Skip to main content
SpringerLink
Log in

S-nitroso-N-acetylcysteine attenuates liver fibrosis in cirrhotic rats

  • Original Article
  • Published:
Journal of Molecular Medicine Aims and scope Submit manuscript

Abstract

This study was aimed to investigate the molecular mechanisms underlying prevention of hepatic fibrosis by S-nitroso-N-acetylcysteine (SNAC), a nitric oxide donor that inhibits lipid peroxidation. Secondary biliary cirrhosis was induced by 4 weeks of common bile duct ligation (CBDL). Both sham-operated and CBDL animals received SNAC (6.0 μmol/kg/day) starting 2 weeks after surgery. SNAC treatment reduced the increase in blood enzyme activities (alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase), induced by CBDL. Histological changes were attenuated and there was a significant decrease in the area of liver fibrosis and in the activation of stellate cells measured by alpha-smooth muscle actin (α-SMA) immunostaining. The increase in TBARS concentration and hydroperoxide-induced chemiluminescence were also reduced by SNAC treatment. SNAC down-regulated expression of collagen 1α, α-SMA, tumor necrosis factor-α, tumor growth factor-β, metalloproteinase-2, metalloproteinase inhibitor 1, platelet-derived growth factor (PDGF), and PDGF receptor in CBDL rats. These effects were accompanied by inhibited activation of extracellular signal-regulated kinases, Jun amino-terminal kinases, p38 and Akt. Antifibrotic effects were more efficient than those of the free thiol NAC administered at a dose of 60 μmol/kg. In conclusion, results obtained indicate that SNAC, beyond its antioxidant capacity, exerts antifibrotic effects in rats with secondary biliary cirrhosis by down-regulating increased expression of genes and modulating intracellular signaling pathways that contribute to the accumulation of matrix proteins. Thus, SNAC may be an interesting candidate for the treatment of human fibrosis and cirrhosis.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Friedman SL (2008) Mechanisms of hepatic fibrogenesis. Gastroenterology 134:1655–1669

    Article  CAS  PubMed  Google Scholar 

  2. Diaz-Gil JJ, García-Monzón C, Rua C, Martín-Sanz P, Cereceda RM, Miquilena-Colina ME, Machín C, Fernández-Martínez A, García-Cañero R (2008) The antifibrotic effect of liver growth factor is associated with decreased intrahepatic levels of matrix metalloproteinases 2 and 9 and transforming growth factor beta 1 in bile duct-ligated rats. Histol Histopathol 23:583–591

    CAS  PubMed  Google Scholar 

  3. Wynn T (2008) Cellular and molecular mechanisms of fibrosis. J Pathol 214:199–210

    Article  CAS  PubMed  Google Scholar 

  4. Bataller R, Brenner DA (2005) Liver fibrosis. J Clin Invest 115:209–218

    CAS  PubMed  Google Scholar 

  5. Yoshida K, Matsuzaki K, Mori S, Tahashi Y, Yamagata H, Furukawa F, Seki T, Nishizawa M, Fujisawa J, Okazaki K (2005) Transforming growth factor-beta and platelet-derived growth factor signal via c-Jun N-terminal kinase-dependent Smad2/3 phosphorylation in rat hepatic stellate cells after acute liver injury. Am J Pathol 166:1029–1039

    CAS  PubMed  Google Scholar 

  6. Parsons CJ, Takashima M, Rippe RA (2007) Molecular mechanisms of hepatic fibrogenesis. J Gastroenterol Hepatol 22:S79–S84

    Article  CAS  PubMed  Google Scholar 

  7. Geenwel P, Dominguez-Rosales JA, Mavi G, Rivasestilla AM, Rojkind M (2000) Hydrogen peroxide. A link between acetaldehyde-elicited alpha(1) collagen gene up-regulation and oxidative stress in mouse hepatic stellate cells. Hepatology 31:109–116

