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Resveratrol inhibits cell growth by inducing cell cycle arrest in activated hepatic stellate cells

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Abstract

Resveratrol (RSV) exerts anti-proliferative and pro-apoptotic actions in different cell lines. Hepatic stellate cells (HSCs) are major fibrogenic cell types that contribute to collagen accumulation during chronic liver disease. In the present study, the inhibitory effects of RSV on cell proliferation, cell cycle, and apoptosis were evaluated in the mouse hepatic stellate cell line GRX. Cells treated with 1 nM–1 μM of RSV demonstrated a decrease in cell growth of about 35% after 5 days. GRX cells, treated with RSV (100 nM or 1 μM), were analyzed by flow cytometry; RSV induced an increase in the number of GRX cells in the S- and sub-G1 phases. The increase in sub-G1 phase cells and the nuclear condensation and fragmentation shown by DAPI staining identified a possible pro-apoptotic effect of RSV on GRX cells. Furthermore, the RSV anti-proliferative effects could be explained by an S-phase accumulation caused by a decrease in the progression through the cell cycle or an inhibition of S or G2 phase transition. It is notable that these RSV actions are mediated at nanomolar levels, compatible with the concentrations of free RSV in biological fluids after ingestion of polyphenol-rich foods, suggesting a possible effect of these foods as an adjuvant treatment in chronical liver diseases.

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References

  1. Pinzani M, Rombouts K (2004) Liver fibrosis: from the bench to clinical targets. Dig Liver Dis 36:231–242

    Article  PubMed  CAS  Google Scholar 

  2. Friedman SL, Roll FJ, Boyles J, Bissell DM (1985) Hepatic lipocytes: the principal collagen-producing cells of normal rat liver. Proc Natl Acad Sci USA 82:8681–8685

    Article  PubMed  CAS  Google Scholar 

  3. Gutierrez-Ruiz MC, Gomez-Quiroz LE (2007) Liver fibrosis: searching for cell model answers. Liver Int 27:434–439

    Article  PubMed  CAS  Google Scholar 

  4. Pinzani M, Gentilini P (1999) Biology of hepatic stellate cells and their possible relevance in the pathogenesis of portal hypertension in cirrhosis. Semin Liver Dis 19:397–410

    Article  PubMed  CAS  Google Scholar 

  5. Borojevic R, Monteiro AN, Vinhas SA, Domont GB, Mourao PA, Emonard H, Jr.Grimaldi G, Grimaud JA (1985) Establishment of a continuous cell line from fibrotic schistosomal granulomas in mice livers. In vitro Cell Dev Biol 21:382–390

    Article  PubMed  CAS  Google Scholar 

  6. Guma FCR, Mello TG, Mermelstein CS, Fortuna VA, Wofchuk ST, Gottfried C, Guaragna RM, Costa ML, Borojevic R (2001) Intermediate filaments modulation in an in vitro model of the hepatic stellate cell activation or conversion into the lipocyte phenotype. Biochem Cell Biol 79:409–417

    Article  PubMed  CAS  Google Scholar 

  7. Margis R, Borojevic R (1989) Retinoid-mediated induction of the fat-storing phenotype in a liver connective tissue cell line (GRX). Biochim Biophys Acta 1011:1–5

    Article  PubMed  CAS  Google Scholar 

  8. Borojevic R, Guaragna RM, Margis R, Dutra HS (1990) In vitro induction of the fat-storing phenotype in a liver connective tissue cell line-GRX. In vitro Cell Dev Biol 26:361–368

    Article  PubMed  CAS  Google Scholar 

  9. Martucci RB, Ziulkoski AL, Fortuna VA, Guaragna RM, Guma FC, Trugo LC, Borojevic R (2004) Beta-carotene storage, conversion to retinoic acid, and induction of the lipocyte phenotype in hepatic stellate cells. J Cell Biochem 92: 414–423

