title: 1HNMR-based metabonomic study of sub-chronic hepatotoxicity induced by realgar.
abstract: BACKGROUND
  Realgar has been used as a traditional Chinese medicine (TCM) for thousands of years. Recently, a number of realgar or realgar-containing medicines poisoning cases have been reported. However, the toxicological mechanism of realgar has not been clearly clarified. In present study, the subchronic hepatotoxicity of realgar on mice was investigated using1HNMR-based metabonomic approaches.
  
  
  METHODS
  Twenty-eight male mice were divided into control group and low (0.15g/kg), middle (0.45g/kg), high (1.35g/kg) dosage realgar exposed groups. Their plasma and urine samples were used for NMR spectroscopic metabolic profiling. Principal component analysis (PCA) and pathway analysis were used to detect the hepatotoxic effects of realgar. Liver histopathological examination and plasma clinical chemistry analyses were also performed.
  
  
  RESULTS
  Plasma clinical chemistry analyses showed increased levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein (TP), total cholesterol (TC) and choline esterase (CHE) in realgar-exposed mice indicating liver injury. The PCA score plots showed the metabolic profiles of realgar-exposed mice apparently separated from the controls. Obvious dose-dependent changes of metabolites in urine and plasma of realgar-exposed mice were observed. From the loading plots and boxplots results, the concentrations of VLDL/LDL, 3-HB, lactate, acetate, acetoacetate, creatine, glutamate, methionine, NAc, TMAO, alanine in plasma and pyruvate, succinate, 2-oxoglutarate, DMA, citrate, hippurate, glycine, taurine, phenylalanine, lactate in urine were significantly changed in realgar-exposed mice. The change trends of metabolites in urine and plasma from mice sub-chronic exposed to realgar are similar to those reported in rats acute exposed to realgar, which indicate the acute and sub-chronic toxic mechanism of realgar are same. The disturbed metabolic pathway include energy metabolism, amino acids metabolism and gut bacteria metabolism.
  
  
  CONCLUSIONS
  The present work illustrated the NMR-based metabonomic approach can capture and probe the metabolic alterations induced by traditional Chinese medicine in the toxicological effects.
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Taoguang
authors.initials: T
authors.lastname: Huo
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China; Department of Identification of Traditional Chinese Medicine, School of Pharmacy, Liaoning University of Traditional Chinese Medicine, Shenyang 110032, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Fang
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, PR China.
authors.forename: Longshan
authors.initials: L
authors.lastname: Zhao
authors.affiliation: School of Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, PR China.
authors.forename: Zhili
authors.initials: Z
authors.lastname: Xiong
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Yinghua
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Yanlei
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Cong
authors.initials: C
authors.lastname: Feng
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Mingmei
authors.initials: M
authors.lastname: Yuan
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Shouyun
authors.initials: S
authors.lastname: Wang
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China.
authors.forename: Mo
authors.initials: M
authors.lastname: Chen
authors.affiliation: Department of Health Laboratory Technology, School of Public Health, China Medical University, Shenyang 110122, PR China. Electronic address: jianghong@mail.cmu.edu.cn.
authors.forename: Hong
authors.initials: H
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001152:Arsenicals; D015415:Biomarkers / Q000097:blood / Q000652:urine; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology* / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D004365:Drugs, Chinese Herbal / Q000633:toxicity*; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D055432:Metabolomics / Q000379:methods*; D008813:Mice, Inbred ICR; D015999:Multivariate Analysis; D025341:Principal Component Analysis; D066244:Proton Magnetic Resonance Spectroscopy; D013440:Sulfides / Q000633:toxicity*; D013997:Time Factors; D059550:Toxicity Tests, Subchronic
chemical_list: D001152:Arsenicals; D015415:Biomarkers; D004365:Drugs, Chinese Herbal; D013440:Sulfides; C058317:arsenic disulfide
keywords: Hepatotoxicity; Metabonomics; NMR; Realgar