    Article  Google Scholar 

  8. De Souza GFP, Yokoyama-Yasunaka JKU, Seabra AB, Miguel DC, de Oliveira MG, Uliana SRB (2006) Leishmanicidal activity of primary S-nitrosothiols against Leishmania major and Leishmania amazonensis: implications for the treatment of cutaneous leishmaniasis. Nitric Oxide 15:209–216

    Article  PubMed  Google Scholar 

  9. de Oliveira CP, de Lima VM, Simplicio FI, Soriano FG, de Mello ES, de Souza HP, Alves VA, Laurindo FR, Carrilho FJ, de Oliveira MG (2008) Prevention and reversion of nonalcoholic steatohepatitis in OB/OB mice by S-nitroso-N-acetylcysteine treatment. J Am Coll Nutr 27:299–305

    PubMed  Google Scholar 

  10. de Oliveira CP, Simplicio FI, de Lima VM, Yuahasi K, Lopasso FP, Alves VA, Abdalla DS, Carrilho FJ, Laurindo FR, de Oliveira MG (2006) Oral administration of S-nitroso-N-acetylcysteine prevents the onset of non alcoholic fatty liver disease in rats. World J Gastroenterol 12:1905–1911

    PubMed  Google Scholar 

  11. de Oliveira CP, Stefano JT, de Lima VM, de Sa SV, Simplicio FI, de Mello ES, Corrêa-Giannella ML, Alves VA, Laurindo FR, de Oliveira MG, Giannella-Neto D, Carrilho FJ (2006) Hepatic gene expression profile associated with non-alcoholic steatohepatitis protection by S-nitroso-N-acetylcysteine in ob/ob mice. J Hepatol 45:725–733

    Article  PubMed  Google Scholar 

  12. Kountouras J, Billing BH, Scheuer PJ (1984) Prolonged bile duct obstruction: a new experimental model for cirrhosis in the rat. Br J Exp Pathol 65:305–311

    CAS  PubMed  Google Scholar 

  13. Gonzalez Flecha B, Llesuy S, Boveris A (1991) Hydroperoxide-initiated chemiluminescence: an assay for oxidative stress in biopsies of heart, liver, and muscle. Free Radic Biol Med 10:93–100

    Article  CAS  PubMed  Google Scholar 

  14. Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247:3170–3175

    CAS  PubMed  Google Scholar 

  15. Boveris A, Chance B (1973) The mitochondrial generation of hydrogen peroxide. General properties and effect of hyperbaric oxygen. Biochem J 134:707–716

    CAS  PubMed  Google Scholar 

  16. Moreira A, Fraga C, Alonso P, Collado PS, Zettler C, Marroni C, González-Gallego J (2004) Quercetin prevents B activation in gastric mucosa of portal hypertensive-oxidative stress and NF- rats. Biochem Pharmacol 68:1939–1946

    Article  CAS  PubMed  Google Scholar 

  17. Maeda K, Koda M, Matono T, Sugihara T, Yamamoto S, Ueki M, Murawaki Y, Yamashita N, Nishiyama S (2008) Preventive effects of ME3738 on hepatic fibrosis induced by bile duct ligation in rats. Hepatol Res 38:727–735

    Article  CAS  PubMed  Google Scholar 

  18. Moal F, Veal N, Vuillemin E, Barriere E, Wang J, Oberti F, Gallois Y, Bonnefont-Rousselot D, Rousselet MC, Calès P (2006) Hemodynamic and antifibrotic effects of a selective liver nitric oxide donor V-PYRRO/NO in bile duct ligated rats. World J Gastroenterol 12:6639–6645

    CAS  PubMed  Google Scholar 

  19. Diaz-Gil JJ, García-Monzón C, Rua C, Martín-Sanz C, Cereceda RM, Miquelina-Colina ME, Machín C, Fernández-Martínez A, García-Cañero R (2009) Liver growth factor antifibrotic activity in vivo is associated with decrease in activation of hepatic stellate cells. Histol Histopathol 24:473–479

