二乙基亚硝胺诱导的肝癌模型大鼠尿液代谢组学研究

Metabonomics study of urine in rat model with hepatocellular carcinoma induced by diethylnitrosamine

目的应用核磁共振检测法(~1H-NMR)对大鼠尿液小分子代谢标志物及其参与的代谢通路进行分析。方法通过饮用质量浓度为0.1 mg·mL^(-1)的二乙基亚硝胺溶液(DEN)造肝癌大鼠模型,同时用饮用灭菌水的大鼠作为正常对照组,18周后采集尿样并进行核磁共振检测,在显微镜下对肝组织进行观察对比。结果模型组大鼠尿液中异亮氨酸、乳酸、丙氨酸、醋酸盐、糖蛋白、丙酮酸盐、琥珀酸盐、α-酮戊二酸、苯丙氨酸和甲酸盐10种代谢物的含量显著降低;而柠檬酸、二甲基亚硝酸、肌酸、牛磺酸和肌酸酐5种代谢产物的含量显著增加。采用MetaboAnalyst 3.0对差异性代谢物进行分析,发现共有4种代谢通路发生了明显变化。结论通过对大鼠尿液代谢物进行检测分析,为肝癌提供早期诊断依据。

Objective To analyze the metabolic markers of small molecules in urine and the metabolic pathways involved in the rat by hydrogen nuclear magnetic resonance (1 H-NMR) detection. Methods Rat models of hepatocellular carcinoma were established by drinking diethylnitrosamine (DEN) solution (0.1 mg·mL -1 ),and rats were treated with sterilized water as normal control group.After18 weeks, the urine samples were collected and analyzed by 1 H-NMR.The liver tissue was observed under microscope. Results In model group,10 metabolites,including isoleucine,lactic acid,alanine,acetate, glycoprotein,pyruvate,succinate,α-ketoglutaric acid, phenylalanine and formate were significantly decreased;the contents of citric acid,dimethylnitrosamine,creatine,taurine and creatinine were increased significantly.MetaboAnalyst 3.0 was used to analyze the different metabolites and 4 metabolic pathways were found changed significantly. Conclusion The detection and analysis of urinary metabolites in rats provide an early diagnostic basis for hepatocellular carcinoma.