Skip to main content
SpringerLink
Log in

Protective action of the phyllanthin against carbon tetrachloride-induced hepatocyte damage in Cyprinus carpio

  • Report
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

The present study was to investigate the protective effect and possible mechanism of phyllanthin against carbon tetrachloride (CCl4)-induced hepatocyte damage in carp. Phyllanthin (5, 10, and 15 μg/ml) was added to carp primary hepatocytes before (pre-treatment) and after (post-treatment) incubation of the hepatocytes in medium containing CCl4 at 8 mM; supernatant and cell were collected for the analyses of cell viability, biochemical parameters, and gene expression. The results showed that phyllanthin at the concentration of 15 μg/ml significantly suppressed the elevation of glutamate pyruvate transaminase (GPT), glutamate oxalate transaminase (GOT), lactate dehydrogenase (LDH), and malondialdehyde (MDA), and the reduction of cell viability, superoxide dismutase (SOD) activity, cytochrome P450 1a (CYP1A), and cytochrome P450 3a (CYP3A) messenger RNA (mRNA) levels expect LDH in the post-treatment. The levels of GPT, GOT, and CYP1A mRNA were also effectively restored in the pre-treatment with phyllanthin (10 μg/ml). Overall, our results suggested that phyllanthin may be used as a hepatoprotective agent to prevent liver diseases in fish.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7

Similar content being viewed by others

References

  • Adeneye AA, Benebo AS (2008) Protective effect of the aqueous leaf and seed extract of Phyllanthus amaru on gentamicin and acetaminophen-induced nephrotoxic rats. J Ethnopharmacol 118:318–323

    Article  PubMed  Google Scholar 

  • 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 41:1–15

    Article  Google Scholar 

  • Allis JW, Brown BL, Simmons JE, Hatch GE, McDonald A, House DE (1996) Methanol potentiation of carbon tetrachloride hepatotoxicity: the central role of cytochrome P450. Toxicology 112:131–140

    Article  PubMed  CAS  Google Scholar 

  • Bainy AC, Saito E, Carvalho PS, Junqueira VB (1996) Oxidative stress in gill, erythrocytes, liver and kidney of Nile tilapia (Oreochromis niloticus) from a polluted site. Aquat Toxicol 34:151–162

    Article  CAS  Google Scholar 

  • Bastien MC, Leblond F, Pichette V, Villeneuve JP (2000) Differential alteration of cytochrome P450 isoenzymes in two experimental models of cirrhosis. Can J Physiol Pharmacol 78:912–919

    Article  PubMed  CAS  Google Scholar 

  • Berry MN, Halls HJ, Grivell MB (1992) Techniques for pharmacological and toxicological studies with isolated hepatocyte suspensions. Life Sci 51:1–16

    Article  PubMed  CAS  Google Scholar 

  • Bhadauria M, Nirala SK, Shrivastava S, Sharma A, Johri S, Chandan BK, Singh B, Saxena AK, Shukla S (2009) Emodin reverses CCl4 induced hepatic cytochrome P450 (CYP) enzymatic and ultrastructural changes: the in vivo evidence. Hepatol Res 39:290–300

    Article  PubMed  CAS  Google Scholar 

  • Casillas E, Myers M, Ames WE (1983) Relationship of serum chemistry values to liver and kidney histopathology in English sole (Parophrys vetulus) after acute exposure to carbon tetrachloride. Aquat Toxicol 3:61–78

    Article  CAS  Google Scholar 

  • Chandan B, Saxena A, Shukla S, Sharma N, Gupta D, Suri K, Suri J, Bhadauria M, Singh B (2007) Hepatoprotective potential of Aloe barbadensis Mill. against carbon tetrachloride induced hepatotoxicity. J Ethnopharmacol 111:560–566

    Article  PubMed  CAS  Google Scholar 

  • 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 261:271–279

    Article  PubMed  CAS  Google Scholar 

  • Chirdchupunseree H, Pramyothin P (2010) Protective activity of phyllanthin in ethanol-treated primary culture of rat hepatocytes. J Ethnopharmacol 128:172–176

    Article  PubMed  CAS  Google Scholar 

  • Chudapongse N, Kamkhunthod M, Poompachee K (2010) Effects of < i > Phyllanthus urinaria</i > extract on HepG2 cell viability and oxidative phosphorylation by isolated rat liver mitochondria. J Ethnopharmacol 130:315–319

