非酒精性脂肪性肝病肝胆湿热证患者尿液代谢组学研究

Study on urine metabolomics of liver and gallbladder damp-heat syndrome patients with nonalcoholic fatty liver

目的:研究非酒精性脂肪性肝病(NAFLD)肝胆湿热患者尿液中代谢物的变化,寻找其特征代谢物,探索NAFLD肝胆湿热证的证候本质。方法:采用气相色谱质谱联用(GC-MS)技术检测NAFLD肝胆湿热证(142例)、无证可辨(61例)以及健康对照组(98名)的尿液中小分子代谢产物。采用主正交偏最小二乘法(OPLS)的方法,寻找并鉴定肝胆湿热证患者尿液特征代谢物。结果:无证可辨组与健康对照组、肝胆湿热组与无证可辨组的代谢谱有很好的区分,获得了12个有显著性差异的代谢物,包括4-硝基苯甲酸酯、异龙脑、1H-吲哚、醋酸、色氨酸、古龙酸、岩藻糖、尿嘧啶、假尿苷、2,4-二氨基丁酸、葡萄糖、油酸。这12个代谢物可以作为NAFLD肝胆湿热证诊断的尿液中潜在的生物标志物;色氨酸代谢、赖氨酸生物合成、糖酵解或糖异生3条代谢通路与NAFLD肝胆湿热证密切相关。结论:NAFLD肝胆湿热证的生物学本质体现在色氨酸代谢、赖氨酸生物合成、糖酵解或糖异生等方面,基于GC-MS结合模式识别的代谢组学方法有望揭示中医证候本质的科学内涵。

Objective:To study the changes of metabolites in urine of liver and gallbladder damp-heat syndrome patients with non-alcoholic fatty liver,and search for characteristic metabolites and explore the syndrome nature of liver and gallbladder damp-heat syndrome patients with non-alcoholic fatty liver.Methods:Gas chromatography-mass spectrometry(GC/MS)was used to detect small molecule metabolites in the urine of non-alcoholic fatty liver with liver and gallbladder damp-heat syndrome patients(142 cases),unidentifiable syndrome patients(61 cases)and healthy controls(98 cases).The main orthogonal partial least squares(OPLS)method was used to find and identify characteristic metabolites in urine of liver and gallbladder dampheat syndrome patients with non-alcoholic fatty liver.Results:The metabolic profiles of the unidentifiable group and the healthy group were well distinguished;the metabolic profiles of the liver and gallbladder damp-heat group were well distinguished from the unidentifiable group.Twelve significantly different metabolites were obtained including 4-nitrobenzoate,isoborneol,1 H-indole,acetic acid,tryptophan,colic acid,fucose,uracil,pseudouridine,2,4-diaminobutyric acid,glucose,oleic acid,etc.These 12 metabolites can be used as potential biomarkers in the diagnosis of liver and gallbladder damp-heat syndrome patients with non-alcoholic fatty liver.Tryptophan metabolism,lysine biosynthesis,glycolysis,or gluconeogenesis are closely related to liver and gallbladder damp-heat syndrome patients with non-alcoholic fatty liver.Conclusion:The biological essence of liver and gallbladder damp-heat syndrome patients with non-alcoholic fatty liver is embodied in the aspects of tryptophan metabolism,lysine biosynthesis,glycolysis or gluconeogenesis.A metabolomic method based on GC-MS combined with pattern recognition is expected to reveal the scientific connotation of the essence of TCM syndromes.