A Study of 2,3,7,8-Tetrachlorodibenzo-p-dioxin Induced Liver Injury in Jian Carp (Cyprinus carpio var. Jian) Using Precision-Cut Liver Slices

  • Jin-Liang Du
  • Li-Ping Cao
  • Ying-Juan Liu
  • Rui Jia
  • Guo-Jun YinEmail author


The aim of this study was to establish a model for the study of liver injury induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in Jian carp using precision-cut liver slices (PCLS). PCLS were treated with TCDD at concentrations of 0, 0.05, 0.1, 0.3, and 0.6 μg/L for 6 h, followed by collection of the culture supernatant and PCLS for analysis. Several biochemical indices were analyzed, including glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA). Expression of mRNA was also estimated for cytochrome P4501A (CYP1A), aryl hydrocarbon receptor2 (AhR2), and aryl hydrocarbon receptor nuclear translocator2 (ARNT2). Results showed that some significant effects (p < 0.05) in MDA, GSH-Px and PCLS viability were observed at a TCDD concentration as low as 0.05 µg/L, and the observed effects increased with exposure concentration. Following exposure to TCDD for 6 h at a concentration of 0.3 μg/L, significant increases (p < 0.01) in the content of GPT, GOT, MDA, and LDH were observed, while SOD activity, GSH-Px activity, and PCLS viability were decreased (p < 0.01 or p < 0.05). Exposure to 0.3 μg/L TCDD also resulted in increased expression of mRNA for CYP1A, AhR2, and ARNT2. Overall, these results provide evidence of TCDD-induced liver injury and oxidative stress in Jian carp. These results also support the use of PCLS as an in vitro model for the evaluation of hepatotoxicity in Jian carp.


Jian Carp 2,3,7,8-Tetrachlorodibenzo-p-dioxin AhR2 ARNT2 CYP1A 



This study was funded by the National Natural Science Foundation of China (31202002, 31200918), the Central Public-Interest Scientific Institution Basal Research Fund (2013JBFM12), and Jiangsu Science and Technology Department (BK2012535).


