Synergistic Effects of Jerusalem Artichoke in Combination with Pegylated Interferon Alfa-2a and Ribavirin Against Hepatic Fibrosis in Rats

  • Published : 2016.06.01


Background: Complementary and alternative medicine has been highly appreciated as a supportive regimen for classical treatment strategies. Here we offer a nutrition-based adjuvant therapy for liver fibrosis, a major risk factor for cirrhosis and hepatocellular carcinoma. Aim of the study: To evaluate the possible hepatoprotective effects of Jerusalem artichoke tubers (JAT) in combination with interferon and ribavirin. Materials and Methods: Twelve groups of rats were administered JAT, interferon and ribavirin either separately or in combination from day one of $CCL_4$ administration until the end of the study. Animals were killed after 8 weeks of $CCL_4$-induced hepatotoxicity. Results: Hepatocytes from rats treated with triple combination of interferon, ribavirin, and JAT showed more less normal architecture compared to $CCL_4$-treated rats. We also detected significantly higher hepatic protein expression levels of p53, BAX and transforming growth factor-${\beta}$ (TGF-${\beta}$) in the $CCL_4$-intoxicated group compared to normal controls, as evidenced by immunohistochemical staining and western blotting analyses. Addition of JAT as a supportive regimen improved response to ribavirin and interferon and effectively participated in retaining normal histopathological and biochemical criteria and significantly lowered protein expression of p53, BAX, and TGF-${\beta}$. Conclusions: We suggest that addition of JAT as a supportive r egimen to interferon and ribavirin effectively potentiates their anti-fibrotic effects.


  1. Abdel Salam OM, 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, 2, 19-27.
  2. Abdel-Hamid NM, Nazmy MH, Mahmoud AW, et al (2011). A survey on herbal management of hepatocellular carcinoma. World J Hepatol, 3, 175-83.
  3. Abdel-Hamid NM, Nazmy MH, Wahid A, et al (2015). Jerusalem artichoke attenuates experimental hepatic fibrosis via modulation of apoptotic signaling and fibrogenic activity. Biochemistry Biotechnol Res, 3, 43-50.
  4. Aithal GP, Watkins PB, Andrade RJ, et al (2011). Case definition and phenotype standardization in drug-induced liver injury. Clin Pharmacol Ther, 89, 806-15.
  5. Akyol G, Dursun A, Poyraz A, et al (1999). P53 and proliferating cell nuclear antigen (PCNA) expression in non-tumoral liver diseases. Pathol Int, 49, 214-21.
  6. Althnaian T, Albokhadaim I, El-Bahr SM (2013). Biochemical and histopathological study in rats intoxicated with carbontetrachloride and treated with camel milk. Springerplus, 2, 57.
  7. Arici OF, Cetin N (2011). Protective role of ghrelin against carbon tetrachloride (CCl(4))-induced coagulation disturbances in rats. Regul Pept, 166, 139-42.
  8. Aslan M, Orhan N, Orhan DD, et al (2010). Hypoglycemic activity and antioxidant potential of some medicinal plants traditionally used in Turkey for diabetes. J Ethnopharmacol, 128, 384-9.
  9. Attallah AM, Shiha GE, Ismail H, et al (2009). Expression of p53 protein in liver and sera of patients with liver fibrosis, liver cirrhosis or hepatocellular carcinoma associated with chronic HCV infection. Clin Biochem, 42, 455-61.
  10. Baldo V, Baldovin T, Trivello R, et al (2008). Epidemiology of HCV infection. Curr Pharm Des, 14, 1646-54.
  11. Bataller R, Brenner DA (2005). Liver fibrosis. J Clin Invest, 115, 209-18.
  12. Blight KJ, McKeating JA, Marcotrigiano J, et al (2003). Efficient replication of hepatitis C virus genotype 1a RNAs in cell culture. J Virol, 77, 3181-90.
  13. 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. Hepatol, 26, 643-9.
  14. Brattin WJ, Glende EA, Jr., Recknagel RO (1985). Pathological mechanisms in carbon tetrachloride hepatotoxicity. J Free Radic Biol Med, 1, 27-38.
  15. Castilla A, Prieto J, Fausto N (1991). Transforming growth factors beta 1 and alpha in chronic liver disease. Effects of interferon alfa therapy. N Engl J Med, 324, 933-40.
  16. Chang WC, Jia H, Aw W, et al (2014). Beneficial effects of soluble dietary Jerusalem artichoke (Helianthus tuberosus) in the prevention of the onset of type 2 diabetes and nonalcoholic fatty liver disease in high-fructose diet-fed rats. Br J Nutr, 112, 709-17.
