结直肠癌患者血清代谢物差异。方法纳入30例结直肠癌患者,分为瘀毒内阻证组(n=17)和气血两虚证组(n=13)。采用液相色谱-质谱联用(liquid chromatography-mass spectrometry,LC-MS)技术检测血清代谢谱,使用主成分分析(principal component analysis,PCA)、偏最小二乘判别分析(partial least squares-discriminant analysis,PLS-DA)和正交-偏最小二乘判别分析(orthogonal partial least squares discriminant analysis,OPLS-DA)进行多元统计分析,并结合受试者操作特征曲线(receiver operating characteristic curve,ROC),计算ROC曲线下面积(area under curve,AUC)值,进一步评估代谢物对证型诊断的敏感度和特异度,对差异代谢物进行层次聚类分析得到热图并通过京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)数据库寻找出共同富集的代谢通路。结果筛选出差异代谢物13种,其中5种显著上调,8种显著下调,其中有2种差异代谢物AUC值大于0.9,除此之外,还有6种差异代谢物AUC值大于0.8。4-O-β-D-吡喃葡萄糖基-D-甘露聚糖、红豆碱、甘露糖、肌苷为气血两虚证敏感代谢物,2-羟基琥珀酸为瘀毒内阻证敏感代谢物。两组的差异代谢物主要富集的信号通路有3个,分别为半乳糖代谢(Galactose metabolism)通路、溶酶体(Lyso⁃some)通路及C型凝集素受体信号通路(C-type lectin receptor signaling pathway)。结论结直肠癌瘀毒内阻证、气血两虚证患者的代谢物存在显著差异,为结直肠癌患者准确、客观的综合征分化和中医治疗提供了可能。

Objectives The purpose of this study was to explore the difference in serum metabolites between patients with colorectal cancer(CRC)with stasis and poison obstruction syndrome or Qi and blood deficiency syndrome.Methods There are thirty patients with CRC were divided into the stasis and poison obstruction syndrome(SPOS group,n=17)and the Qi and blood deficiency syndrome(QBDS group,n=13).Serum metabolic spectrum was detected by liquid chromatography-mass spectrometry,principal component analysis,partial least squares discriminant analysis,and orthogonal partial least squares discriminant analysis,and combined with receiver operating characteristic curve,the area under the curve(AUC)was calculated to further evaluate the sensitivity and specificity of metabolites to syndrome diagnosis.The differential metabolites were analyzed by hierarchical clustering to obtain heat maps,and the co-enriched metabolic pathways were found through the Kyoto Encyclopedia of Genes and Genomes(KEGG)database.Results There are thirteen kinds of differential metabolites were screened,of which five were significantly up-regulated and eight were significantly down-regulated,among which two had AUC values greater than 0.9 and another six had AUC values greater than 0.8.4-O-beta-D-Glucopyranosyl-D-mannose,abrine,D-Mannose,inosine were sensitive metabolites in the QBDS group,and 2-Oxosuccinamate was sensitive metabolite in the SPOS group.There are three signal pathways in which the iconic differential metabolites in these two groups are mainly enriched,namely,Galactose metabolism pathway,Lysosome pathway and C-type lectin receptor signaling pathway.Conclusion There are significant differences in metabolites between the QBDS group and the SPOS group,which provides the possibility for accurate and objective syndrome differentiation and traditional chinese medicine treatment of CRC patients.