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Involvement of Hepatic Stimulator Substance in Experimentally Induced Fibrosis and Cirrhosis in the Rat

  • Georgios Gribilas
  • Apostolos Zarros
  • Athina Zira
  • Costas Giaginis
  • Gerasimos Tsourouflis
  • Charis Liapi
  • Chara Spiliopoulou
  • Stamatios E. Theocharis
Original Article

Abstract

Liver fibrosis results from sustained wound healing response to chronic liver injury. Liver cirrhosis, the end stage of the fibrotic process, is characterized by disruption of the entire liver architecture and reduced hepatocyte regenerative ability. Hepatic stimulator substance (HSS) is a liver-specific growth factor triggering hepatocyte proliferation in vitro and in vivo. Previous studies have indicated the involvement of HSS in animal models of acute liver injury. The aim of the present study was to investigate the involvement of HSS in the process of fibrosis and cirrhosis induction. Liver fibrosis and cirrhosis were induced in rats by thioacetamide (TAA) administration (300 mg/l) in the drinking water for 3 months, and animals were killed at 0, 1, 2, and 3 months of treatment. TAA administration resulted in progressively increasing liver fibrosis, leading to the onset of cirrhosis at the end of the experimental time. HSS was continuously produced during the course of fibrosis and cirrhosis induction, peaking at the 2nd month of TAA treatment, coinciding with markers of hepatic proliferative capacity, as thymidine kinase activity and DNA biosynthesis. Significantly reduced HSS activity was noted in cirrhotic liver (3rd month). In this case, the exogenous HSS administration during the 3rd month of TAA treatment suppressed the onset of liver cirrhosis, stimulating the hepatic regenerative capacity. Our data indicate the active participation of HSS in the process of fibrosis and cirrhosis induction post-TAA treatment in rats, suggesting also the beneficial effect of HSS treatment against cirrhosis induction with future possible clinical implications.

