Advertisement

Inflammopharmacology

, Volume 18, Issue 2, pp 87–94 | Cite as

Hepatoprotective effects of the nitric oxide donor isosorbide-5-mononitrate alone and in combination with the natural hepatoprotectant, silymarin, on carbon tetrachloride-induced hepatic injury in rats

  • Omar Mohamed E. Abdel Salam
  • Amany A. Sleem
  • Nermeen Shafee
Research Article

Abstract

The aim of this study was to investigate the effect of the nitric oxide donor isosorbide-5-mononitrate (5-ISMN) alone or in combination with the natural hepatoprotectant with anti-oxidant activity silymarin on the carbon tetrachloride (CCl4)-induced hepatic injury in rats. 5-ISMN (1.8, 3.6 or 7.2 mg/kg), silymarin (25 mg/kg) or 5-ISMN (1.8, 3.6 or 7.2 mg/kg) combined with silymarin was given once daily orally simultaneously with CCl4 and for 15 days thereafter. Liver damage was assessed by determining serum enzyme activities and hepatic histopathology. 5-ISMN given at the above doses conferred significant protection against the hepatotoxic actions of CCl4 in rats, reducing serum alanine aminotransferase (ALT) levels by 31.2, 39.3 and 61.6%, respectively, when compared with controls. Serum aspartate aminotransferase (AST) levels decreased by 19.8, 22.7 and 59.4%, respectively, while alkaline phosphatase (ALP) decreased by 26.1 and 32.6% by the drug at 3.6 and 7.2 mg/kg, respectively. When silymarin was added to 5-ISMN (1.8, 3.6 or 7.2 mg/kg), ALT decreased by 32.8, 59.6, 70.2% and AST by 28.7, 50.3, 60%, when compared with CCl4 control group levels. Silymarin in combination with 3.6 or 7.2 mg/kg 5-ISMN resulted in 37.5 and 39.2% reductions in ALP when compared with CCl4 control group. Meanwhile, silymarin alone reduced ALT, AST and ALP levels by 65.9, 52 and 62.3%, respectively. Blood levels of reduced glutathione were markedly decreased in CCl4-treated rats. Reduced glutathione levels were increased by the administration of 5-ISMN and restored to near normal values by silymarin treatment. Histopathological alterations by CCl4 were markedly reduced after treatment with 5-ISMN alone or in combination with silymarin. Histopathologic examination of the livers of CCl4-treated rats administered 5-ISMN at 7.2 mg/kg showed marked restoration of the normal architecture of the liver tissue and minimal fibrosis. Silymarin co-administered with 5-ISMN resulted in further improvement of the histologic picture. These results indicates that treatment with 5-ISMN protects against hepatocellular necrosis induced by CCl4. The study suggests a potential therapeutic use for 5-ISMN in combination with silymarin in liver injury.

