Aquaculture International

, Volume 23, Issue 4, pp 883–893 | Cite as

Hepatoprotective and antioxidant effects of phyllanthin against carbon tetrachloride-induced liver injury in Cyprinus carpio

  • Rui Jia
  • Jin-Liang Du
  • Li-Ping Cao
  • Ying-Juan Liu
  • Pao Xu
  • Guo-Jun Yin


The aim of the present study was to investigate the protective effects and possible mechanism of phyllanthin against carbon tetrachloride (CCl4)-induced hepatic damage in common carp. Fish were fed diet containing four doses of phyllanthin (0, 100, 300, and 600 mg/kg diet) for 60 days, and then given an intraperitoneal injection of CCl4 (30 % in arachis oil) at a dose of 0.5 ml/kg body weight. At 72 h after CCl4 injection, blood and liver samples were collected for the analyses of serum biochemical parameters, peroxidation product, glutathione, and antioxidant enzyme activities. The results showed that administration of phyllanthin at the doses of 300 and 600 mg/kg in the diet prior to CCl4 intoxication considerably inhibited the adverse alterations caused by CCl4 in serum and liver tissue, including glutamate pyruvate transaminase, glutamate oxalate transaminase, lactate dehydrogenase, total protein, albumin, superoxide dismutase, glutathione peroxidase, catalase, glutathione, total antioxidant capacity and malondialdehyde. Furthermore, the positive effects of phyllanthin were confirmed in histopathologic examination. Overall results proved the hepatoprotection of phyllanthin, and it is suggested that phyllanthin may be used as a hepatoprotective agent to prevent liver diseases in fish.