  1. Abnet CC, Tanguay RL, Hahn ME, Heideman W, Peterson RE (1999) Two forms of aryl hydrocarbon receptor type 2 in rainbow trout (Oncorhynchus mykiss). Evidence for differential expression and enhancer specificity. J Biol Chem 274:15159–15166CrossRefGoogle Scholar
  2. Bagchi M, Stohs SJ (1993) In vitro induction of reactive oxygen species by 2,3,7,8-tetrachlorodibenzo-p-dioxin, endrin, and lindane in rat peritoneal macrophages, and hepatic mitochondria and microsomes. Free Radic Biol Med 14:11–18CrossRefGoogle Scholar
  3. Baker TR, Peterson RE, Heideman W (2013) Early dioxin exposure causes toxic effects in adult zebrafish. Toxicol Sci 135:241–250CrossRefGoogle Scholar
  4. Bentli R, Ciftci O, Cetin A, Unlu M, Basak N, Cay M (2013) Oral administration of hesperidin, a citrus flavonone, in rats counteracts the oxidative stress, the inflammatory cytokine production, and the hepatotoxicity induced by the ingestion of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Eur Cytokine Netw 24:91–96Google Scholar
  5. Ciftci O, Vardi N, Ozdemir I (2013) Effects of quercetin and chrysin on 2,3,7,8-tetrachlorodibenzo-p-dioxin induced hepatotoxicity in rats. Environ Toxicol 28:146–154CrossRefGoogle Scholar
  6. Collins LL, Williamson MA, Thompson BD, Dever DP, Gasiewicz TA, Opanashuk LA (2008) 2,3,7,8-Tetracholorodibenzo-p-dioxin exposure disrupts granule neuron precursor maturation in the developing mouse cerebellum. Toxicol Sci 103:125–136CrossRefGoogle Scholar
  7. Cravedi JP, Perdu-Durand E, Paris A (1998) Cytochrome P450-dependent metabolic pathways and glucuronidation in trout liver slices. Comp Biochem Physiol C Pharmacol Toxicol Endocrinol 121:267–275CrossRefGoogle Scholar
  8. Dong W, Teraoka H, Yamazaki K, Tsukiyama S, Imani S, Imagawa T, Stegeman JJ, Peterson RE, Hiraga T (2002) 2,3,7,8-Tetrachlorodibenzo-p-dioxin toxicity in the zebrafish embryo: local circulation failure in the dorsal midbrain is associated with increased apoptosis. Toxicol Sci 69:191–201CrossRefGoogle Scholar
  9. Eide M, Karlsen OA, Kryvi H, Olsvik PA, Goksøyr A (2014) Precision-cut liver slices of Atlantic cod (Gadus morhua): an in vitro system for studying the effects of environmental contaminants. Aquat Toxicol 153:110–115CrossRefGoogle Scholar
  10. Elferink MG, Olinga P, Draaisma AL, Merema MT, Bauerschmidt S, Polman J, Schoonen WG, Groothuis GM (2008) Microarray analysis in rat liver slices correctly predicts in vivo hepatotoxicity. Toxicol Appl Pharmacol 229:300–309CrossRefGoogle Scholar
  11. Farmen E, Hultman MT, Anglès D’auriac M, Tollefsen KE (2014) Development of a screening system for the detection of chemically induced DNA methylation alterations in a zebrafish liver cell line. J Toxicol Environ Health A 77:587–599CrossRefGoogle Scholar
  12. Fent K (2001) Fish cell lines as versatile tools in ecotoxicology: assessment of cytotoxicity, cytochrome P4501A induction potential and estrogenic activity of chemicals and environmental samples. Toxicol In Vitro 15:477–488CrossRefGoogle Scholar
  13. Hansson MC, Hahn ME (2008) Functional properties of the four Atlantic salmon (Salmo salar) aryl hydrocarbon receptor type 2 (AHR2) isoforms. Toxicol In Vitro 86:121–130Google Scholar
  14. Hoffman EC, Reyes H, Chu FF, Sander F, Conley LH, Brooks BA, Hankinson O (1991) Cloning of a factor required for activity of the Ah (dioxin) receptor. Science 252:954–958CrossRefGoogle Scholar
  15. Ilavarasi K, Kiruthiga PV, Pandian SK, Devi KP (2011) Hydroxytyrosol, the phenolic compound of olive oil protects human PBMC against oxidative stress and DNA damage mediated by 2,3,7,8-TCDD. Chemosphere 84:888–893CrossRefGoogle Scholar
  16. Jin MH, Hong CH, Lee HY, Kang HJ, Han SW (2010) Toxic effects of lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on development of male reproductive system: involvement of antioxidants, oxidants, and p53 protein. Environ Toxicol 25:1–8CrossRefGoogle Scholar
  17. Kern PA, Fishman RB, Song W, Brown AD, Fonseca V (2002) The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on oxidative enzymes in adipocytes and liver. Toxicology 171:117–125CrossRefGoogle Scholar
  18. Kim HS, Park SY, Yoo KY, Lee SK, Jung WW (2012) Induction of heat shock proteins and antioxidant enzymes in 2,3,7,8-TCDD-induced hepatotoxicity in rats. Korean J Physiol Pharmacol 16:469–476CrossRefGoogle Scholar
  19. Klassen LW, Thiele GM, Duryee MJ, Schaffert CS, Deveney AL, Hunter CD, Olinga P, Tuma DJ (2008) An in vitro method of alcoholic liver injury using precision-cut liver slices from rats. Biochem Pharmacol 76:426–436CrossRefGoogle Scholar
  20. Kleeman JM, Olson JR, Peterson RE (1988) Species differences in 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity and biotransformation in fish. Fundam Appl Toxicol 10:206–213CrossRefGoogle Scholar
  21. Lake BG, Beamand JA, Tredger JM, Barton PT, Renwick AB, Price RJ (1999) Inhibition of xenobiotic-induced genotoxicity in cultured precision-cut human and rat liver slices. Mutat Res 440:91–100CrossRefGoogle Scholar
  22. Lemaire B, Beck M, Jaspart M, Debier C, Calderon PB, Thomé JP, Rees JF (2011) Precision-cut liver slices of Salmo salar as a tool to investigate the oxidative impact of CYP1A-mediated PCB 126 and 3-methylcholanthrene metabolism. Toxicol In Vitro 25:335–342CrossRefGoogle Scholar
