Advertisement

Digestive Diseases and Sciences

, Volume 52, Issue 12, pp 3348–3354 | Cite as

The Effects of N-Acetylcysteine on Bile Duct Ligation–Induced Liver Fibrosis in Rats

  • Gulgun Tahan
  • Orhan Tarcin
  • Veysel Tahan
  • Fatih Eren
  • Nursal Gedik
  • Elife Sahan
  • Nilgun Biberoglu
  • Savas Guzel
  • Aysun Bozbas
  • Nurdan Tozun
  • Osman Yucel
Original Paper

Abstract

Stellate cells are activated by free radicals, and synthesize collagen. N-acetylcysteine (NAC) is a precursor of reduced glutathione and a potent scavenger of hydroxyl radicals and has potential antifibrotic effects. We aimed to test the effects of NAC on bile duct ligation (BDL) induced liver damage in rats. Forty-seven Wistar rats were divided into 5 groups: group 1, BDL + NAC (n = 10); group 2, BDL (n = 10); group 3, sham + NAC (n = 10); group 4, sham (n = 10); and group 5, control group (n = 10). NAC (50 μmol/kg per day) or saline of single doses were administered intraperitoneally for 28 days. Serum biochemical and liver oxidative stress parameters were studied. Liver collagen level was determined by the method of Lopez de Leon and Rojkind. Liver slides were stained by hematoxylin and eosin and Masson trichrome\Gomory reticulum staining. Aspartate aminotransferase (AST) and alkaline phosphatase levels in the BDL + NAC group were lower than the BDL group and were higher than the control groups (all P < .001). Malondialdehyde, luminal, and glutathione levels in group 1 were lower than the BDL group (P = .01, P = .002, and P < .001) and higher than the control groups (all P < .001). NAC had no effect on alanine aminotransferase (ALT), gammaglutamyl transferase, bilirubin, albumin, or lucigenin levels. Liver collagen levels were higher in the BDL groups (P < .001); however, NAC had no effect on the collagen levels. The BDL groups showed stage 3 fibrosis; all the control groups were normal. NAC improved some biochemical parameters (AST, alkaline phosphatase) and oxidative stress parameters (malondialdehyde, luminol, glutathione) in the BDL model. NAC was found to be effective on cholestasis-induced hepatotoxicity. However, NAC was inefficient as an antifibrotic agent within a 1-month period of administration in the BDL model.