    CAS  PubMed  Google Scholar 

  20. Liu RM, Liu Y, Forman HJ, Olman M, Tarpey MM (2004) Glutathione regulates transforming growth factor-beta-stimulated collagen production in fibroblasts. Am J Physiol Lung Cell Mol Physiol 286:L121–L128

    Article  CAS  PubMed  Google Scholar 

  21. Nieto N, Friedamn SL, Cederbaum A (2000) Stimulation and proliferation of primary rat hepatic stellate cells by cytochrome P-450 2E-1 derived reactive oxygen species. Hepatology 35:62–73

    Article  Google Scholar 

  22. Gabele E, Froh M, Arteel GE, Uesugi T, Hellerbrand C, Scholmerich J, Brenner DA, Thurman RG, Rippe RA (2009) TNFα is required for cholestasis-induced liver fibrosis in the mouse. Biochem Biophys Res Commun 378:348–353

    Article  CAS  PubMed  Google Scholar 

  23. Failli P, DeFranco R, Caligiuri A, Gentilini A, Romanelli RG, Marra F, Batignani G, Guerra CT, Laffi G, Gentilini P, Pinzani M (2000) Nitrovasodilators inhibit platelet-derived growth factor-induced proliferation and migration of activated hepatic stellate cells. Gastroenterology 119:479–492

    Article  CAS  PubMed  Google Scholar 

  24. Yasuda Y, Shimizu M, Skai H, Iwasa J, Kubota M, Adachi S, Osawa Y, Tsurumi H, Hara Y, Moriwaki H (2009) Epigallocatechin gallate prevents carbon-tetrachloride-induced rat hepatic fibrosis by inhibiting the expression of the PDGFRβ and IGF-1R. Chem Biol Interact 182:159–164

    Article  CAS  PubMed  Google Scholar 

  25. Hemmann S, Graf J, Roderfeld M, Roeb E (2007) Expression of MMPs and TIMPs in liver fibrosis—a systematic review with special emphasis on anti-fibrotic strategies. J Hepatol 46:955–975

    Article  CAS  PubMed  Google Scholar 

  26. Roeb E, Purucker E, BreuerB NH, Heinrich PC, Rose-John S, Matern S (1997) TIMP expression in toxic and cholestatic liver injury in rats. J Hepatol 27:535–544

    Article  CAS  PubMed  Google Scholar 

  27. Sakaida I, Hironaka K, Terai S, Okita K (2003) Gadolinium chloride reverses dimethylnitrosamine (DMN)-induced rat liver fibrosis with increased matrix metalloproteinases (MMPs) of Kupffer cells. Life Sci 72:943–959

    Article  CAS  PubMed  Google Scholar 

  28. Han YP (2006) Matrix metalloproteinases, the pros and cons in liver fibrosis. J Gastroenterol Hepatol 21:S88–S91

    Article  CAS  PubMed  Google Scholar 

  29. Galli A, Svegliati-Baroni G, Ceni E, Milani S, Ridolfi F, Salzano R, Tarocchi M, Grappone C, Pellegrini G, Benedetti A, Surrenti C, Casini A (2005) Oxidative stress stimulates proliferation and invasiveness of hepatic stellate cells via a MMP2-mediated mechanism. Hepatology 41:1074–1084

    Article  CAS  PubMed  Google Scholar 

  30. Qiang H, Lin Y, Zhang X, Zeng X, Shi J, Chen YX, Yang MF, Han ZG, Xie WF (2006) Differential expression genes analyzed by cDNA array in the regulation of rat hepatic fibrogenesis. Liver Int 26:1126–1137

    Article  CAS  PubMed  Google Scholar 

  31. Svegliati-Baroni G, Ridolfi F, Caradonna Z, Alvaro D, Marzionin M, Saccommano S, Candelaresi C, Trozzi L, Macarri G, Benedetti A, Folli F (2003) Regulation of ERK/JNK/p70S6K in two rat models of liver injury and fibrosis. J Hepatol 39:528–537