    Article  PubMed  Google Scholar 

  • Ding L, Yongqing M, Fasheng C (1990) Nutritional lipid liver disease of grass carp Ctenopharyngodon idullus (C. et V.). Chin J Oceanol Limnol 8:363–373

    Article  Google Scholar 

  • George J, Murray M, Byth K, Farrell GC (1995) Differential alterations of cytochrome P450 proteins in livers from patients with severe chronic liver disease. Hepatology 21:120–128

    PubMed  CAS  Google Scholar 

  • Jia R, Cao L, Xu P, Jeney G, Yin G (2012) In vitro and in vivo hepatoprotective and antioxidant effects of Astragalus polysaccharides against carbon tetrachloride-induced hepatocyte damage in common carp (Cyprinus carpio). Fish Physiol Biochem 38:871–881

    Article  PubMed  CAS  Google Scholar 

  • 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 Deve Biol-Animal:1–7

  • 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 10:87–97

    Article  Google Scholar 

  • Kavitha P, Ramesh R, Bupesh G, Stalin A, Subramanian P (2011) Hepatoprotective activity of Tribulus terrestris extract against acetaminophen-induced toxicity in a freshwater fish (Oreochromis mossambicus). In Vitro Cell Deve Biol-Animal 47:698–706

    Article  CAS  Google Scholar 

  • Kepekçi RA, Polat S, Çelik A, Bayat N, Saygideger SD (2013) Protective effect of Spirulina platensis enriched in phenolic compounds against hepatotoxicity induced by CCl 4. Food Chemistry

  • 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 (< i > Oncorhynchus mykiss</i>). Aquat Toxicol 74:70–81

    Article  PubMed  CAS  Google Scholar 

  • Krithika R, Mohankumar R, Verma RJ, Shrivastav PS, Mohamad IL, Gunasekaran P, Narasimhan S (2009) Isolation, characterization and antioxidative effect of phyllanthin against CCl < sub > 4</sub > −induced toxicity in HepG2 cell line. Chem Biol Interact 181:351–358

    Article  PubMed  CAS  Google Scholar 

  • 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 Experiment Hepatol 1:57–67

    Article  CAS  Google Scholar 

  • Kumar KH, Kuttan R (2004) Protective effect of an extract of Phyllanthus amarus against radiation-induced damage in mice. J Radiat Res 45:133–139

    Article  PubMed  Google Scholar 

  • Lacroix D, Sonnier M, Moncion A, Cheron G, Cresteil T (1997) Expression of CYP3A in the human liver—evidence that the shift between CYP3A7 and CYP3A4 occurs immediately after birth. Eur J Biochem 247:625–634

    Article  PubMed  CAS  Google Scholar 

  • 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 34:471–482

    Article  PubMed  Google Scholar 

  • Lim Y, Murtijaya J (2007) Antioxidant properties of Phyllanthus amarus extracts as affected by different drying methods. LWT-Food Sci Technol 40:1664–1669

    Article  CAS  Google Scholar 

  • Liu X, Zhao M, Wu K, Chai X, Yu H, Tao Z, Wang J (2012) Immunomodulatory and anticancer activities of phenolics from emblica fruit (Phyllanthus emblica L.). Food Chem 131:685–690

    Article  CAS  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT Method. Methods 25:402–408

    Article  PubMed  CAS  Google Scholar 

  • 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). Veterinary Research Forum. pp. 158–164

  • Malhi H, Gores GJ (2008) Cellular and molecular mechanisms of liver injury. Gastroenterology 134:1641–1654

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Manjrekar A, Jisha V, Bag P, Adhikary B, Pai M, Hegde A, Nandini M (2008) Effect of Phyllanthus niruri Linn. treatment on liver, kidney and tests in CCl4 induced hepatotoxic rats. Indian J Exp Biol 46:514

    PubMed  CAS  Google Scholar 

  • Mitra S, Venkataranganna M, Sundaram R, Gopumadhavan S (1998) Effect of HD‐03, a herbal formulation, on the antioxidant defence system in rats. Phytother Res 12:114–117

    Article  Google Scholar 

  • Naaz F, Javed S, Abdin M (2007) Hepatoprotective effect of ethanolic extract of Phyllanthus amarus Schum. et Thonn. on aflatoxin B 1-induced liver damage in mice. J Ethnopharmacol 113:503–509

