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Synergistic effect of erythropoietin but not G-CSF in combination with curcumin on impaired liver regeneration in rats

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The effect of erythropoietin (Epo) and granulocyte colony-stimulating factor (G-CSF) alone or in combination with the hepatoprotective antioxidant curcumin (Cur) was evaluated in a model of delayed liver regeneration.

Materials and methods

Sprague Dawley rats underwent 70% liver resection with simultaneous cecal ligation and puncture and were randomised to five groups: no treatment, G-CSF (100μg/kg), Epo (1,000IU/kg), each alone or in combination with Cur (100mg/kg). Twenty-four hours after surgery, blood and tissue samples were collected. Markers of liver regeneration (liver weight, mitotic index, Ki-67 index), function (bilirubin, bile flow) and hepatocellular damage (liver enzymes, histomorphology) were determined. In addition, cytokine expression and hepatic glutathione concentrations were measured.


Liver regeneration was not improved by G-CSF or Epo monotherapy. Epo more effectively increased liver weight and regeneration markers, but the difference was not significant. Whereas liver regeneration was slightly inhibited in the G-CSF plus Cur group, Epo plus Cur significantly improved liver regeneration. This was accompanied by reduced oxidative stress. Liver function and the expression of pro-inflammatory cytokines were comparable in all treatment groups.


In the present model, Epo, at a relatively low dosage, did not improve liver regeneration. However, the combination of Epo and Cur showed a synergistic effect with highly significant stimulation of liver regeneration.

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cecal ligation and puncture






haematoxylin and eosin (staining)


high-power field




non-parenchymal cells


partial hepatectomy


  1. Joyeux-Faure M (2007) Cellular protection by erythropoietin: new therapeutic implications? J Pharmacol Exp Ther 323:759–762

    Article  PubMed  CAS  Google Scholar 

  2. Brines M, Cerami A (2006) Discovering erythropoietin’s extra-hematopoietic functions: biology and clinical promise. Kidney Int 70:246–250

    Article  PubMed  CAS  Google Scholar 

  3. Lu CZ, Xiao BG (2006) G-CSF and neuroprotection: a therapeutic perspective in cerebral ischaemia. Biochem Soc Trans 34:1327–1333

    Article  PubMed  CAS  Google Scholar 

  4. Lacombe C, Mayeux P (1999) The molecular biology of erythropoietin. Nephrol Dial Transplant 14(Suppl 2):22–28

    Article  PubMed  CAS  Google Scholar 

  5. Farrell F, Lee A (2004) The erythropoetin receptor and its expression in tumor cells and other tissues. Oncologist 9(Suppl 5):18–30

    Article  PubMed  CAS  Google Scholar 

  6. Sakanaka M, Wen TC, Matsuda S, Masuda S, Morishita E, Nagao M, Sasaki R (1998) In vivo evidence that erythropoetin protects neurons from ischemic damage. Proc Natl Acad Sci USA 95:4635–4640

    Article  PubMed  CAS  Google Scholar 

  7. Black D, Lyman S, Heider TR, Behrns KE (2004) Molecular and cellular features of hepatic regeneration. J Surg Res 117:306–315

    Article  PubMed  CAS  Google Scholar 

  8. Cuzzocrea S, Mazzon E, di Paola R, Genovese T, Patel NS, Britti D, de Majo M, Caputi AP, Thiemermann C (2005) Erythropoietin reduces the degree of arthritis caused by type II collagen in the mouse. Arthritis Rheum 52:940–950

    Article  PubMed  CAS  Google Scholar 

  9. Liu X, Xie W, Liu P, Duan M, Jia Z, Li W, Xu J (2006) Mechanism of the cardioprotection of rhEPO pretreatment on suppressing the inflammatory response in ischemia–reperfusion. Life Sci 78:2255–2264

    Article  PubMed  CAS  Google Scholar 

  10. Ates E, Yalcin AU, Yilmaz S, Koken T, Tokyol C (2004) Protective effect of erythropoietin on renal ischemia and reperfusion injury. ANZ J Surg 75:1100–1105

    Article  Google Scholar 

  11. Yilmaz S, Ates E, Tokyol C, Pehlivan T, Erkasap S, Koken T (2004) The protective effect of erythropoietin on ischemia–reperfusion injury of liver. HPB 3:169–173

    Google Scholar 

  12. Calapai G, Marciano MC, Corica F, Allegra A, Parisi A, Frisina N, Caputi AP, Buemi M (2000) Erythropoietin protects against brain ischemic injury by inhibition of nitric oxide formation. Eur J Pharmacol 401:349–356

    Article  PubMed  CAS  Google Scholar 

  13. Baker JE, Kozik D, Hsu AK, Fu X, Tweddell JS, Gross GJ (2007) Darbepoetin alfa protects the rat heart against infarction: dose–response, phase of action, and mechanisms. J Cardiovasc Pharmacol 49:337–345

