Inflammation Research

, Volume 58, Issue 9, pp 593–599

Hepatic protection by glycyrrhizin and inhibition of iNOS expression in concanavalin A-induced liver injury in mice

  • Noriko Tsuruoka
  • Kazuki Abe
  • Kenjirou Wake
  • Masaru Takata
  • Akira Hatta
  • Tositugu Sato
  • Hideo Inoue
Original Research Paper

Abstract

Objective and design

In this study, the possible protective effect of glycyrrhizin (GL), an active compound derived from licorice root, was examined on T cell-mediated liver injury in mice.

Materials and methods

Mice were subjected to liver injury by intravenous injection of concanavalin A (Con A). They had been treated with GL (i.p.) 30 min before the injection. Liver injury was estimated by measuring serum levels of alanine aminotransaminase (ALT) and aspartate aminotransaminase (AST), and by examining liver sections with hematoxylin–eosin staining. Expression of inducible nitric oxide synthase (iNOS) mRNA and protein in the liver was determined by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting.

Results

Serum transaminases and hepatic iNOS levels increased with time after Con A treatment. Expression of iNOS mRNA in the liver was elevated for up to 8 h, and at 8 h, GL (ED50: 10.5 mg/kg) suppressed the increases in AST and ALT in response to Con A. An increase in iNOS mRNA expression and protein was inhibited by treatment with GL. Furthermore, GL inhibited cell infiltration and the degeneration of hepatocytes in the liver of Con A-treated mice.

Conclusion

The present study suggests that the prevention by GL of Con A-induced hepatitis is due partly to the modulation of hepatic iNOS induction and of degeneration of hepatocytes.

Keywords

Glycyrrhizin Concanavalin A-liver injury Inducible nitric oxide synthase Hepatic protection 