    Article  PubMed  Google Scholar 

  • Ohnuma T, Anan E, Hoashi R, Takeda Y, Nishiyama T, Ogura K, Hiratsuka A (2011) Dietary diacetylene falcarindiol induces phase 2 drug-metabolizing enzymes and blocks carbon tetrachloride-induced hepatotoxicity in mice through suppression of lipid peroxidation. Biol Pharm Bull 34:371–378

    Article  PubMed  CAS  Google Scholar 

  • 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 50:2310–2317

    Article  CAS  Google Scholar 

  • Passeri MJ, Cinaroglu A, Gao C, Sadler KC (2009) Hepatic steatosis in response to acute alcohol exposure in zebrafish requires sterol regulatory element binding protein activation. Hepatology 49:443–452

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Pikarsky E, Ronen A, Abramowitz J, Levavi-Sivan B, Hutoran M, Shapira Y, Steinitz M, Perelberg A, Soffer D, Kotler M (2004) Pathogenesis of acute viral disease induced in fish by carp interstitial nephritis and gill necrosis virus. J Virol 78:9544–9551

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Råbergh CM, Lipsky MM (1997) Toxicity of chloroform and carbon tetrachloride in primary cultures of rainbow trout hepatocytes. Aquat Toxicol 37:169–182

    Article  Google Scholar 

  • Rajeshkumar N, Joy K, Kuttan G, Ramsewak R, Nair MG, Kuttan R (2002) Antitumour and anticarcinogenic activity of Phyllanthus amarus extract. J Ethnopharmacol 81:17–22

    Article  PubMed  CAS  Google Scholar 

  • Raphael KR, Kuttan R (2003) Inhibition of experimental gastric lesion and inflammation by Phyllanthus amarus extract. J Ethnopharmacol 87:193–197

    Article  PubMed  Google Scholar 

  • Recknagel RO, Glende EA, Dolak JA, Waller RL (1989) Mechanisms of carbon tetrachloride toxicity. Pharmacol Ther 43:139–154

    Article  PubMed  CAS  Google Scholar 

  • Riordan SM, Skinner N, Nagree A, McCallum H, McIver CJ, Kurtovic J, Hamilton JA, Bengmark S, Williams R, Visvanathan K (2003) Peripheral blood mononuclear cell expression of toll‐like receptors and relation to cytokine levels in cirrhosis. Hepatology 37:1154–1164

    Article  PubMed  CAS  Google Scholar 

  • Rodriguez-Antona C, Donato MT, Pareja E, Gomez-Lechon M-J, Castell JV (2001) Cytochrome P-450 mRNA expression in human liver and its relationship with enzyme activity. Arch Biochem Biophys 393:308–315

    Article  PubMed  CAS  Google Scholar 

  • 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 252:165–175

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Shen X, Tang Y, Yang R, Yu L, Fang T, J-a D (2009) The protective effect of Zizyphus jujube fruit on carbon tetrachloride-induced hepatic injury in mice by anti-oxidative activities. J Ethnopharmacol 122:555–560

    Article  PubMed  CAS  Google Scholar 

  • Shim J-Y, Kim M-H, Kim H-D, Ahn J-Y, Yun Y-S, Song J-Y (2010) Protective action of the immunomodulator ginsan against carbon tetrachloride-induced liver injury via control of oxidative stress and the inflammatory response. Toxicol Appl Pharmacol 242:318–325

    Article  PubMed  CAS  Google Scholar 

  • Shivashri C, Rajarajeshwari T, Rajasekar P (2013) Hepatoprotective action of celery (Apium graveolens) leaves in acetaminophen-fed freshwater fish (Pangasius sutchi). Fish Physiol Biochem:1–13

  • Sugiyama T, Nagata J-i, Yamagishi A, Endoh K, Saito M, Yamada K, Yamada S, Umegaki K (2006) Selective protection of curcumin against carbon tetrachloride-induced inactivation of hepatic cytochrome P450 isozymes in rats. Life Sci 78:2188–2193

