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

doi:10.3724/SP.J.1123.2015.04007

Abstract

Abstract:

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 triptolide (18 nmol/L), separately. Global metabolomics profiling, multivariate analysis and database searching were performed to discover common differential metabolites for live injury. The positive hepatoprotective drug, bifendate, was used to repair triptolide induced hepatocytes injury, and bifendate-induced changes of hepatotoxicity-related metabolites were investigated. In the results, fatty acid oxidation and cellular oxidative stress-related metabolites, including nicotinamide adenine dinucleotide and glutathione were significantly changed between the control and hepatotoxicant-treated groups, and after treatment with bifendate, those perturbed metabolites 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 and traditional Chinese medicines. This study also proved that metabolomics is one of the effective tools to investigate drug-induced hepatotoxicity.

Key words: hepatotoxicity, metabolomics, ultra high performance liquid chromatography-mass spectrometry (UPLC-MS)

CLC Number:

O658

LIU Xiaoyan, LIU Yanqiu, CHENG Mengchun, XIAO Hongbin. Application of ultra high performance liquid chromatography- mass spectrometry to metabolomics study of drug-induced hepatotoxicity[J]. Chinese Journal of Chromatography, 2015, 33(7): 683-690.

References

[1] Beger R D, Sun J, Schnackenberg L K. Toxicol Appl Pharm, 2010, 243(2): 154

[2] Fung M. Drug Inf J, 2001, 35(1): 293

[3] Xu X, Qu C Q. Medical Recapitulate (徐鑫, 屈彩芹. 医学综述), 2008, 14(5): 747

[4] Yu X W, Wu Q, Lü W, et al. Chinese Journal of Chromatography (余欣尉, 吴谦, 吕望, 等. 色谱), 2013, 31(7): 691

[5] Li Y J, Zhang C L, Zhang H, et al. Chinese Journal of Chromatography (李艳杰, 张春兰, 张晗, 等. 色谱), 2014, 32(5): 464

[6] Shi D D, Kuang Y Y, Wang G M, et al. Chinese Journal of Chromatography (史栋栋, 况媛媛, 王桂明, 等. 色谱), 2014, 32(3): 278

[7] Peng S Y, Yan L J, Zhang J, et al. Chinese Journal of Chromatography (彭思远, 严丽娟, 张洁, 等. 色谱), 2012, 30(2): 123

[8] Kim K B, Chung M W, Um S Y, et al. Metabolomics, 2008, 4(4): 377

[9] Yang L, Xiong A, He Y, et al. Chem Res Toxicol, 2008, 21(12): 2280

[10] Yamazaki M, Miyake M, Sato H, et al. Toxicol Appl Pharm, 2013, 268(1): 79

[11] Li X, Zhang F, Wang D, et al. J Pharmaceut Biomed Anal, 2014, 89: 42

[12] Kim J W, Ryu S H, Kim S, et al. Anal Chem, 2013, 85(23): 11326

[13] Wang L, Yao D, Chen C. FASEB J, 2013, 27

[14] Xiong Y H, Xu Y, Yang L, et al. J Appl Toxicol, 2014, 34(2): 149

[15] Shaw D. Planta Medica, 2010, 76(17): 2012

[16] Huang T C, Chen S M, Li Y C, et al. Biomed Chromatogr, 2013, 27(9): 1100

[17] Dettmer K, Nuernberger N, Kaspar H, et al. Anal Bioanal Chem, 2011, 399(3): 1127

[18] Tautenhahn R, Patti G J, Rinehart D, et al. Anal Chem, 2012, 84(11): 5035

[19] Chen J, Zhao X, Fritsche J, et al. Anal Chem, 2008, 80(4): 1280

[20] Shen G, Chen Y, Sun J, Zhang R, et al. J Proteome Res, 2011, 10(4): 1953

[21] Chan E C Y, Pasikanti K K, Nicholson J K. Nat Protoc, 2011, 6(10): 1483

[22] Peng S, Yan L, Zhang J, et al. J Pharmaceut Biomed Anal, 2013, 86: 56

[23] Sakagami H, Sugimoto M, Tanaka S, et al. Metabolomics, 2014, 10(2): 270

[24] Amet Y, Zerilli A, Goasduff T, et al. Biochem Pharmacol, 1997, 54(8): 947

[25] Matsunaga I, Sumimoto T, Ueda A, et al. Lipids, 2000, 35(4): 365

[26] Amet Y F, Adas A A. Alcohol Clin Exp Res, 1998, 22(7): 1493

[27] Ana Carla B B, Fabricio B, Livia B, et al. Acta Pharmacol Sin, 2009, 30(1): 90

[28] Liu T F, Vachharajani V T, Yoza B K, et al. J Biol Chem, 2012, 287(31): 25758

[29] Diaz Ochoa J G, Bucher J, Pery A R R, et al. Front Pharmacol, 2012, 3: 204

[30] Pan S Y, Yang R, Dong H, et al. Eur J Pharmacol, 2006, 552(1/3): 170