Keywords

Antifibrotic N-acetylcysteine Bile duct ligation Hepatic fibrosis Rat 

References

  1. 1.
    Svegilati Baroni G, D’Ambrosio L, Ferretti G, Casini A, Di Sario A, Salzano R, Ridolfi F, Saccomanno S, Jezequel AM, Benedetti A (1998) Fibrogenic effect of oxidative stress on rat hepatic stellate cells. Hepatology 27:720–726CrossRefGoogle Scholar
  2. 2.
    Parola M, Leonarduzzi G, Robino G, Albano E, Poli G (1996) On the role of lipid peroxidation in the pathogenesis of liver damage induced by long-standing cholestasis. Free Radical Biol Med 20:351–359CrossRefGoogle Scholar
  3. 3.
    Tahan V, Ozaras R, Canbakan B, Uzun H, Aydin S, Yildirim B, Aytekin H, Ozbay G, Mert A, Senturk H (2004) Melatonin reduces dimethylnitrosamine-induced liver fibrosis in rats. J Pineal Res 37:78–84PubMedCrossRefGoogle Scholar
  4. 4.
    Tasci I, Mas MR, Vural SA, Comert B, Alcigir G, Serdar M, Mas N, Isik AT, Ates Y (2006) Rat liver fibrosis regresses better with pegylated interferon alpha2b and ursodeoxycholic acid treatments than spontaneous recovery. Liver Int 26:261–268PubMedCrossRefGoogle Scholar
  5. 5.
    Ryhanen L, Stenback F, Ala-Kokko L, Savolainen ER (1996) The effect of malotilate on type III and type IV collagen, laminin and fibronectin metabolism in dimethylnitrosamine-induced liver fibrosis in the rat. J Hepatol 24:238–245PubMedCrossRefGoogle Scholar
  6. 6.
    Pines M, Knopov V, Genina O, Lavelin I, Nagler A (1997) Halofuginone, a specific inhibitor of collagen type I synthesis, prevents dimethylnitrosamine-induced liver cirrhosis. J Hepatol 27:391–398PubMedCrossRefGoogle Scholar
  7. 7.
    Chen MH, Chen JC, Tsai CC, Wang WC, Chang DC, Tu DG, Hsieh HY (2005) The role of TGF-beta 1 and cytokines in the modulation of liver fibrosis by Sho-saiko-to in rat’s bile duct ligated model. J Ethnopharmacol 97:7–13PubMedCrossRefGoogle Scholar
  8. 8.
    Houglum K, Lee KS, Chojkier M (1997) Proliferation of hepatic stellate cells is inhibited by phosphorylation of CREB on serine 133. J Clin Invest 99:1322–1328PubMedGoogle Scholar
  9. 9.
    Kawada N, Seki S, Inoue M, Kuroki T (1998) Effect of antioxidants, resveratrol, quercetin, and N-acetylcysteine, on the functions of cultured rat hepatic stellate cells and Kupffer cells. Hepatology 27:1265–1274PubMedCrossRefGoogle Scholar
  10. 10.
    Hagiwara SI, Ishii Y, Kitamura S (2000) Aerosolized administration of N-acetylcysteine attenuates lung fibrosis induced by bleomycin in mice. Am J Respir Crit Care Med 162:225–231PubMedGoogle Scholar
  11. 11.
    Pardo A, Ruiz, Arreola JL, Ramirez R, Cisneros-Lira J, Gaxiola M, Barrios R, Kala SV, Lieberman MW, Selman M (2003) Bleomycin-induced pulmonary fibrosis is attenuated in -glutamyl transpeptidase–deficient mice. Am J Respir Crit Care Med 167:925–932PubMedCrossRefGoogle Scholar
  12. 12.
    Serrano-Mollar A, Closa D, Prats N, Blesa S, Martinez-Losa M, Cortijo J, Estrela JM, Morcillo EJ, Bulbena O (2003) In vivo antioxidant treatment protects against bleomycin-induced lung damage in rats. Br J Pharmacol 138:1037–1048PubMedCrossRefGoogle Scholar
  13. 13.
    Vendemiale G, Grattagliano I, Caruso ML, Serviddio G, Valentini AM, Pirrelli M, Altomare E (2001) Increased oxidative stress in dimethylnitrosamine-induced liver fibrosis in the rat: effect of N-acetylcysteine and interferon-alpha. Toxicol Appl Pharmacol 175:130–139PubMedCrossRefGoogle Scholar
  14. 14.
