基于GC-TOFMS的虚寒证原发性痛经患者尿液代谢组学

目的: 比较虚寒证原发性痛经患者与正常组尿液内源性代谢物的差异,从整体角度阐释其病理生理学机制,深化中医整体观在现代实验研究中的应用。 方法: 采用病证结合模式,基于气相色谱-飞行时间质谱(GC-TOFMS)技术,以N-甲基-N-三甲基-硅基三氟乙酰胺(MSTFA)作为衍生化试剂,检测虚寒证原发性痛经患者及正常组月经周期第2天(MC₂)尿液代谢产物,得出两组总离子流色谱图(TIC),结合非监督的主成分分析(PCA)进行多维统计分析,寻找两组的差异性代谢产物。 结果: 虚寒组与正常组比较,差异代谢产物有果糖、麦芽糖、阿拉伯呋喃糖、异柠檬酸、乳酸、马尿酸、甘氨酸、丝氨酸、苏氨酸、软脂酸、硬脂酸、原藻醇12种。与正常组比较,虚寒组升高的有乳糖,降低的有果糖、麦芽糖、阿拉伯呋喃糖、异柠檬酸、马尿酸、甘氨酸、丝氨酸、苏氨酸,软脂酸、硬脂酸、原藻醇。 结论: 虚寒证原发性痛经组与正常组尿液的差异代谢物质涉及到糖、氨基酸、脂酸代谢及肠道微生物菌群。

Asthenia-cold Syndrome in Patients with Primary Dysmenorrhea Based on Urine Metabolomics

Objective:To compare the differences between the asthenia-cold syndrome in patients with primary dysmenorrhea and normal group in urinary endogenous metabolites, and elaborate the path physiological mechanisms of the asthenia-cold syndrome in patients with primary dysmenorrheal. Method: N-methyl-N-(trimethylsilyl) tri-fluoroacetamide (MSTFA) was selected as derivatization reagent to detect urine metabolites at the second day in menstrual cycle (MC₂) of the two groups using gas chromatography-time of flight mass spectrometer (GC-TOFMS), drawn the total ion chromatogram(TIC) of each group, combination with unsupervised principal component analysis (PCA) for the multi-dimensional statistical analysis to look for variability material between the two groups. Result: Compared with the normal group, twelve variability materials in asthenia-cold group were detected. They were fructose, maltose, arbinofuranose, isocitric acid, lactic acid, hippuric acid, glycine, serine, threonine, palmitic acid, octadecanoic acid, and erythritol. Among the total materials only lactic acid in asthenia-cold group was higher than in the normal group;the rest materials in asthenia-cold group were all lower than in the normal group. Conclusion: Asthenia-cold syndrome of the primary dysmenorrheal patients and the normal group suggested significant difference in urine metabolites. These variability materials are related to several metabolic pathways such as glycolmetabolism, amino acid metabolism, fatty acid metabolism and disturbance of enterobacteria.