基于1H—NMR代谢组学技术寻找CCl4致大鼠急性肝损伤的代谢标志物

目的采用核磁（1H．NMR）代谢组学技术初步探讨四氯化碳（CCl4）致大鼠急性肝损伤后血清代谢物变化规律。方法SD大鼠随机分为空白、模型两组，模型组ip40％CCl4植物油溶液造成急性肝损伤模型，空白组注射等体积的植物油，造模24h后股动脉取血，常规肝脏病理切片，取血前12h禁食不禁水。比色法测定血清丙氨酸氨基转移酶（ALT）含量，600MHzNMR波谱仪分析血清中小分子代谢物代谢轮廓，并对数据进行多元统计分析。结果模型组中ALT的含量显著升高，肝脏病理切片显示模型组大鼠的肝组织内形成很多空泡，肝细胞变大，坏死严重，有出血和大量中性粒细胞浸润：血清1H-NMR代谢图谱中共鉴定了20个代谢物，并确认了6个与CCl4致肝损伤相关的标志物。结论常规血生化指标和肝病理切片结果与代谢组学的多元统计分析结果相一致，且代谢组学可发现引起肝损伤的代谢标志物。

Searching for biomarkers of CCl4-induced acute liver injury in rats based on 1H-NMR metabonomic approach

Objective The nuclear magnetic resonance （1H-NMR） metabonomic technique is applied to study the change of serum metabolites in Sprague Dawley rats with CCl4-induced acute liver injury. Methods The rats were randomly divided into control and model groups. The rats in the model group were ip injected with 40% CCl4 solution of corn oil, while the rats in the control group were injected with the same amount of corn oil. The blood samples were collected from femoral artery after 24 h of CCl4 treatment. The liver tissues were immediately removed and used for histopathological examination. The alanine aminotransferase （ALT） was determined using commercially available assay kits. The 600 MHz NMR spectrometer was used to analyze the metabolic profile of the small molecule metabolites in serum, and multivariate statistical analysis of the data was also conducted. Results The content of ALT in the model group was significantly increased. Histopathology of liver showed that there were many vacuoles and a large number of neutrophils infiltrating in the liver tissue of model group. The hepatic ceils were necrotic and severely bleeding. Twenty metabolites were confirmed by NMR spectra, and six markers were affirmed to have associations with CCl4-induced liver injury. Conclusion The results concluded by the multivariate statistical analysis of metabonomics are consistent with the conventional biochemical indicators and histopathology of liver. Furthermore, metabonomics could find markers which cause liver injury.