    Article  PubMed  CAS  Google Scholar 

  10. da Silva FM, Guimaraes EL, Grivicich I, Trindade VM, Guaragna RM, Borojevic R, Guma FC (2003) Hepatic stellate cell activation in vitro: cell cycle arrest at G2/M and modification of cell motility. J Cell Biochem 90: 387–396

    Article  PubMed  CAS  Google Scholar 

  11. Chiarini LB, Borojevic R, Monteiro AN (1994) In vitro collagen synthesis by liver connective tissue cells isolated from schistosomal granulomas. Braz J Med Biol Res 27: 1193–1197

    PubMed  CAS  Google Scholar 

  12. Guimaraes EL, Franceschi MF, Grivicich I, Dal-Pizzol F, Moreira JC, Guaragna RM, Borojevic R, Margis R, Guma FC (2006) Relationship between oxidative stress levels and activation state on a hepatic stellate cell line. Liver Int 26:477–485

    Article  PubMed  CAS  Google Scholar 

  13. Donnelly LE, Newton R, Kennedy GE, Fenwick PS, Leung RH, Ito K, Russell RE, Barnes PJ (2004) Anti-inflammatory effects of resveratrol in lung epithelial cells: molecular mechanisms. Am J Physiol Lung Cell Mol Physiol 287:L774–783

    Article  PubMed  CAS  Google Scholar 

  14. Baur JA, Sinclair DA (2006) Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov 5:493–506

    Article  PubMed  CAS  Google Scholar 

  15. Kundu JK, Surh YJ (2004) Molecular basis of chemoprevention by resveratrol: NF-kappaB and AP-1 as potential targets. Mutat Res 555:65–80

    PubMed  CAS  Google Scholar 

  16. Delmas D, Lancon A, Colin D, Jannin B, Latruffe N (2006) Resveratrol as a chemopreventive agent: a promising molecule for fighting cancer. Curr Drug Targets 7:423–442

    Article  PubMed  CAS  Google Scholar 

  17. Zhang W, Fei Z, Zhen HN, Zhang JN, Zhang X (2007) Resveratrol inhibits cell growth and induces apoptosis of rat C6 glioma cells. J Neurooncol 81:231–240

    Article  PubMed  CAS  Google Scholar 

  18. Muir D, Varon S, Manthorpe M (1990) An enzyme-linked immunosorbent assay for bromodeoxyuridine incorporation using fixed microcultures. Anal Biochem 185:377–382

    Article  PubMed  CAS  Google Scholar 

  19. 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 27:1265–1274

    Article  PubMed  CAS  Google Scholar 

  20. Peterson GL (1977) A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem 83:346–356

    Article  PubMed  CAS  Google Scholar 

  21. Hayon T, Dvilansky A, Shpilberg O, Nathan I (2003) Appraisal of the MTT-based assay as a useful tool for predicting drug chemosensitivity in leukemia. Leuk Lymphoma 44:1957–1962

    Article  PubMed  CAS  Google Scholar 

  22. Hansen MB, Nielsen SE, Berg K (1989) Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J Immunol Methods 119:203–210

    Article  PubMed  CAS  Google Scholar 

  23. Chen A, Zhang L, Xu J, Tang J (2002) The antioxidant (−)-epigallocatechin-3-gallate inhibits activated hepatic stellate cell growth and suppresses acetaldehyde-induced gene expression. Biochem J 368:695–704

    Article  PubMed  CAS  Google Scholar 

  24. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279

    Article  PubMed  CAS  Google Scholar 

  25. Bhat KPL, Kosmeder JW II, Pezzuto JM (2001) Biological effects of resveratrol. Antioxid Redox Signal 3:1041–1064

    Google Scholar 

  26. Notas G, Nifli AP, Kampa M, Vercauteren J, Kouroumalis E, Castanas E (2006) Resveratrol exerts its antiproliferative effect on HepG2 hepatocellular carcinoma cells, by inducing cell cycle arrest, and NOS activation. Biochim Biophys Acta 1760:1657–1666

