超高效液相色谱-质谱联用技术在药物肝损伤代谢组学研究中的应用  

Application of ultra high performance liquid chromatographymass spectrometry to metabolomics study of drug-induced hepatotoxicity

作　　者：刘晓燕[1] 刘艳秋[2] 程孟春[1] 肖红斌[1,3] 

机构地区：[1]中国科学院大连化学物理研究所中国科学院分离分析化学重点实验室,辽宁大连116023 [2]大连医科大学中西医结合学院(研究院),辽宁大连116044 [3]北京中医药大学,北京100029

出　　处：《色谱》2015年第7期683-690,共8页Chinese Journal of Chromatography

基　　金：国家科技重大专项(2014ZX09304307-001-006)

摘　　要：药物引起的肝损伤是全世界关注的健康问题,药源性肝损伤的早期诊断仍然是临床治疗肝损伤的一大难题。本研究中,首先利用对乙酰氨基酚、四氯化碳、大黄素、雷公藤甲素和马兜铃酸构建不同类型的肝细胞损伤模型,利用超高效液相色谱-质谱联用技术(UPLC-MS)分别得到正常组和损伤组的细胞代谢轮廓谱。然后利用模式识别构建分类模型,筛选肝损伤相关的差异代谢物。结果显示,不同类型肝损伤在肝细胞代谢谱上可以被区分开,最终鉴定出14种差异代谢物。损伤组肝细胞经保肝阳性药联苯双酯干预后,差异代谢物水平均趋向于正常组,在一定程度上验证了差异标志物与肝损伤的相关性。实验结果表明细胞代谢组学是研究药物肝损伤的有效工具之一。Drug-induced hepatotoxicity is a worldwide health issue. And diagnosing the injury in the early stage is still a challenge in clinic. In this study,pattern recognition analysis of the ultra high performance liquid chromatography-mass spectrometry（ UPLC-MS）of hepatocytes HL7702 was performed to develop differential metabolites related to hepatotoxicity induced by hepatotoxicants,including carbon tetrachloride（CCl4）,acetaminophen（APAP）,emodin, aristolochic acid（ AA）and triptolide. Hepatocytes injuries were induced by 48 h of treatment with CCl4（4 mmol/L）,APAP（6. 5 mmol/L）,emodin（14 μmol/L）,AA（35 μmol/L）and trip-tolide（18 nmol/L）,separately. Global metabolomics profiling,multivariate analysis and data-base searching were performed to discover common differential metabolites for live injury. The positive hepatoprotective drug,bifendate,was used to repair triptolide induced hepatocytes in-jury,and bifendate-induced changes of hepatotoxicity-related metabolites were investigated. In the results,fatty acid oxidation and cellular oxidative stress-related metabolites,including nico-tinamide adenine dinucleotide and glutathione were significantly changed between the control and hepatotoxicant-treated groups,and after treatment with bifendate,those perturbed metab-olites all partly returned to normal level. In conclusion,we discovered potential hepatotoxicity-related metabolites that could be used to evaluate hepatotoxicity induced by chemicals,drugs＆nbsp;and traditional Chinese medicines. This study also proved that metabolomics is one of the effec-tive tools to investigate drug-induced hepatotoxicity.

关 键 词：超高效液相色谱-质谱 代谢组学 肝损伤 

分 类 号：O658[理学—分析化学]