Keywords

Isosorbide-5-mononitrate Silymarin Acute hepatic injury Carbon tetrachloride Rat 

References

  1. Abdel Salam OME, Sleem AA, Omara EA et al (2007) Effect of ribavirin alone or combined with silymarin on carbon tetrachloride induced hepatic damage in rats. Drug Target Insights 1:19–27Google Scholar
  2. Abdel Salam OME, Sleem AA, Omara EA et al (2009) Hepatoprotective effects of misoprostol and silymarin on carbon tetrachloride-induced hepatic damage in rats. Fundam Clin Pharmacol 23:179–188CrossRefGoogle Scholar
  3. Abrams J (1989) Long-acting nitrates in cardiovascular disease. In: Giles TD, Garbus SB (eds) Use of longer-acting agents in cardiac therapy. Recent advances. Health Care Communications, Inc, USA, pp 107–128Google Scholar
  4. Abshagen UWP (1992) Pharmacokinetics of isosorbide mononitrate. Am J Cardiol 70:61G–66GCrossRefPubMedGoogle Scholar
  5. Alter M, Kruszon-Moran D, Nainan O et al (1994) The prevalence of hepatitis C virus infection in the United States, 1988 through 1994. N Engl J Med 341:556–562CrossRefGoogle Scholar
  6. Bani D, Nistri S, Quattrone S et al (2001) The vasorelaxant hormone relaxin induces changes in liver sinusoid microcirculation: a morphologic study in the rat. J Endocrinol 171:541–549CrossRefPubMedGoogle Scholar
  7. Belfield A, Goldberg DM (1971) Revised assay for serum phenyl phosphatase activity using 4-amino-antipyrine. Enzyme 12:561–573PubMedGoogle Scholar
  8. Beutler E, Duron O, Kelly BM (1963) Improved method for the determination of blood glutathione. J Lab Clin Med 61:882–890PubMedGoogle Scholar
  9. Boigk G, Stroedter L, Herbst H et al (1997) Silymarin retards collagen accumulation in early and advanced biliary fibrosis secondary to complete bile duct obliteration in rats. Hepatology 26:643–649CrossRefPubMedGoogle Scholar
  10. Burra P, Targhetta S, Pevere S et al (2006) Antiviral therapy for hepatitis C virus recurrence following liver transplantation: long-term results from a single center experience. Transplant Proc 38:1127–1130CrossRefPubMedGoogle Scholar
  11. Chazouillères O, Ballet F, Chrétien Y et al (1989) Protective effect of vasodilators on liver function after long hypothermic preservation: a study in the isolated perfused rat liver. Hepatology 9:824–829CrossRefPubMedGoogle Scholar
  12. Cnubben NHP, Rietjens IMCM, Wortelboer H et al (2000) The interplay of glutathione-related processes in antioxidant defense. Environ Toxicol Pharmacol 10:141–152CrossRefGoogle Scholar
  13. Crowley LV (1967) The Reitman-Frankel colorimetric transaminase procedure in suspected myocardial infarction. Clin Chem 13:482–487PubMedGoogle Scholar
  14. De La Puerta R, Martinez E, Bravo L et al (1996) Effect of silymarin on different acute inflammation models and on leukocyte migration. J Pharm Pharmacol 48:968–970PubMedGoogle Scholar
  15. Dehmlow C, Murawski N, De Groot H (1996) Scavenging of reactive oxygen species and inhibition of arachidonic acid metabolism by silibinin in human cells. Life Sci 58:1591–1600CrossRefPubMedGoogle Scholar
  16. Failli P, DeFranco RM, Caligiuri AM et al (2000) Nitrovasodilators inhibit platelet-derived growth factor-induced proliferation and migration of activated human hepatic stellate cells. Gastroenterology 119:479–492CrossRefPubMedGoogle Scholar
  17. Feher J, Deak G, Muzes G et al (1989) Liver protective action of silymarin therapy in chronic alcoholic liver diseases. Orv Hetil 130:2723–2727PubMedGoogle Scholar
  18. Feher J, Lengyel G, Blazovics A (1998) Oxidative stress in the liver and biliary tract diseases. Scand J Gastroenterol Suppl 228:38–46PubMedGoogle Scholar
  19. Fiorucci S, Antonelli E, Distrutti E et al (2004) Liver delivery of NO by NCX-1000 protects against acute liver failure and mitochondrial dysfunction induced by APAP in mice. Br J Pharmacol 143:33–42CrossRefPubMedGoogle Scholar
  20. Flora K, Hahn M, Rosen H et al (1998) Milk thistle (Silybum marianum) for the therapy of liver disease. Am J Gastroenterol 93:139–143CrossRefPubMedGoogle Scholar
  21. Förstermann U, Pollock JS, Schmidt HH et al (1991) Calmodulin-dependent endothelium-derived relaxing factor/nitric oxide synthase activity is present in the particulate and cytosolic fractions of bovine aortic endothelial cells. Proc Natl Acad Sci USA 88:1788–1792CrossRefPubMedGoogle Scholar