    PubMed  CAS  Google Scholar 

  27. Stervbo U, Vang O, Bonnesen C (2006) Time- and concentration-dependent effects of resveratrol in HL-60 and HepG2 cells. Cell Prolif 39:479–493

    Article  PubMed  CAS  Google Scholar 

  28. Taub R (2004) Liver regeneration: from myth to mechanism. Nat Rev Mol Cell Biol 5:836–847

    Article  PubMed  CAS  Google Scholar 

  29. Balabaud C, Bioulac-Sage P, Desmouliere A (2004) The role of hepatic stellate cells in liver regeneration. J Hepatol 40:1023–1026

    Article  PubMed  CAS  Google Scholar 

  30. Kisseleva T, Brenner DA (2006) Hepatic stellate cells and the reversal of fibrosis. J Gastroenterol Hepatol 21Suppl 3:S84–87

    Article  PubMed  CAS  Google Scholar 

  31. de Aguirres AB, Mello PA, Andrade CM, Breier AC, Margis R, Guaragna RM, Borojevic R, Guma FC, Trindade VM (2007) Variations of ganglioside biosynthetic pathways in the phenotype conversion from myofibroblasts to lipocytes in murine hepatic stellate cell line. Mol Cell Biochem 303:121–130

    Article  PubMed  CAS  Google Scholar 

  32. Andrade CM, Trindade VM, Cardoso CC, Ziulkoski AL, Trugo LC, Guaragna RM, Borojevic R, Guma FC (2003) Changes of sphingolipid species in the phenotype conversion from myofibroblasts to lipocytes in hepatic stellate cells. J Cell Biochem 88:533–544

    Article  PubMed  CAS  Google Scholar 

  33. Mermelstein CS, Guma FC, Mello TG, Fortuna VA, Guaragna RM, Costa ML, Borojevic R (2001) Induction of the lipocyte phenotype in murine hepatic stellate cells: reorganisation of the actin cytoskeleton. Cell Tissue Res 306:75–83

    Article  PubMed  CAS  Google Scholar 

  34. Vicente CP, Fortuna VA, Margis R, Trugo L, Borojevic R (1998) Retinol uptake and metabolism, and cellular retinol binding protein expression in an in vitro model of hepatic stellate cells. Mol Cell Biochem 187:11–21

    Article  PubMed  CAS  Google Scholar 

  35. Vicente CP, Guaragna RM, Borojevic R (1997) Lipid metabolism during in vitro induction of the lipocyte phenotype in hepatic stellate cells. Mol Cell Biochem 168:31–39

    Article  PubMed  CAS  Google Scholar 

  36. dos Santos AQ, Nardin P, Funchal C, de Almeida LM, Jacques-Silva MC, Wofchuk ST, Goncalves CA, Gottfried C (2006) Resveratrol increases glutamate uptake and glutamine synthetase activity in C6 glioma cells. Arch Biochem Biophys 453:161–167

    Article  PubMed  CAS  Google Scholar 

  37. Bertelli A, Bertelli AA, Gozzini A, Giovannini L (1998) Plasma and tissue resveratrol concentrations and pharmacological activity. Drugs Exp Clin Res 24:133–138

    PubMed  CAS  Google Scholar 

  38. Ahmad N, Adhami VM, Afaq F, Feyes DK, Mukhtar H (2001) Resveratrol causes WAF-1/p21-mediated G(1)-phase arrest of cell cycle and induction of apoptosis in human epidermoid carcinoma A431 cells. Clin Cancer Res 7:1466–1473

    PubMed  CAS  Google Scholar 

  39. Park JW, Choi YJ, Jang MA, Lee YS, Jun DY, Suh SI, Baek WK, Suh MH, Jin IN, Kwon TK (2001) Chemopreventive agent resveratrol, a natural product derived from grapes, reversibly inhibits progression through S and G2 phases of the cell cycle in U937 cells. Cancer Lett 163:43–49