    Tarçın O, Avşar K, Demirtürk L, Gültepe M, Oktar BK, Özdoğan OC, Tarçın Ö, Baloğlu HN, Gürbüz AK (2003) In vivo inefficiency of pentoxifylline and interferon-alpha on hepatic fibrosis in biliary-obstructed rats: assessment by tissue collagen content and prolidase activity. J Gastroenterol Hepatol 18:437–444PubMedCrossRefGoogle Scholar
  15. 15.
    Lopez de Leon A, Rojkind M (1985) A simple micromethod for collagen and total protein determination in formalin-fixed and paraffin-embedded sections. J Histochem Cytochem 33:737–743PubMedGoogle Scholar
  16. 16.
    Knodell RG, Ishak KG, Black WC, Chen TS, Craig R, Kaplowitz N, Kiernan TW, Wollman J (1981) Formulation and application of numerical scoring system for assessing histological activity in asymptomatic chronic active hepatitis. Hepatology 1:431–435PubMedCrossRefGoogle Scholar
  17. 17.
    Yagi K (1984) Assay for blood plasma or serum. Methods Enzymol 105:328–331PubMedGoogle Scholar
  18. 18.
    Davies GR, Simmonds NJ, Stevens TR, Grandison A, Blake DR, Rampton DS (1992) Mucosal reactive oxygen metabolite production in duodenal ulcer disease. Gut 33:1467–1472PubMedCrossRefGoogle Scholar
  19. 19.
    Haklar G, Sayın-Özveri E, Yüksel M, Aktan AÖ, Yalçın AS (2001) Different kinds of reactive oxygen and nitrogen species were detected in colon and breast tumors. Cancer Lett 165:219–224PubMedCrossRefGoogle Scholar
  20. 20.
    Boyne AF, Ellman GL (1972) A methodology for analysis of tissue sulfhydryl components. Anal Biochem 46:639–653PubMedCrossRefGoogle Scholar
  21. 21.
    Geesin JC, Hendricks LJ, Falkenstein PA, Gordon JS, Berg RA (1991) Regulation of collagen synthesis by ascorbic acid: characterization of the role of ascorbate-stimulated lipid peroxidation. Arch Biochem Biophys 290:127–132PubMedCrossRefGoogle Scholar
  22. 22.
    Svegliati Baroni G, D’Ambrosio L, Ferretti G, Casini A, Di Sario A, Salzano R, Ridolfi F, Saccomanno S, Jezequel AM, Benedetti A (1998) Fibrogenic effect of oxidative stress on rat hepatic stellate cells. Hepatology 27:720–726PubMedCrossRefGoogle Scholar
  23. 23.
    Bedossa P, Houglum K, Trautwein C, Holstege A, Chojkier M (1994) Stimulation of collagen alpha 1(I) gene expression is associated with lipid peroxidation in hepatocellular injury: a link to tissue fibrosis? Hepatology 19:1262–1271PubMedGoogle Scholar
  24. 24.
    Lee KS, Buck M, Houglum K, Chojkier M (1995) Activation of hepatic stellate cells by TGF alpha and collagen type I is mediated by oxidative stress through c-myb expression. J Clin Invest 96:2461–2468PubMedCrossRefGoogle Scholar
  25. 25.
    Howard RJMW, Blake DR, Pall H, Williams A, Green ID (1987) Allopurinol/N-acetylcysteine for carbon monoxide poisoning. Lancet 2:628–629PubMedCrossRefGoogle Scholar
  26. 26.
    Ceconi C, Curello S, Cargnoni A, Ferrari R, Albertini A, Visioli O (1988) The role of glutathione status in the protection against ischemic and reperfusion damage: effects of N-acetylcysteine. J Mol Cell Cardiol 20:5–13PubMedCrossRefGoogle Scholar
  27. 27.
    Kalebic T, Kinter A, Poli G, Anderson ME, Meister A, Fauci AS (1991) Suppression of human immunodeficiency virus expression in chronically infected monocytic cells by glutathione, glutathione ester, and N-acetylcysteine. Proc Natl Acad Sci USA 88:986–990PubMedCrossRefGoogle Scholar
  28. 28.
    Bernard GR (1990) Potential of N-acetylcysteine as treatment for the adult respiratory distress syndrome. Eur Respir J 3(supp1):496–498Google Scholar
  29. 29.