Phyllanthin Carbon tetrachloride Hepatoprotective effect Cyprinus carpio 



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


  1. Ali SS, Kasoju N, Luthra A, Singh A, Sharanabasava H, Sahu A, Bora U (2008) Indian medicinal herbs as sources of antioxidants. Food Res Int 41:1–15CrossRefGoogle Scholar
  2. Banaee M, Sureda A, Mirvaghefi AR, Rafei GR (2011) Effects of long-term silymarin oral supplementation on the blood biochemical profile of rainbow trout (Oncorhynchus mykiss). Fish Physiol Biochem 37:885–896PubMedCrossRefGoogle Scholar
  3. Benzie IF, Strain J (1996) The ferric reducing ability of plasma (FRAP) as a measure of “antioxidant power”: the FRAP assay. Anal Biochem 239:70–76PubMedCrossRefGoogle Scholar
  4. Bhadauria M et al (2009) Emodin reverses CCl4 induced hepatic cytochrome P450 (CYP) enzymatic and ultrastructural changes: the in vivo evidence. Hepatol Res 39:290–300PubMedCrossRefGoogle Scholar
  5. Bhattacharya H, Zhang S, Xiao Q (2008) Comparison of histopathological alterations due to sublethal CCl4 on rosy barb (Puntius conchonius) and amphioxus (Branchiostoma belcheri) with implications of liver ontogeny. Toxicol Mech Methods 18:627–633PubMedCrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  7. Cakmak I, Horst WJ (1991) Effect of aluminium on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycine max). Physiol Plant 83:463–468CrossRefGoogle Scholar
  8. Chandan B et al (2007) Hepatoprotective potential of Aloe barbadensis Mill. against carbon tetrachloride induced hepatotoxicity. J Ethnopharmacol 111:560–566PubMedCrossRefGoogle Scholar
  9. Cheng N, Ren N, Gao H, Lei X, Zheng J, Cao W (2012) Antioxidant and hepatoprotective effects of Schisandra chinensis pollen extract on CCl4-induced acute liver damage in mice. Food Chem Toxicol 23:1235–1241Google Scholar
  10. Cheshchevik V, Lapshina E, Dremza I, Zabrodskaya S, Reiter R, Prokopchik N, Zavodnik I (2012) Rat liver mitochondrial damage under acute or chronic carbon tetrachloride-induced intoxication: protection by melatonin and cranberry flavonoids. Toxicol Appl Pharmacol 261:271–279PubMedCrossRefGoogle Scholar
  11. Ding L, Yongqing M, Fasheng C (1990) Nutritional lipid liver disease of grass carp Ctenopharyngodon idullus (C. et V.). Chin J Oceanol Limnol 8:363–373CrossRefGoogle Scholar
  12. Hernandez-Munoz R et al (1997) Balance between oxidative damage and proliferative potential in an experimental rat model of CCl4–induced cirrhosis: protective role of adenosine administration. Hepatology 26:1100–1110PubMedGoogle Scholar
  13. Huang G-J et al (2013) Hepatoprotective effects of eburicoic acid and dehydroeburicoic acid from Antrodia camphorata in a mouse model of acute hepatic injury. Food Chem 141:3020–3027Google Scholar
  14. Jia R, Cao L, Du J, Xu P, Jeney G, Yin G (2013) The protective effect of silymarin on the carbon tetrachloride (CCl4)-induced liver injury in common carp (Cyprinus carpio). In Vitro Cell Dev Biol Anim 49:155–161Google Scholar
  15. Kandhare AD, Ghosh P, Ghule AE, Zambare GN, Bodhankar SL (2013) Protective effect of Phyllanthus amarus by modulation of endogenous biomarkers and DNA damage in acetic acid induced ulcerative colitis: role of phyllanthin and hypophyllanthin. Apollo Med 10:87–97Google Scholar
  16. Kepekçi RA, Polat S, Çelik A, Bayat N, Saygideger SD (2013) Protective effect of Spirulina platensis enriched in phenolic compounds against hepatotoxicity induced by CCl4. Food Chem 141:1972–1979Google Scholar
  17. Krasnov A, Koskinen H, Rexroad C, Afanasyev S, Mölsä H, Oikari A (2005) Transcriptome responses to carbon tetrachloride and pyrene in the kidney and liver of juvenile rainbow trout (Oncorhynchus mykiss). Aquat Toxicol 74:70–81PubMedCrossRefGoogle Scholar
  18. Krithika R, Mohankumar R, Verma RJ, Shrivastav PS, Mohamad IL, Gunasekaran P, Narasimhan S (2009) Isolation, characterization and antioxidative effect of phyllanthin against CCl4-induced toxicity in HepG2 cell line. Chem Biol Interact 181:351–358PubMedCrossRefGoogle Scholar
  19. Krithika R, Verma RJ, Shrivastav PS, Suguna L (2011) Phyllanthin of standardized phyllanthus amarus extract attenuates liver oxidative stress in mice and exerts cytoprotective activity on human hepatoma cell line. J Clin Exper Hepatol 1:57–67CrossRefGoogle Scholar
  20. Lee C-Y, Peng W-H, Cheng H-Y, Chen F-N, Lai M-T, Chiu T-H (2006) Hepatoprotective effect of Phyllanthus in Taiwan on acute liver damage induced by carbon tetrachloride. Am J Chin Med 34:471–482PubMedCrossRefGoogle Scholar