    Article  PubMed  CAS  Google Scholar 

  • Taesotikul T, Dumrongsakulchai W, Wattanachai N, Navinpipat V, Somanabandhu A, Tassaneeyakul W, Tassaneeyakul W (2010) Inhibitory effects of Phyllanthus amarus and its major lignans on human microsomal cytochrome P450 activities: evidence for CYP3A4 mechanism-based inhibition. Drug Metab Pharmacokinet 41:1012–1018

    Google Scholar 

  • Thibaut R, Schnell S, Porte C (2006) The interference of pharmaceuticals with endogenous and xenobiotic metabolizing enzymes in carp liver: an in-vitro study. Environment Sci Technol 40:5154–5160

    Article  CAS  Google Scholar 

  • Venkatesan P, Satyan K, Sudheer Kumar M, Prakash A (2003) Protective effect by aqueous extract of Phyllanthus amarus linn., phyllanthin and nirocil against carbontetrachloride-induced liver and brain toxicity. Ind J Pharmaceut Sci 65:309–312

    Google Scholar 

  • Wang L-L, Liu T, Wang C, Zhao F-Q, Zhang Z-W, Yao H-D, Xing H-J, Xu S-W (2013) Effects of atrazine and chlorpyrifos on the production of nitric oxide and expression of inducible nitric oxide synthase in the brain of common carp (Cyprinus carpio L.). Ecotoxicol Environ Saf 43:324–239

    Google Scholar 

  • Weber LW, Boll M, Stampfl A (2003) Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. CRC Crit Rev Toxicol 33:105–136

    Article  CAS  Google Scholar 

  • Wolf JC, Wolfe MJ (2005) A brief overview of nonneoplastic hepatic toxicity in fish. Toxicol Pathol 33:75–85

    Article  PubMed  CAS  Google Scholar 

  • Xie Y, Hao H, Kang A, Liang Y, Xie T, Sun S, Dai C, Zheng X, Xie L, Li J (2010) Integral pharmacokinetics of multiple lignan components in normal, CCl < sub > 4</sub > −induced hepatic injury and hepatoprotective agents pretreated rats and correlations with hepatic injury biomarkers. J Ethnopharmacol 131:290–299

    Article  PubMed  CAS  Google Scholar 

  • Yadav NP, Pal A, Shanker K, Bawankule DU, Gupta AK, Darokar MP, Khanuja SPS (2008) Synergistic effect of silymarin and standardized extract of Phyllanthus amarus against CCl 4-induced hepatotoxicity in Rattus norvegicus. Phytomedicine 15:1053–1061

    Article  PubMed  CAS  Google Scholar 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • Yokogawa K, Watanabe M, Takeshita H, Nomura M, Mano Y, Miyamoto K-i (2004) Serum aminotransferase activity as a predictor of clearance of drugs metabolized by CYP isoforms in rats with acute hepatic failure induced by carbon tetrachloride. Int J Pharm 269:479–489

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y, Jia Y, Yang M, Yang P, Tian Y, Xiao A, Wen A (2012) The impaired disposition of probe drugs is due to both liver and kidney dysfunctions in CCl 4-model rats. Environ Toxicol Pharmacol 33:453–458

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by National Natural Science Foundation of China (31202002, 31200918).

Conflict of interest

The authors declare that they have no competing financial or non-financial interests.

Author information

Authors and Affiliations

  1. Wuxi Fisheries College, Nanjing Agricultural University, Wuxi, 214081, China

    Rui Jia, Ying-Juan Liu, Pao Xu & Guo-Jun Yin

  2. Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Science, Wuxi, 214081, China

    Rui Jia, Jin-Liang Du, Li-Ping Cao, Pao Xu & Guo-Jun Yin

  3. International Joint Research Laboratory for Fish Immunopharmacology, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi, 214081, China

    Jin-Liang Du, Li-Ping Cao, Pao Xu & Guo-Jun Yin

Authors
  1. Rui Jia
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jin-Liang Du
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Li-Ping Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ying-Juan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Pao Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Guo-Jun Yin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Guo-Jun Yin.

Additional information

Editor: Tetsuji Okamoto

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Jia, R., Du, JL., Cao, LP. et al. Protective action of the phyllanthin against carbon tetrachloride-induced hepatocyte damage in Cyprinus carpio . In Vitro Cell.Dev.Biol.-Animal 52, 1–9 (2016). https://doi.org/10.1007/s11626-015-9946-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-015-9946-3

Keywords

Search

Navigation