  17. Chang XM, Chang Y, Jia A (2005). Effects of interferon-alpha on expression of hepatic stellate cell and transforming growth factor-beta1 and alpha-smooth muscle actin in rats with hepatic fibrosis. World J Gastroenterol, 11, 2634-6.
  18. CR T (1994). The current role of immunohistochemistry in diagnostic pathology. Adv Pathol Lab Med, 7.
  19. Domitrovic R, Jakovac H, Tomac J, et al (2009). Liver fibrosis in mice induced by carbon tetrachloride and its reversion by luteolin. Toxicol Appl Pharmacol, 241, 311-21.
  20. EA Z (2009). Physiological response to diets fortified with Jerusalem artichoke tubers (Helianthus tuberosus L.) powder by diabetic rats. American-Eurasian J Agric Environ Sci, 5, 682-8.
  21. Fujii H, Nishimura T, Umemura A, et al (2015). Comparison of peg-interferon, ribavirin plus telaprevir vs simeprevir by propensity score matching. World J Hepatol, 7, 2841-8.
  22. Gao B, Bataller R (2011). Alcoholic liver disease: pathogenesis and new therapeutic targets. Gastroenterol, 141, 1572-85.
  23. Gebhardt R (2002). Oxidative stress, plant-derived antioxidants and liver fibrosis. Planta Med, 68, 289-96.
  24. Green DR (2000). Apoptotic pathways: paper wraps stone blunts scissors. Cell, 102, 1-4.
  25. Guo XL, Liang B, Wang XW, et al (2013). Glycyrrhizic acid attenuates CCl(4)-induced hepatocyte apoptosis in rats via a p53-mediated pathway. World J Gastroenterol, 19, 3781-91.
  26. Helm M, Brule H, Giege R, et al (1999). More mistakes by T7 RNA polymerase at the 5 ' ends of in vitro-transcribed RNAs. RNA, 5, 618-21.
  27. Hengartner MO (2000). The biochemistry of apoptosis. Nature, 407, 770-6.
  28. Kucukgergin C, Aydin AF, Ozdemirler-Erata G, et al (2010). Effect of artichoke leaf extract on hepatic and cardiac oxidative stress in rats fed on high cholesterol diet. Biol Trace Elem Res, 135, 264-74.
  29. Lotersztajn S, Julien B, Teixeira-Clerc F, et al (2005). Hepatic fibrosis: molecular mechanisms and drug targets. Annu Rev Pharmacol Toxicol, 45, 605-28.
  30. Mayo MJ (2011). Management of autoimmune hepatitis. Curr Opin Gastroenterol, 27, 224-30.
  31. McCay PB, Lai EK, Poyer JL, et al (1984). Oxygen- and carboncentered free radical formation during carbon tetrachloride metabolism. Observation of lipid radicals in vivo and in vitro. J Biol Chem, 259, 2135-43.
  32. Mehmetcik G, Ozdemirler G, Kocak-Toker N, et al (2008). Role of carnosine in preventing thioacetamide-induced liver injury in the rat. Peptides, 29, 425-9.
  33. Minano C, Garcia-Tsao G (2010). Clinical pharmacology of portal hypertension. Gastroenterol Clin North Am, 39, 681-95.
  34. Muriel P (1996). Alpha-interferon prevents liver collagen deposition and damage induced by prolonged bile duct obstruction in the rat. J Hepatol, 24, 614-21.
  35. Nobili V, Carter-Kent C, Feldstein AE (2011). The role of lifestyle changes in the management of chronic liver disease. BMC Med, 9, 70.
  36. Panasiuk A, Dzieciol J, Panasiuk B, et al (2006). Expression of p53, Bax and Bcl-2 proteins in hepatocytes in non-alcoholic fatty liver disease. World J Gastroenterol, 12, 6198-202.
  37. Papakyriakou P, Tzardi M, Valatas V, et al (2002). Apoptosis and apoptosis related proteins in chronic viral liver disease. Apoptosis, 7, 133-41.
  38. Punzi R, Paradiso A, Fasciano C, et al (2014). Phenols and antioxidant activity in vitro and in vivo of aqueous extracts obtained by ultrasound-assisted extraction from artichoke by-products. Nat Prod Commun, 9, 1315-8.
  39. Ramalho RM, Cortez-Pinto H, Castro RE, et al (2006). Apoptosis and Bcl-2 expression in the livers of patients with steatohepatitis. Eur J Gastroenterol Hepatol, 18, 21-9.