References

  1. 1.
    Yoshikawa M, Matsui Y, Kawamoto H, Umemoto N, Oku K, Koizumi M, et al. Effects of glycyrrhizin on immune-mediated cytotoxicity. J Gastroenterol Hepatol. 1997;12:243–8.PubMedCrossRefGoogle Scholar
  2. 2.
    Okamoto T, Kanda T. Glycyrrhizin protects mice from concanavalin A-induced hepatitis without affecting cytokine expression. Int J Mol Med. 1999;4:149–52.PubMedGoogle Scholar
  3. 3.
    Okamoto T. The protective effect of glycyrrhizin on anti-Fas antibody-induced hepatitis in mice. Eur J Pharmacol. 2000;387:229–32.PubMedCrossRefGoogle Scholar
  4. 4.
    Kimura M, Inoue H, Hirabayashi K, Natsume H, Ogihara M. Glycyrrhizin and some analogues induce growth of primary cultured adult rat hepatocytes via epidermal growth factor receptors. Eur J Pharmacol. 2001;431:151–61.PubMedCrossRefGoogle Scholar
  5. 5.
    Kimura M, Moro T, Motegi H, Maruyama H, Sekine M, Okamoto H, et al. In vivo glycyrrhizin accelerates liver regeneration and rapidly lowers serum transaminase activities in 70% partially hepatectomized rats. Eur J Pharmacol. 2008;579:357–64.PubMedCrossRefGoogle Scholar
  6. 6.
    Yin J, Li D, Hu W, Meng Q. Effects of glycyrrhizic acid on cocklebur-induced hepatotoxicity in rat and human hepatocytes. Phytother Res. 2008;22:395–400.PubMedCrossRefGoogle Scholar
  7. 7.
    Tang B, Qiao H, Meng F, Sun X. Glycyrrhizin attenuates endotoxin-induced acute liver injury after partial hepatectomy in rats. Braz J Med Biol Res. 2007;40:1637–46.PubMedCrossRefGoogle Scholar
  8. 8.
    Yoshida T, Abe K, Ikeda T, Matsushita T, Wake K, Sato T, et al. Inhibitory effect of glycyrrhizin on lipopolysaccharide and d-galactosamine-induced mouse liver injury. Eur J Pharmacol. 2007;576:136–42.PubMedCrossRefGoogle Scholar
  9. 9.
    Abe K, Ikeda T, Wake K, Sato T, Sato T, Inoue H. Glycyrrhizin privents of lipopolysaccharide/d-galactosamine-induced liver injury through down-regulation of matrix metalloproteinase-9 in mice. J Pharm Pharmacol. 2008;60:91–7.PubMedCrossRefGoogle Scholar
  10. 10.
    Iino S, Tango T, Matsushima T, Toda G, Miyake K, Hino K, et al. Therapeutic effects of stronger neo-minophagen C at different doses on chronic hepatitis and liver cirrhosis. Hepatol Res. 2001;19:31–40.PubMedCrossRefGoogle Scholar
  11. 11.
    Arase Y, Ikeda K, Murashima N, Chayama K, Tsubota A, Koida I, et al. The long term efficacy of glycyrrhizin in chronic hepatitis C patients. Cancer. 1997;79:1494–500.PubMedCrossRefGoogle Scholar
  12. 12.
    Ikeda M, Fujiyama S, Tanaka M, Sata M, Ide T, Yatsuhashi H, et al. Clinical features of hepatocellular carcinoma that occur after sustained virological response to interferon for chronic hepatitis C. J Gastroenterol Hepatol. 2006;21:122–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Hidaka I, Hino K, Korenaga M, Gondo T, Nishina S, Ando M, et al. Stronger Neo-Minophagen C, a glycyrrhizin-containing preparation, protects liver against carbon tetrachloride-induced oxidative stress in transgenic mice expressing the hepatitis C virus polyprotein. Liver Int. 2007;27:845–53.PubMedCrossRefGoogle Scholar
  14. 14.
    van Rossum TG, Vulto AG, Hop WC, Schalm SW. Pharmacokinetics of intravenous glycyrrhizin after single and multiple doses in patients with chronic hepatitis C infection. Clin Ther. 1999;21:2080–90.PubMedCrossRefGoogle Scholar
  15. 15.
    van Rossum TG, Vulto AG, Hop WC, Schalm SW. Glycyrrhizin-induced reduction of ALT in European patients with chronic hepatitis C. Am J Gastroenterol. 2001;96:2432–7.PubMedCrossRefGoogle Scholar
  16. 16.
    Tiegs G, Hentschel J, Wendel A. A T cell-dependent experimental liver injury in mice inducible by concanavalin A. J Clin Invest. 1992;90:196–203.PubMedCrossRefGoogle Scholar
  17. 17.
    Sass G, Heinlein S, Agli A, Bang R, Schümann J, Tiegs G. Cytokine expression in three mouse models of experimental hepatitis. Cytokine. 2002;19:115–20.PubMedCrossRefGoogle Scholar