  23. Li H, Wang G, Liu S, An Q, Zheng Q, Li B, Li Z (2014) Comparative changes in the antioxidant system in the flag leaf of early and normally senescing near-isogenic lines of wheat (Triticum aestivum L.). Plant Cell Rep 33:1109–1120CrossRefGoogle Scholar
  24. Liu LP, Nie FH, Kong QB, Lin HY, Xie YM, Yang R, Chen JJ (2008) Studies on lipid peroxidation of TCDD in zebrafish. J Guangdong Ocean Univ 28:81–84 (in Chinese) Google Scholar
  25. Liu Q, Rise ML, Spitsbergen JM, Hori TS, Mieritz M, Geis S, Mcgraw JE, Goetz G, Larson J, Hutz RJ, Carvan MJ (2013) Gene expression and pathologic alterations in juvenile rainbow trout due to chronic dietary TCDD exposure. Aquat Toxicol 140:356–368CrossRefGoogle Scholar
  26. Liu YJ, Cao LP, Du JL, Jia R, Wang JH, Xu P, Yin GJ (2015) Protective effects of Lycium barbarum polysaccharides against carbon tetrachloride-induced hepatotoxicity in precision-cut liver slices in vitro and in vivo in common carp (Cyprinus carpio L.). Comp Biochem Physiol C Toxicol Pharmacol 169:65–72CrossRefGoogle Scholar
  27. Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using realtime uantitative PCR and the 2(T)(-Delta Delta C) method. Methods 25:402–408CrossRefGoogle Scholar
  28. Lu M, Chang Z, Bae MJ, Oh SM, Chung KH, Js P (2013) Molecular characterization of the aryl hydrocarbon receptor (AhR) pathway in goldfish (Carassius auratus) exposure to TCDD: the mRNA and protein levels. Fish Shellfish Immunol 35:469–475CrossRefGoogle Scholar
  29. Martnez-Lvarez RM, Morales AE, Sanz A (2005) Antioxidant defenses in fish: biotic and abiotic factors. Rev Fish Biol Fisher 15:75–88CrossRefGoogle Scholar
  30. Miranda CL, Chung WG, Wang-Buhler JL, Musafia-Jeknic T, Baird WM, Buhler DR (2006) Comparative in vitro metabolism of benzo[a]pyrene by recombinant zebrafish CYP1A and liver microsomes from beta-naphthoflavone-treated rainbow trout. Aquat Toxicol 80:101–108CrossRefGoogle Scholar
  31. Ozden S, Catalgol B, Gezginci-Oktayoglu S, Arda-Pirincci P, Bolkent S, Alpertunga B (2009) Methiocarb-induced oxidative damage following subacute exposure and the protective effects of vitamin E and taurine in rats. Food Chem Toxicol 47:1676–1684CrossRefGoogle Scholar
  32. Palaniswamy KS, Vishwanadha VP, Singaravelu SR (2014) Fish oil rich in eicosapentaenoic acid protects against oxidative stress-related renal dysfunction induced by TCDD in Wistar rats. Cell Stress Chaperones 19:409–419CrossRefGoogle Scholar
  33. Park YJ, Lee MJ, Kim HR, Chung KH, Oh SM (2014) Developmental toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in artificially fertilized crucian carp (Carassius auratus) embryo. Sci Total Environ 491–492:271–278CrossRefGoogle Scholar
  34. Peng X, Li F, Li S, Zhu Y (2009) Expression of a mitochondrial gene orfH79 from the CMS-HongLian rice inhibits Saccharomyces cerevisiae growth and causes excessive ROS accumulation and decrease in ATP. Biotechnol Lett 31:409–414CrossRefGoogle Scholar
  35. Schmidt JV, Bradfield CA (1996) Ah receptor signaling pathways. Annu Rev Cell Dev Biol 12:55–89CrossRefGoogle Scholar
  36. Stohs SJ, Al-Bayati ZF, Hassan MQ, Murray WJ, Mohammadpour HA (1986) Glutathione peroxidase andreactive oxygen species in TCDD-induced lipid peroxidation. Adv Exp Med Biol 197:357–365CrossRefGoogle Scholar
  37. Tanguay RL, Abnet CC, Heideman W, Peterson RE (1999) Cloning and characterization of the zebrafish (Danio rerio) aryl hydrocarbon receptor. Biochim Biophys Acta 1444:35–48CrossRefGoogle Scholar
  38. Tohidi F, Cai Z (2015) GC/MS analysis of triclosan and its degradation by-products in wastewater and sludge samples from different treatments. Environ Sci Pollut Res Int 22:11387–11400CrossRefGoogle Scholar
  39. Van De Straat R, De Vries J, Debets AJ, Vermeulen NP (1987) The mechanism of prevention of paracetamol-induced hepatotoxicity by 3,5-dialkyl substitution. The roles of glutathione depletion and oxidative stress. Biochem Pharmacol 36:2065–2070CrossRefGoogle Scholar
  40. Visen PK, Saraswat B, Dhawan BN (1998) Curative effect of picroliv on primary cultured rat hepatocytes against different hepatotoxins: an in vitro study. J Pharmacol Toxicol Methods 40:173–179CrossRefGoogle Scholar
  41. Wang YM, Pu P, Le WD (2007) ATP depletion is the major cause of MPP + induced dopamine neuronaldeath and worm lethality in D-synuclein transgenic C. elegans. Neurosci Bull 23:329–335CrossRefGoogle Scholar
  42. Yu YL, Wei SX, Dong W (2013) Effect of trace dioxin (TCDD) uptake on medaka juvenile spine development. Cereal Feed Ind 12:38–40 (in Chinese) Google Scholar
  43. Zimmermann M, Lampe J, Lange S, Smirnow I, Konigsrainer A, Hann-Von-Weyhern C, Fend F, Gregor M, Bitzer M, Lauer UM (2009) Improved reproducibility in preparing precision-cut liver tissue slices. Cytotechnology 61:145–152CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Jin-Liang Du
    • 1
    • 2
  • Li-Ping Cao
    • 1
    • 2
  • Ying-Juan Liu
    • 3
  • Rui Jia
    • 3
  • Guo-Jun Yin
    • 1
    • 2
    • 3
    Email author
  1. 1.Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research CenterChinese Academy of Fishery SciencesWuxiChina
  2. 2.International Joint Research Laboratory for Fish Immunopharmacology, Freshwater Fisheries Research CenterChinese Academy of Fishery SciencesWuxiChina
  3. 3.Wuxi Fisheries CollegeNanjing Agricultural UniversityWuxiChina

Personalised recommendations