    Article  PubMed  CAS  Google Scholar 

  40. Kuo PL, Chiang LC, Lin CC (2002) Resveratrol- induced apoptosis is mediated by p53-dependent pathway in Hep G2 cells. Life Sci 72:23–34

    Article  PubMed  CAS  Google Scholar 

  41. Joe AK, Liu H, Suzui M, Vural ME, Xiao D, Weinstein IB (2002) Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines. Clin Cancer Res 8:893–903

    PubMed  CAS  Google Scholar 

  42. Ragione FD, Cucciolla V, Borriello A, Pietra VD, Racioppi L, Soldati G, Manna C, Galletti P, Zappia V (1998) Resveratrol arrests the cell division cycle at S/G2 phase transition. Biochem Biophys Res Commun 250:53–58

    Article  PubMed  CAS  Google Scholar 

  43. Rigolio R, Miloso M, Nicolini G, Villa D, Scuteri A, Simone M, Tredici G (2005) Resveratrol interference with the cell cycle protects human neuroblastoma SH-SY5Y cell from paclitaxel-induced apoptosis. Neurochem Int 46:205–211

    Article  PubMed  CAS  Google Scholar 

  44. Chen C, Zhang J, Li J, Huang J, Yang C, Huang G, Shi J (2004) Hydrodynamic-based in vivo transfection of retinoic X receptor-alpha gene can enhance vitamin A-induced attenuation of liver fibrosis in mice. Liver Int 24:679–686

    Article  PubMed  CAS  Google Scholar 

  45. Kuwajerwala N, Cifuentes E, Gautam S, Menon M, Barrack ER, Reddy GP (2002) Resveratrol induces prostate cancer cell entry into s phase and inhibits DNA synthesis. Cancer Res 62:2488–2492

    PubMed  CAS  Google Scholar 

  46. Schneider Y, Vincent F, Duranton B, Badolo L, Gosse F, Bergmann C, Seiler N, Raul F (2000) Anti-proliferative effect of resveratrol, a natural component of grapes and wine, on human colonic cancer cells. Cancer Lett 158:85–91

    Article  PubMed  CAS  Google Scholar 

  47. Clement MV, Hirpara JL, Chawdhury SH, Pervaiz S (1998) Chemopreventive agent resveratrol, a natural product derived from grapes, triggers CD95 signaling-dependent apoptosis in human tumor cells. Blood 92:996–1002

    PubMed  CAS  Google Scholar 

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Acknowledgments

This research was supported by CNPq and CAPES grants of the Brazilian Federal Government, FAPERGS grant of the Rio Grande do Sul State Government.

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Authors and Affiliations

  1. Departamento de Bioquímica, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600-anexo, CEP 90035-003, Porto Alegre, RS, Brazil

    Izabel C. Souza, Leo Anderson M. Martins, Barbara P. Coelho, Regina M. Guaragna, Carmem Gottfried & Fátima Costa Rodrigues Guma

  2. ULBRA, Canoas, RS, Brazil

    Izabel C. Souza & Ivana Grivicich

  3. Departamento de Histologia e Embriologia, ICB, PABCAM, UFRJ, Rio de Janeiro, RJ, Brazil

    Radovan Borojevic

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  1. Izabel C. Souza
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  2. Leo Anderson M. Martins
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  5. Regina M. Guaragna
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  8. Fátima Costa Rodrigues Guma
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Correspondence to Fátima Costa Rodrigues Guma.

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Souza, I.C., Martins, L.A.M., Coelho, B.P. et al. Resveratrol inhibits cell growth by inducing cell cycle arrest in activated hepatic stellate cells. Mol Cell Biochem 315, 1–7 (2008). https://doi.org/10.1007/s11010-008-9781-x

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  • DOI: https://doi.org/10.1007/s11010-008-9781-x

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