  21. Lora J, Alonso FJ, Segura JA, Lobo C, Marquez J, Mates JM (2004) Antisense glutaminase inhibition decreases glutathione antioxidant capacity and increases apoptosis in Ehrlich ascitic tumour cells. Eur J Biochem 271:4298–4306PubMedCrossRefGoogle Scholar
  22. Malekinejad H, Alizadeh A, Cheraghi H, Meshkin S, Dardmeh F (2012) The protective effect of liquorice plant extract on CCl4-induced hepatotoxicity in common carp (Cyprinus carpio). Vet res forum 3:158–164Google Scholar
  23. Malhi H, Gores GJ (2008) Cellular and molecular mechanisms of liver injury. Gastroenterology 134:1641–1654PubMedCentralPubMedCrossRefGoogle Scholar
  24. Natsume M et al (1999) Attenuated liver fibrosis and depressed serum albumin levels in carbon tetrachloride-treated IL-6-deficient mice. J Leukoc Biol 66:601–608PubMedGoogle Scholar
  25. Ohkawa H, Ohishi N, Yagi K (1979) Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95:351–358PubMedCrossRefGoogle Scholar
  26. Okafor P, Ikpi M, Uwah I, Ebenso E, Ekpe U, Umoren S (2008) Inhibitory action of Phyllanthus amarus extracts on the corrosion of mild steel in acidic media. Corros Sci 50:2310–2317CrossRefGoogle Scholar
  27. Peskin AV, Winterbourn CC (2000) A microtiter plate assay for superoxide dismutase using a water-soluble tetrazolium salt (WST-1). Clin Chim Acta 293:157–166PubMedCrossRefGoogle Scholar
  28. Pfeifer KF, Weber LJ (1979) The effect of carbon tetrachloride on the total plasma protein concentration of rainbow trout, Salmo gairdneri. Compar Biochem Physiol Part C Compar Pharmacol 64:37–42CrossRefGoogle Scholar
  29. Pikarsky E et al (2004) Pathogenesis of acute viral disease induced in fish by carp interstitial nephritis and gill necrosis virus. J Virol 78:9544–9551PubMedCentralPubMedCrossRefGoogle Scholar
  30. Rajeshkumar N, Joy K, Kuttan G, Ramsewak R, Nair MG, Kuttan R (2002) Antitumour and anticarcinogenic activity of Phyllanthus amarus extract. J Ethnopharmacol 81:17–22PubMedCrossRefGoogle Scholar
  31. Riordan SM et al (2003) Peripheral blood mononuclear cell expression of toll-like receptors and relation to cytokine levels in cirrhosis. Hepatology 37:1154–1164PubMedCrossRefGoogle Scholar
  32. Shaker ME, Zalata KR, Mehal WZ, Shiha GE, Ibrahim TM (2011) Comparison of imatinib, nilotinib and silymarin in the treatment of carbon tetrachloride-induced hepatic oxidative stress, injury and fibrosis. Toxicol Appl Pharmacol 252:165–175PubMedCentralPubMedCrossRefGoogle Scholar
  33. Shivashri C, Rajarajeshwari T, Rajasekar P (2013) Hepatoprotective action of celery (Apium graveolens) leaves in acetaminophen-fed freshwater fish (Pangasius sutchi). Fish Physiol Biochem 39:1057–1069 Google Scholar
  34. Slater TF (1984) Free radical mechanisms in tissue injury. Biochem J 222:1–15Google Scholar
  35. Weber LW, Boll M, Stampfl A (2003) Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. CRC Crit Rev Toxicol 33:105–136CrossRefGoogle Scholar
  36. Wolf JC, Wolfe MJ (2005) A brief overview of nonneoplastic hepatic toxicity in fish. Toxicol Pathol 33:75–85PubMedCrossRefGoogle Scholar
  37. Xia D, Fan Y, Zhang P, Fu Y, Ju M, Zhang X (2013) Protective effects of the flavonoid-rich fraction from rhizomes of Smilax glabra Roxb. on carbon tetrachloride-induced hepatotoxicity in rats. J Membr Biol 246:479–485Google Scholar
  38. Yadav NP, Pal A, Shanker K, Bawankule DU, Gupta AK, Darokar MP, S Khanuja SP (2008) Synergistic effect of silymarin and standardized extract of Phyllanthus amarus against CCl4-induced hepatotoxicity in Rattus norvegicus. Phytomedicine 15:1053–1061PubMedCrossRefGoogle Scholar
  39. Yin GJ, Cao LP, Xu P, Jeney G, Nakao M, Lu CP (2011) Hepatoprotective and antioxidant effects of Glycyrrhiza glabra extract against carbon tetrachloride (CCl4)-induced hepatocyte damage in common carp (Cyprinus carpio). Fish Physiol Biochem 37:209–216. doi: 10.1007/s10695-010-9436-1 PubMedCrossRefGoogle Scholar
  40. Zeng T, Guo F-F, Zhang C-L, Zhao S, Dou D-D, Gao X-C, Xie K-Q (2008) The anti-fatty liver effects of garlic oil on acute ethanol-exposed mice. Chem Biol Interact 176:234–242PubMedCrossRefGoogle Scholar

Copyright information

© Springer International Publishing Switzerland 2014

Authors and Affiliations

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

Personalised recommendations