  22. Fraschini F, Demartini G, Esposti D (2002) Pharmacology of silymarin. Clin Drug Invest 22:51–65CrossRefGoogle Scholar
  23. Fried MW, Shiffman ML, Reddy KR et al (2002) Peginterferon alfa-2a plus ribavirin for chronic hepatitis C virus infection. N Engl J Med 347:975–982CrossRefPubMedGoogle Scholar
  24. García-Pagán JC, Navasa M, Bosch J et al (1990) Enhancement of portal pressure reduction by the association of isosorbide-5-mononitrate to propranolol administration in patients with cirrhosis. Hepatology 11:230–238CrossRefPubMedGoogle Scholar
  25. García-Pagán JC, Feu F, Bosch J et al (1991) Propranolol compared with propranolol plus isosorbide-5-mononitrate for portal hypertension in cirrhosis. A randomized controlled study. Ann Intern Med 114:869–873PubMedGoogle Scholar
  26. Gordon A, Hobbs DA, Bowden DS et al (2006) Effects of Silybum marianum on serum hepatitis C virus RNA, alanine aminotransferase levels and well-being in patients with chronic hepatitis C. J Gastroenterol Hepatol 21:275–280CrossRefPubMedGoogle Scholar
  27. Hayes PC, Westaby D, Williams R (1998) Effect and mechanism of action of isosorbide-5-mononitrate. Gut 29:752–755CrossRefGoogle Scholar
  28. Hoofnagle JH, Ghany MG, Kleiner DE et al (2003) Maintenance therapy with ribavirin in patients with chronic hepatitis C who fail to respond to combination therapy with interferon alfa and ribavirin. Hepatology 38:66–74CrossRefPubMedGoogle Scholar
  29. Hutt V, Theodor R, Pabst G et al (1992) Evaluation of bioavailability and pharmacokinetics of two isosorbide-5-mononitrate preparations in healthy volunteers. J Clin Pharmacol 32:553–557CrossRefPubMedGoogle Scholar
  30. Ijaz S, Yang W, Winslet MC et al (2005) The role of nitric oxide in the modulation of hepatic microcirculation and tissue oxygenation in an experimental model of hepatic steatosis. Microvasc Res 70:129–136CrossRefPubMedGoogle Scholar
  31. Jia JD, Bauer M, Cho JJ et al (2001) Antifibrotic effect of silymarin in rat secondary biliary fibrosis is mediated by downregulation of procollagen alpha1(I) and TIMP-1. J Hepatol 35:392–398CrossRefPubMedGoogle Scholar
  32. Khodosovskii MN, Zinchuk VV (2006) Effect of nitroglycerin on some parameters of the prooxidant-antioxidant balance and functional state of the liver during ischemia/reperfusion. Bull Exp Biol Med 142:675–678CrossRefPubMedGoogle Scholar
  33. Kim WR, Brown RS Jr, Terrault NA et al (2002) Burden of liver disease in the United States: summary of a workshop. Hepatology 36:227–242CrossRefPubMedGoogle Scholar
  34. Knowles RG, Moncada S (1994) Nitric oxide synthases in mammals. Biochem J 298:249–258PubMedGoogle Scholar
  35. Kosaka H, Seiyama A (1997) Elevation of oxygen release by nitroglycerin without an increase in blood flow in the hepatic microcirculation. Nat Med 3:456–459CrossRefPubMedGoogle Scholar
  36. Lang JD Jr, Teng X, Chumley P et al (2007) Inhaled NO accelerates restoration of liver function in adults following orthotopic liver transplantation. J Clin Invest 117:2583–2591CrossRefPubMedGoogle Scholar
  37. Lhuillier F, Robert MO, Crova P et al (2006) Nitric oxide and liver microcirculation during autoregulation and haemorrhagic shock in rabbit model. Br J Anaesth 97:137–146CrossRefPubMedGoogle Scholar
  38. Li X, Benjamin IS, Alexander B (2003) The role of nitric oxide in systemic and hepatic haemodynamics in the rat in vivo. Naunyn Schmiedebergs Arch Pharmacol 368:142–149CrossRefPubMedGoogle Scholar
  39. Lieber CS, Leo MA, Cao Q et al (2003) Silymarin retards the progression of alcohol-induced hepatic fibrosis in baboons. J Clin Gastroenterol 37:336–339CrossRefPubMedGoogle Scholar
  40. Loureiro-Silva MR, Cadelina GW, Iwakiri Y et al (2003) A liver-specific nitric oxide donor improves the intra-hepatic vascular response to both portal blood flow increase and methoxamine in cirrhotic rats. J Hepatol 39:940–946CrossRefPubMedGoogle Scholar
  41. Lucena MI, Andrade RJ, de la Cruz JP et al (2002) Effects of silymarin MZ-80 on oxidative stress in patients with alcoholic cirrhosis. Results of a randomized, double-blind, placebo-controlled clinical study. Int J Clin Pharmacol Ther 40:2–8PubMedGoogle Scholar
  42. Mayer KÉ, Myers RP, Lee SS (2005) Silymarin treatment of viral hepatitis: a systematic review. J Viral Hepatitis 12:559–567CrossRefGoogle Scholar