  18. 18.
    Jaruga B, Hong F, Kim WH, Gao B. IFN-gamma/STAT1 acts as a proinflammatory signal in T cell-mediated hepatitis via induction of multiple chemokines and adhesion molecules: a critical role of IRF-1. Am J Physiol Gastrointest Liver Physiol. 2004;287:G1044–52.PubMedCrossRefGoogle Scholar
  19. 19.
    Fujikura S, Mizuhara H, Miyazawa Y, Fujiwara H, Kaneda K. Kinetics and localization of lymphoblasts that proliferate in the mutine liver after Concanavalin A administration. Biomed Res. 1996;17:129–39.Google Scholar
  20. 20.
    Beckman JS, Koppenol WH. Nitric oxide, superoxide, and peroxynitrite: the good, the bad, and ugly. Am J Physiol. 1996;271:C1424–37.PubMedGoogle Scholar
  21. 21.
    Halliwell B. What nitrates tyrosine? Is nitrotyrosine specific as a biomarker of peroxynitrite formation in vivo? FEBS Lett. 1997;411:157–60.PubMedCrossRefGoogle Scholar
  22. 22.
    Christopherson KS, Bredt DS. Nitric oxide in excitable tissues: physiological roles and diseases. J Clin Invest. 1997;100:2424–9.PubMedCrossRefGoogle Scholar
  23. 23.
    Harrison DG. Cellular and molecular mechanisms of endothelial cell dysfunction. J Clin Invest. 1997;100:2153–7.PubMedCrossRefGoogle Scholar
  24. 24.
    Sass G, Koerber K, Bang R, Guehring H, Tiegs G. Inducible nitric oxide synthase is critical for immune-mediated liver injury in mice. J Clin Invest. 2001;107:439–47.PubMedCrossRefGoogle Scholar
  25. 25.
    Ito Y, Abril Edward R, Bethea Nancy W, McCuskey Robert S. Role of nitric oxide in hepatic microvascular injury elicited by acetaminophen in mice. Am J Physiol Gastrointest Liver Physiol. 2004;286:G60–7.PubMedCrossRefGoogle Scholar
  26. 26.
    Ma XL, Li YH, Gao JX, Li J, Guo L, Wu CZ. Expression of inducible nitric oxide synthase in the liver is under the control of nuclear factor kappa B in concanavalin A-induced hepatitis. J Gastroenterol Hepatol. 2008;23:e231–5.PubMedCrossRefGoogle Scholar
  27. 27.
    Ohuchi K, Kamada Y, Levine L, Tsurufuji S. Glycyrrhizin inhibits prostaglandin E2 production by activated peritoneal macrophages from rats. Prostaglandins Med. 1981;7:457–63.PubMedCrossRefGoogle Scholar
  28. 28.
    Okimasu E, Moromizato Y, Watanabe S, Sasaki J, Shiraishi N, Morimoto YM, et al. Inhibition of phospholipase A2 and platelet aggregation by glycyrrhizin, an antiinflammation drug. Acta Med Okayama. 1983;37:385–91.PubMedGoogle Scholar
  29. 29.
    Feng C, Wang H, Yao C, Zhang J, Tian Z. Diammonium glycyrrhizinate, a component of traditional Chinese medicine Gan-Cao, prevents murine T-cell-mediated fulminant hepatitis in IL-10- and IL-6-dependent manners. Int Immunopharmacol. 2007;7:1292–8.PubMedCrossRefGoogle Scholar
  30. 30.
    Abe M, Akbar F, Hasebe A, Horiike N, Onji M. Glycyrrhizin enhances interleukin-10 production by liver dendritic cells in mice with hepatitis. J Gastroenterol. 2003;38:962–7.PubMedCrossRefGoogle Scholar
  31. 31.
    Lee CH, Park SW, Kim YS, Kang SS, Kim JA, Lee SH, et al. Protective mechanism of glycyrrhizin on acute liver injury induced by carbon tetrachloride in mice. Biol Pharm Bull. 2007;30:1898–904.PubMedCrossRefGoogle Scholar
  32. 32.
    Wang JY, Guo JS, Li H, Liu SL, Zern MA. Inhibitory effect of glycyrrhizin on NF-kappaB binding activity in CCl4- plus ethanol-induced liver cirrhosis in rats. Liver. 1998;18:180–5.PubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag, Basel/Switzerland 2009

Authors and Affiliations

  • Noriko Tsuruoka
    • 1
  • Kazuki Abe
    • 1
  • Kenjirou Wake
    • 2
  • Masaru Takata
    • 1
  • Akira Hatta
    • 1
  • Tositugu Sato
    • 1
  • Hideo Inoue
    • 1
  1. 1.Pharmacological Research DepartmentMinophagen Pharmaceutical Co.ZamaJapan
  2. 2.Liver Research UnitMinophagen Pharmaceutical Co.TokyoJapan

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