    Article  PubMed  CAS  Google Scholar 

  14. Ates E, Yilmaz S, Ihtiyar E, Yasar B, Karahuseyinoglu E (2006) Preconditioning-like amelioration of erythropoietin against laparoscopy-induced oxidative injury. Surg Endosc 20:815–819

    Article  PubMed  CAS  Google Scholar 

  15. Le Minh K, Klemm K, Abshagen K, Eipel C, Menger MD, Vollmar B (2007) Attenuation of inflammation and apoptosis by pre- and posttreatment of darbepoetin-alpha in acute liver failure of mice. Am J Pathol 170:1954–1963

    Article  PubMed  CAS  Google Scholar 

  16. Matchett GA, Calinisan JB, Matchett GC, Martin RD, Zhang JH (2007) The effect of granulocyte-colony stimulating factor in global cerebral ischemia in rats. Brain Res 1136:200–207

    Article  PubMed  CAS  Google Scholar 

  17. Bauhofer A, Stinner B, Kohlert F, Reckzeh B, Lorenz W, Celik I (2002) Granulocyte colony-stimulating factor but not peritoneal lavage increases survival rate after experimental abdominal contamination and infection. Br J Surg 89:1457–1464

    Article  PubMed  CAS  Google Scholar 

  18. Bauhofer A, Celik I, Plaul U, Wulf H, Torossian A (2004) Effects of G-CSF and antibiotic prophylaxis in a 2 × 2 factorial design on outcome in septic rats. Inflamm Res 53(Suppl 2):S 126–129

    CAS  Google Scholar 

  19. Görgen I, Hartung T, Leist M, Niehörster M, Tiegs G, Uhlig S, Weitzel F, Wendel A (1992) Granulocyte colony-stimulating factor treatment protects rodents against lipopolysaccharide-induced toxicity via suppression of systemic tumor necrosis factor-alpha. J Immunol 149:918–924

    PubMed  Google Scholar 

  20. Busch CJ, Wanner GA, Menger MD, Vollmar B (2004) Granulocyte colony-stimulating factor (G-CSF) reduces not only gram-negative but also gram-positive infection-associated proinflammatory cytokine release by interaction between Kupffer cells and leukocytes. Inflamm Res 53:205–210

    Article  PubMed  CAS  Google Scholar 

  21. Vollmar B, Messner S, Wanner GA, Hartung T, Menger MD (1997) Immunomodulatory action of G-CSF in a rat model of endotoxin-induced liver injury: an intravital microscopic analysis of Kupffer cell and leukocyte response. J Leukoc Biol 62:710–718

    PubMed  CAS  Google Scholar 

  22. Theocharis SE, Agapitos E, Margeli AP, Goutas N, Kittas C, Davaris P (1997) Effect of two forms of granulocyte-colony stimulating factor on hepatic regeneration after 70% partial hepatectomy in rats. Clin Sci 92:315–320

    PubMed  CAS  Google Scholar 

  23. Inderbitzin D, Beldi G, Sidler D, Studer P, Keogh A, Bisch-Knaden S, Weimann R, Kappeler A, Gloor B, Candinas D (2007) Granulocyte colony-stimulating factor supports liver regeneration in a small-for-size liver remnant mouse model. J Gastrointest Surg 11:280–285

    Article  PubMed  Google Scholar 

  24. Theocharis SE, Papadimitriou LJ, Retsou ZP, Margeli AP, Ninos SS, Papadimitriou JD (2003) Granulocyte-colony stimulating factor administration ameliorates liver regeneration in animal model of fulminant hepatic failure and encephalopathy. Dig Dis Sci 48:1797–1803

    Article  PubMed  CAS  Google Scholar 

  25. Ogiso T, Nagaki M, Takai S, Tsukada Y, Mukai T, Kimura K, Moriwaki H (2007) Granulocyte colony-stimulating factor impairs liver regeneration in mice through the up-regulation of interleukin-1beta. J Hepatol 47:816–25

    Article  PubMed  CAS  Google Scholar 

  26. Seehofer D, Stockmann M, Schirmeier A, Nüssler AK, Cho SR, Rayes N, Koch M, Schiller R, Menger MD, Neuhaus P (2007) Intraabdominal bacterial infections significantly alter regeneration and function of the liver in a rat model of major hepatectomy. Langenbeck’s Arch Surg 392:273–284

    Article  Google Scholar 

  27. Bengmark S (2006) Curcumin, an atoxic antioxidant and natural NFkappaB, cyclooxygenase-2, lipooxygenase, and inducible nitric oxide synthase inhibitor: a shield against acute and chronic diseases. JPEN J Parenter Enteral Nutr 30:45–51

    Article  PubMed  CAS  Google Scholar 

  28. Cho JW, Lee KS, Kim CW (2007) Curcumin attenuates the expression of IL-1beta, IL-6, and TNF-alpha as well as cyclin E in TNF-alpha-treated HaCaT cells; NF-kappaB and MAPKs as potential upstream targets. Int J Mol Med 19:469