    Kawada N, Seki S, Inoue M, Kuroki T (1998) Effect of antioxidants, resveratrol, quercetin, and N-acetylcysteine, on the functions of cultured rat hepatic stellate cells and Kupffer cells. Hepatology 27:1265–1274PubMedCrossRefGoogle Scholar
  30. 30.
    Bataller R, Schwabe RF, Choi YH, Yang L, Paik YH, Lindquist J, Qian T, Schoonhoven R, Hagedorn CH, Lemasters JJ, Brenner DA (2003) NADPH oxidase signal transduces angiotensin II in hepatic stellate cells and is critical in hepatic fibrosis. J Clin Invest 112:1383–1394PubMedCrossRefGoogle Scholar
  31. 31.
    Dogru-Abbasoglu S, Balkan J, Kanbagli O, Cevikbas U, Aykac-Toker G, Uysal M (2002) Aminoguanidine, an inducible nitric oxide synthase inhibitor, plus N-acetylcysteine treatment reduce the lipopolysaccharide-augmented hepatotoxicity in rats with cirrhosis. Hum Exp Toxicol 21:359–364PubMedCrossRefGoogle Scholar
  32. 32.
    Liu JJ, Wang JY, Zhang C, Nilsson A, Duan RD (2002) Hepatic cirrhosis increases sensitivity of kidney to endotoxin in rats. Med Sci Monit 8:BR56–60PubMedGoogle Scholar
  33. 33.
    Tsukamoto H, Horne W, Kamimura S, Niemela O, Parkkila S, Yla-Herttuala S, Brittenham GM (1995) Experimental liver cirrhosis induced by alcohol and iron. J Clin Invest 96:620–630PubMedGoogle Scholar
  34. 34.
    Tsukamoto H (1993) Oxidative stress, antioxidants, and alcoholic liver fibrogenesis. Alcohol 10:465–467PubMedCrossRefGoogle Scholar
  35. 35.
    Fort J, Pilette C, Veal N, Oberti F, Gallois Y, Douay O, Rosenbaum J, Cales P (1998) Effects of long-term administration of interferon alpha in two models of liver fibrosis in rats. J Hepatol 29:263–270PubMedCrossRefGoogle Scholar
  36. 36.
    Shiesh SC, Chen CY, Lin XZ, Liu ZA, Tsao HC (2000) Melatonin prevents pigment gallstone formation induced by bile duct ligation in guinea pigs. Hepatology 32:455–460PubMedCrossRefGoogle Scholar
  37. 37.
    Pastor A, Collado PS, Almar M, Gonzalez-Gallego J (1997) Antioxidant enzyme status in biliary obstructed rats: effects of N-acetylcysteine. J Hepatol 27:363–370PubMedCrossRefGoogle Scholar
  38. 38.
    Cabre M, Camps J, Paternain JL, Ferre N, Joven J (2000) Time-course of changes in hepatic lipid peroxidation and glutathione metabolism in rats with carbon tetrachloride-induced cirrhosis. Clin Exp Pharmacol Physiol 27:694–699PubMedCrossRefGoogle Scholar
  39. 39.
    Lopez PM, Finana IT, De Agueda MC, Sanchez EC, Munoz MC, Alvarez JP, De La Torre Lozano EJ (2000) Protective effect of melatonin against oxidative stress induced by ligature of extra-hepatic biliary duct in rats: comparison with the effect of S-adenosyl-L-methionine. J Pineal Res 28:143–149PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Gulgun Tahan
    • 1
  • Orhan Tarcin
    • 2
  • Veysel Tahan
    • 3
  • Fatih Eren
    • 4
  • Nursal Gedik
    • 5
  • Elife Sahan
    • 6
  • Nilgun Biberoglu
    • 7
  • Savas Guzel
    • 7
  • Aysun Bozbas
    • 3
  • Nurdan Tozun
    • 3
  • Osman Yucel
    • 8
  1. 1.Marmara University Institute of Gastroenterology and Taksim State HospitalDepartment of General SurgeryIstanbulTurkey
  2. 2.Gumussuyu Military HospitalDepartment of GastroenterologyIstanbulTurkey
  3. 3.Marmara UniversityDepartment of GastroenterologyIstanbulTurkey
  4. 4.Marmara UniversityGastroenterology InstituteIstanbulTurkey
  5. 5.Kasimpasa Military HospitalDepartment of BiochemistryIstanbulTurkey
  6. 6.Taksim State HospitalDepartment of PathologyIstanbulTurkey
  7. 7.Taksim State HospitalDepartment of BiochemistryIstanbulTurkey
  8. 8.Taksim State HospitalDepartment of General SurgeryIstanbulTurkey

Personalised recommendations