    Article  CAS  PubMed  Google Scholar 

  32. Rgyu DY, Yang Y, Ha H, Lee GT, Song JS, Uh ST, Lee HB (2005) Role of reactive oxygen species in TGF-β1-induced mitogen-activated protein kinase activation and epithelial mesenchymal transition in renal tubular epithelial cells. J Am Soc Nephrol 16:667–675

    Article  Google Scholar 

  33. Tahan G, Tarcin O, Tahan V, Eren F, Gedik N, Sahan E, Biberoglu N, Guzel S, Bosbaz A, Tozun N, Yucel O (2007) The effects of N-acetylcysteine on bile duct ligation-induced liver fibrosis in rats. Dig Dis Sci 52:3348–3355

    Article  CAS  PubMed  Google Scholar 

  34. Yang YY, Lee KC, Huang YT, Wang YW, Hou MC, Lee FY, Lin HC, Sd L (2008) Effects of N-acetylcysteine administration in hepatic microcirculation of rats with biliary cirrhosis. J Hepatol 49:25–33

    Article  CAS  PubMed  Google Scholar 

  35. Salazar-Montes A, Ruiz-Corro A, López-Reyes A, Castrejón-Gómez E, Armendáriz-Borunda J (2008) Potent antioxidant role of pirfeidone in experimental cirrhosis. Eur J Pharmacol 595:69–77

    Article  CAS  PubMed  Google Scholar 

  36. Baumgardner JN, Shankar K, Hennings L, Albano E, Badger TM, Ronis MJJ (2008) N-acetylcysteine attenuates progression of liver pathology in a rat model of nonalcoholic steatohepatitis. J Nutr 138:1872–1879

    CAS  PubMed  Google Scholar 

  37. Meurer SK, Lahme B, Tihaa L, Weiskirchen R, Gressner AM (2005) N-acetyl-L-cysteine suppresses TGF-β signalling in distinct molecular steps: the biological effcicacy of a multifunctional antifibrotic drug. Biochem Pharmacol 70:1026–1034

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

CIBERehd is funded by the “Instituto de Salud Carlos III”. This work was supported by grants from the Brazilian agencies “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)”, “Fundo de Incentivo à Pesquisa e Eventos (FIPE) do Hospital de Clínicas de Porto Alegre (HCPA)”, and “Laboratório de Hepatologia e Fisiologia Experimental da Universidade Federal do Rio Grande do Sul (HCPA/UFRGS)”.

Conflict of interest statement

The authors have not reported any conflict of interest.

Author information

Authors and Affiliations

  1. Laboratory of Experimental Hepatology and Physiology, Porto Alegre Clinical Hospital, Federal University of Rio Grande do Sul, UFRGS, 90035-903, Ramiro Barcelos, 2350, Porto Alegre, RS, Brazil

    Rafael Vercelino & Norma Possa Marroni

  2. Institute of Biomedicine, University of León and Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Campus of Vegazana s/n, PC 24071, León, Spain

    Irene Crespo, Maria Jose Cuevas, Javier González-Gallego & María Jesús Tuñón

  3. Institute of Chemistry, University of Campinas, UNICAMP, PC 6154, 13083-970, Campinas, SP, Brazil

    Gabriela F. P. de Souza & Marcelo G. de Oliveira

Authors
  1. Rafael Vercelino
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Irene Crespo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gabriela F. P. de Souza
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Maria Jose Cuevas
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marcelo G. de Oliveira
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Norma Possa Marroni
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Javier González-Gallego
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. María Jesús Tuñón
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to María Jesús Tuñón.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vercelino, R., Crespo, I., de Souza, G.F.P. et al. S-nitroso-N-acetylcysteine attenuates liver fibrosis in cirrhotic rats. J Mol Med 88, 401–411 (2010). https://doi.org/10.1007/s00109-009-0577-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00109-009-0577-6

Keywords

Search

Navigation