    PubMed  CAS  Google Scholar 

  29. Baker MA, Cerniglia GJ, Zaman A (1990) Microtiter plate assay for the measurement of glutathione and glutathione disulfide in large numbers of biological samples. Anal Biochem 190:360

    Article  PubMed  CAS  Google Scholar 

  30. Naughton BA, Birnbach DJ, Liu P, Kolks GA, Tung MZ, Piliero JA, Piliero SJ, Gordon AS (1979) Reticuloendothelial system (RES) hyperfunction and erythropoietin (Ep) production in the regenerating liver. J Surg Oncol 12:227–242

    Article  PubMed  CAS  Google Scholar 

  31. Naughton BA, Gamba-Vitalo C, Naughton GK, Liu P, Gordon AS (1982) Granulopoiesis and colony stimulating factor production in regenerating liver. Exp Hematol 10:451–8

    PubMed  CAS  Google Scholar 

  32. Paul P, Rothmann SA, McMahon JT, Gordon AS (1984) Erythropoietin secretion by isolated rat Kupffer cells. Exp Hematol 12:825–830

    PubMed  CAS  Google Scholar 

  33. Schmeding M, Neumann UP, Boas-Knoop S, Spinelli A, Neuhaus P (2007) Erythropoietin reduces ischemia–reperfusion injury in the rat liver. Eur Surg Res 39:189–97

    Article  PubMed  CAS  Google Scholar 

  34. Schmeding M, Boas-Knoop S, Lippert S, Ruehl M, Somasundaram R, Dagdelen T, Neuhaus P, Neumann UP (2007) Erythropoietin promotes hepatic regeneration after extended liver resection in rats. J Gastroenterol Hepatol DOI 10.1111/j.1440-1746.2007.05265.x

  35. Sasu BJ, Hartley C, Schultz H, McElroy P, Khaja R, Elliott S, Egrie JC, Browne JK, Begley CG, Molineux G (2005) Comparison of epoetin alfa and darbepoetin alfa biological activity under different administration schedules in normal mice. Acta Haematol 113:163–174

    Article  PubMed  CAS  Google Scholar 

  36. Yazihan N, Ataoğlu H, Yener B, Aydin C (2007) Erythropoietin attenuates hydrogen peroxide-induced damage of hepatocytes. Turk J Gastroenterol 18:239–44

    PubMed  Google Scholar 

  37. Alexander WS (2002) Suppressors of cytokine signaling (SOCS) in the immune system. Nat Rev Immunol 2:410–416

    PubMed  CAS  Google Scholar 

  38. Khopde SM, Priyadarsini KI, Venkatesan P, Rao MN (1999) Free radical scavenging ability and antioxidant efficiency of curcumin and its substituted analogue. Biophys Chemist 80:85

    Article  CAS  Google Scholar 

  39. Reddy AC, Lokesh BR (1994) Studies on the inhibitory effects of curcumin and eugenol on the formation of reactive oxygen species and the oxidation of ferrous iron. Mol Cell Biochem 137:1–8

    Article  PubMed  CAS  Google Scholar 

  40. Dickinson DA, Iles KE, Zhang H, Blank V, Forman HJ (2003) Curcumin alters EpRE and AP-1 binding complexes and elevates glutamate-cystein ligase gene expression. FASEB J 17:473

    PubMed  CAS  Google Scholar 

  41. Choi H, Chun YS, Kim SW, Kim MS, Park JW (2006) Curcumin inhibits hypoxia-inducible factor-1 by degrading aryl hydrocarbon receptor nuclear translocator: a mechanism of tumor growth inhibition. Mol Pharmacol 70:1664–1671

    Article  PubMed  CAS  Google Scholar 

  42. Corwin HL, Gettinger A, Fabian TC, May A, Pearl RG, Heard S, An R, Bowers PJ, Burton P, Klausner MA, Corwin MJ (2007) Efficacy and safety of epoetin alfa in critically ill patients. N Engl J Med 357:965–976

    Article  PubMed  CAS  Google Scholar 

  43. Beutel G, Ganser A (2007) Risks and benefits of erythropoiesis-stimulating agents in cancer management. Semin Hematol 44:157–65

    Article  PubMed  CAS  Google Scholar 

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The authors cordially thank Professor Stig Bengmark, University College of London, for his exceptional help during the planning phase of the study and his continuous scientific support during its implementation.

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Correspondence to Daniel Seehofer.

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German Society of Surgery, Surgical Forum 2008, Best of Abstracts.

Daniel Seehofer was supported by grants from the Deutsche Forschungsgemeinschaft (Se1694/2-1).

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Seehofer, D., Neumann, U.P., Schirmeier, A. et al. Synergistic effect of erythropoietin but not G-CSF in combination with curcumin on impaired liver regeneration in rats. Langenbecks Arch Surg 393, 325–332 (2008).

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