  43. Merkel C, Sacerdoti C, Bolognesi M et al (1997) Hemodynamic evaluation of the addition of isosorbide-5-mononitrate to nadolol in cirrhotic patients with insufficient response to the beta-blocker alone. Hepatology 26:34–39PubMedGoogle Scholar
  44. Mourelle M, Muriel P, Favari L et al (1989) Prevention of CCl4-induced liver cirrhosis by silymarin. Fundam Clin Pharmacol 3:183–191CrossRefPubMedGoogle Scholar
  45. Mukherjee S, Lyden E (2006) Impact of pegylated interferon alpha-2B and ribavirin on hepatic fibrosis in liver transplant patients with recurrent hepatitis C: an open-label series. 1. Liver Int 26:529–535CrossRefPubMedGoogle Scholar
  46. Muriel P (1998) Nitric oxide protection of rat liver from lipid peroxidation, collagen accumulation, and liver damage induced by carbon tetrachloride. Biochem Pharmacol 56:773–779CrossRefPubMedGoogle Scholar
  47. Muriel P, Mourelle M (1990) Prevention by silymarin of membrane alterations in acute CCl4 liver damage. J Appl Toxicol 10:275–279CrossRefPubMedGoogle Scholar
  48. Muriel P, Garciapina T, Perez-Alvarez V et al (1992) Silymarin protects against paracetamol-induced lipid peroxidation and liver damage. J Appl Toxicol 12:439–442CrossRefPubMedGoogle Scholar
  49. Navasa M, Chesta J, Bosch J et al (1989) Reduction of portal pressure by isosorbide-5-mononitrate in patients with cirrhosis. Effects on splanchnic and systemic hemodynamics and liver function. Gastroenterology 96:1110–1118PubMedGoogle Scholar
  50. Ohata K, Hamasaki K, Toriyama K et al (2004) High viral load is a risk factor for hepatocellular carcinoma in patients with chronic hepatitis B virus infection. J Gastroenterol Hepatol 19:670–675CrossRefPubMedGoogle Scholar
  51. Paget GE, Barnes JM (1964) Toxicity testing. In: Laurence DR, Bacharach AL (eds) Evaluation of drug activities pharmacometics. Academic Press, London, pp 1–135Google Scholar
  52. Pár A, Roth E, Rumi G et al (2000) Oxidative stress and antioxidant defense in alcoholic liver disease and chronic hepatitis C. Orv Hetil 141:1655–1659PubMedGoogle Scholar
  53. Pares A, Planas R, Torres M et al (1998) Effects of silymarin in alcoholic patients with cirrhosis of the liver: results of a controlled, double-blind, randomized and multicenter trial. J Hepatol 28:615–621CrossRefPubMedGoogle Scholar
  54. Parker JD, Parker JO (1998) Nitrate therapy for stable angina pectoris. N Engl J Med 338:520–531CrossRefPubMedGoogle Scholar
  55. Pianko S, McHutchison JG (2000) Treatment of hepatitis C with interferon and ribavirin. J Gastroenterol Hepatol 15:581–586CrossRefPubMedGoogle Scholar
  56. Saller R, Meier R, Brignoli R (2001) The use of silymarin in the treatment of liver diseases. Drugs 61:2035–2063CrossRefPubMedGoogle Scholar
  57. Schneider W, Menke G, Heidemann R et al (1990) Concentrations of isosorbide dinitrate, isosorbide-2-mononitrate and isosorbide-5-mononitrate in human vascular and muscle tissue under steady-state conditions. Eur J Clin Pharmacol 38:145–147CrossRefPubMedGoogle Scholar
  58. Slehria S, Rajvanshi P, Ito Y, Sokhi RP et al (2002) Hepatic sinusoidal vasodilators improve transplanted cell engraftment and ameliorate microcirculatory perturbations in the liver. Hepatology 35:1320–1328CrossRefPubMedGoogle Scholar
  59. Stanley AJ, Bouchier IAD, Hayes PC (1998) Acute effect of propranolol and isosorbide-5-mononitrate administration on renal blood flow in cirrhotic patients. Gut 42:283–287PubMedCrossRefGoogle Scholar
  60. Wellington K, Jarvis B (2001) Silymarin: a review of its clinical properties in the management of hepatic disorders. BioDrugs 15:465–489CrossRefPubMedGoogle Scholar
  61. Wu CG, Chamuleau RA, Bosch KS et al (1993) Protective effect of silymarin on rat liver injury induced by ischemia. Virchows Arch B Cell Pathol 64:259–263CrossRefGoogle Scholar
  62. Zhang WH, Li YJ, Zhou Y (2006) Melatonin abates liver ischemia/reperfusion injury by improving the balance between nitric oxide and endothelin. Hepatobiliary Pancreat Dis Int 5:574–579PubMedGoogle Scholar

Copyright information

© Birkhäuser Verlag, Basel/Switzerland 2010

Authors and Affiliations

  • Omar Mohamed E. Abdel Salam
    • 1
  • Amany A. Sleem
    • 1
  • Nermeen Shafee
    • 2
  1. 1.Department of PharmacologyNational Research CentreCairoEgypt
  2. 2.Department of PathologyNational Research CentreCairoEgypt

Personalised recommendations