  40. Reitman S, Frankel S (1957). A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol, 28, 56-63.
  41. Riley TR, 3rd, Bhatti AM (2001). Preventive strategies in chronic liver disease: part II. Cirrhosis. Am Fam Physician, 64, 1735-40.
  42. Roberfroid MB, Delzenne NM (1998). Dietary fructans. Annu Rev Nutr, 18, 117-43.
  43. Roberfroid MB, Van Loo JA, Gibson GR (1998). The bifidogenic nature of chicory inulin and its hydrolysis products. J Nutr, 128, 11-9.
  44. Roderfeld M, Geier A, Dietrich CG, et al (2006a). Cytokine blockade inhibits hepatic tissue inhibitor of metalloproteinase-1 expression and up-regulates matrix metalloproteinase-9 in toxic liver injury. Liver Int, 26, 579-86.
  45. Roderfeld M, Weiskirchen R, Wagner S, et al (2006b). Inhibition of hepatic fibrogenesis by matrix metalloproteinase-9 mutants in mice. FASEB J, 20, 444-54.
  46. Sakaida I, Tsuchiya M, Kawaguchi K, et al (2003). Herbal medicine Inchin-ko-to (TJ-135) prevents liver fibrosis and enzyme-altered lesions in rat liver cirrhosis induced by a choline-deficient L-amino acid-defined diet. J Hepatol, 38, 762-9.
  47. Schafer T, Scheuer C, Roemer K, et al (2003). Inhibition of p53 protects liver tissue against endotoxin-induced apoptotic and necrotic cell death. FASEB J, 17, 660-7.
  48. Schuppan D, Afdhal NH (2008). Liver cirrhosis. Lancet, 371, 838-51.
  49. Schuppan D, Jia JD, Brinkhaus B, et al (1999). Herbal products for liver diseases: a therapeutic challenge for the new millennium. Hepatology, 30, 1099-104.
  50. Serni E, Audino V, Del Carlo S, et al (2013). Determination of water-soluble vitamins in multivitamin dietary supplements and in artichokes by micellar electrokinetic chromatography. Nat Prod Res, 27, 2212-5.
  51. Shahnazari P, Sayehmiri K, Minuchehr Z, et al (2014). The increased level of serum p53 in hepatitis b-associated liver cirrhosis. Iran J Med Sci, 39, 446-51.
  52. Solbrig MV, Schlaberg R, Briese T, et al (2002). Neuroprotection and reduced proliferation of microglia in ribavirin-treated bornavirus-infected rats. Antimicrob Agents Chemother, 46, 2287-91.
  53. Tasci I, Mas MR, Vural SA, et al (2007). Pegylated interferonalpha plus taurine in treatment of rat liver fibrosis. World J Gastroenterol, 13, 3237-44.
  54. Thursz M, Yee L, Khakoo S (2011). Understanding the host genetics of chronic hepatitis B and C. Semin Liver Dis, 31, 115-27.
  55. Towbin H, Staehelin T, Gordon J (1979). Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A, 76, 4350-4.
  56. Tsamandas AC, Thomopoulos K, Zolota V, et al (2003). Potential role of bcl-2 and bax mRNA and protein expression in chronic hepatitis type B and C: a clinicopathologic study. Mod Pathol, 16, 1273-88.
  57. Tsushima H, Kawata S, Tamura S, et al (1999). Reduced plasma transforming growth factor-beta1 levels in patients with chronic hepatitis C after interferon-alpha therapy: association with regression of hepatic fibrosis. J Hepatol, 30, 1-7.
  58. Veillon P, Fouchard-Hubert I, Larrey D, et al (2015). Does epoetin beta still have a place in peginterferon alpha-2a plus ribavirin treatment strategies for chronic hepatitis c? J Interferon Cytokine Res.
  59. Vogelstein B, Lane D, Levine AJ (2000). Surfing the p53 network. Nature, 408, 307-10.
  60. Vousden KH, Lu X (2002). Live or let die: the cell’s response to p53. Nat Rev Cancer, 2, 594-604.
  61. Watson D, Rogers JA (1961). A study of six representative methods of plasma bilirubin analysis. J Clin Pathol, 14, 271-8.
  62. Xu XB, Leng XS, Yang X, et al (2004). Obstruction of TGF-beta1 signal transduction can decrease the process of hepatocellular carcinoma in mice induced by CCl4/ethanol. Zhonghua Yi Xue Za Zhi, 84, 1122-5 (in Chinese).
  63. Zawistowski J, Blank G, Murray ED (1986). Polyphenol oxidase activity in Jerusalem artichokes (Helianthus tuberosus L.). Can Institute Food Science Technol J, 19, 210-4.