Keywords

HSS Liver Fibrosis Cirrhosis Hepatic regenerative capacity Thioacetamide 

References

  1. 1.
    Michalopoulos GK, De Frances MC. Liver regeneration. Science. 1997;276:60–66. doi: 10.1126/science.276.5309.60.CrossRefPubMedGoogle Scholar
  2. 2.
    Bedossa P, Paradis V. Liver extracellular matrix in health and disease. J Pathol. 2003;200:504–515. doi: 10.1002/path.1397.CrossRefPubMedGoogle Scholar
  3. 3.
    Bataller R, Brenner DA. Liver fibrosis. J Clin Invest. 2005;115:209–218.PubMedGoogle Scholar
  4. 4.
    Tangkijvanich P, Yee HF Jr. Cirrhosis–can we reverse hepatic fibrosis? Eur J Surg Suppl. 2002;587:100–112.PubMedGoogle Scholar
  5. 5.
    La Brecque DR, Pesch LA. Preparation and partial characterization of hepatic regenerative stimulator substance (SS) from rat liver. J Physiol. 1975;248:273–284.Google Scholar
  6. 6.
    La Brecque DR. Hepatic stimulator substance. Discovery, characteristics and mechanism of action. Dig Dis Sci. 1991;36:669–673. doi: 10.1007/BF01297036.CrossRefGoogle Scholar
  7. 7.
    Francavilla A, DiLeo A, Polimeno L, et al. The effect of hepatic stimulatory substance, isolated from regenerating hepatic cytosol, and 50, 000 and 300, 000 subfractions in enhancing survival in experimental acute hepatic failure in rats treated with d-galactosamine. Hepatology. 1986;6:1346–1351. doi: 10.1002/hep.1840060621.CrossRefPubMedGoogle Scholar
  8. 8.
    Yao ZQ, Yang WS, Zhang WB, Chen Y, Zhou YX. Hepatic stimulator substance from human fetal liver for treatment of experimental hepatic failure. Chin Med J (Engl). 1992;105:676–683.Google Scholar
  9. 9.
    Yao ZQ, Yang WS, Zhang WB, Chen YN, Yang FY. Human hepatic regenerative stimulator substance: partial purification and biological characterization of hepatic stimulator substance from human fetal liver cells. Hepatology. 1990;12:1144–1151. doi: 10.1002/hep.1840120512.CrossRefPubMedGoogle Scholar
  10. 10.
    Margeli A, Theocharis S, Spiliopoulou C, Horti M, Koutselinis A. Hepatic stimulator substance administration affects cadmium-induced hepatotoxicity in the rat. Int Hepatol Commun. 1996;5:128–134. doi: 10.1016/0928-4346(96)00290-3.CrossRefGoogle Scholar
  11. 11.
    Theocharis SE, Margeli AP, Spiliopoulou C, Skaltsas S, Kittas C, Koutselinis A. Hepatic stimulator substance administration enhances regenerative capacity of hepatocytes in cadmium-pretreated partially hepatectomized rats. Dig Dis Sci. 1996;41:1475–1480. doi: 10.1007/BF02088575.CrossRefPubMedGoogle Scholar
  12. 12.
    Tzirogiannis KN, Panoutsopoulos GI, Demonakou MD, Hereti RI, Alexandropoulou KN, Mykoniatis MG. Effect of hepatic stimulator substance (HSS) on cadmium-induced acute hepatotoxicity in the rat liver. Dig Dis Sci. 2004;49:1019–1028. doi: 10.1023/B:DDAS.0000034566.43582.53.CrossRefPubMedGoogle Scholar
  13. 13.
    Tzirogiannis KN, Papadimas GK, Kourentzi KT, et al. The role of hepatic stimulator substance (HSS) on liver regeneration arrest induced by cadmium. In Vivo. 2005;19:695–704.PubMedGoogle Scholar
  14. 14.
    Fan QL, Huang CG, Jin Y, et al. Effects of shark hepatic stimulator substance on the function and antioxidant capacity of liver mitochondria in an animal model of acute liver injury. Acta Biochim Biophys Sin. 2005;37:507–514. doi: 10.1111/j.1745-7270.2005.00081.x.CrossRefPubMedGoogle Scholar
  15. 15.
    Margeli AP, Skaltsas SD, Spiliopoulou CA, Mykoniatis MG, Theocharis SE. Hepatic stimulator substance activity in the liver of thioacetamide-intoxicated rats. Liver. 1999;19:519–525. doi: 10.1111/j.1478-3231.1999.tb00085.x.CrossRefPubMedGoogle Scholar
  16. 16.
    Theocharis SE, Margeli AP, Agapitos EV, Mykoniatis MG, Kittas CN, Davaris PS. Effect of hepatic stimulator substance administration on tissue regeneration due to thioacetamide-induced liver injury in rats. Scand J Gastroenterol. 1998;33:656–663. doi: 10.1080/00365529850171954.CrossRefPubMedGoogle Scholar
  17. 17.
    Margeli AP, Manolis E, Skaltsas SN, Tsarpalis KS, Mykoniatis MG, Theocharis SE. Hepatic stimulator substance activity in animal model of fulminant hepatic failure and encephalopathy. Dig Dis Sci. 2002;47:2170–2178. doi: 10.1023/A:1020166706833.CrossRefPubMedGoogle Scholar
  18. 18.
    Margeli AP, Papadimitriou L, Ninos S, Manolis E, Mykoniatis MG, Theocharis SE. Hepatic stimulator substance administration ameliorates liver regeneration in an animal model of fulminant hepatic failure and encephalopathy. Liver Int. 2003;23:171–178. doi: 10.1034/j.1600-0676.2003.00828.x.CrossRefPubMedGoogle Scholar
  19. 19.
    Mei MH, An W, Zhang BH, Shao Q, Gong DZ. Hepatic stimulator substance protects against acute liver failure induced by carbon tetrachloride poisoning in mice. Hepatology. 1993;17:638–644. doi: 10.1002/hep.1840170418.CrossRefPubMedGoogle Scholar
  20. 20.
    Liakos AA, Mykoniatis MG, Kokala ME, Papadimitriou DG, Liatsos GD. Levels of hepatic stimulator substance in liver regenerating process of partially hepatectomized rats pretreated with a single dose of carbon tetrachloride. Dig Dis Sci. 1999;44:1046–1053. doi: 10.1023/A:1026685319823.CrossRefPubMedGoogle Scholar
  21. 21.
    Zhang BH, Gong DZ, Mei MH. Protection of regenerating liver after partial hepatectomy from carbon tetrachloride hepatotoxicity in rats: role of hepatic stimulator substance. J Gastroenterol Hepatol. 1999;14:1010–1017. doi: 10.1046/j.1440-1746.1999.01992.x.CrossRefPubMedGoogle Scholar
  22. 22.
    Liatsos GD, Mykoniatis MG, Margeli A, Liakos AA, Theocharis SE. Effect of acute ethanol exposure on hepatic stimulator substance (HSS) levels during liver regeneration: protective function of HSS. Dig Dis Sci. 2003;48:1929–1938. doi: 10.1023/A:1026157901805.CrossRefPubMedGoogle Scholar
  23. 23.
    Kondili VG, Tzirogiannis KN, Androutsos CD, et al. The hepatoprotective effect of hepatic stimulator substance (HSS) against liver regeneration arrest induced by acute ethanol intoxication. Dig Dis Sci. 2005;50:297–307. doi: 10.1007/s10620-005-1598-9.CrossRefPubMedGoogle Scholar
  24. 24.
    Hwang TL, Yu HC, Chen PC, Chen MF. Liver regeneration following partial hepatectomy and stimulation by hepatic stimulatory substance in cirrhotic and non-cirrhotic rats. Res Exp Med (Berl). 1995;195:201–208. doi: 10.1007/BF02576789.CrossRefGoogle Scholar
  25. 25.
    Zhang M, Song G, Minuk GY. Effects of hepatic stimulator substance, herbal medicine, selenium/vitamin E, and ciprofloxacin on cirrhosis in the rat. Gastroenterology. 1996;110:1150–1155. doi: 10.1053/gast.1996.v110.pm8613004.CrossRefPubMedGoogle Scholar
  26. 26.
    Noda S, Masumi S, Moriyama M, et al. Population of hepatic macrophages and response of perfused liver to platelet-activating factor during production of thioacetamide-induced cirrhosis in rats. Hepatology. 1996;24:412–418. doi: 10.1002/hep.510240220.CrossRefPubMedGoogle Scholar
  27. 27.
    Constantinou MA, Theocharis SE, Mikros E. Application of metabonomics on an experimental model of fibrosis and cirrhosis induced by thioacetamide in rats. Toxicol Appl Pharmacol. 2007;218:11–19. doi: 10.1016/j.taap.2006.10.007.CrossRefPubMedGoogle Scholar
  28. 28.
    Munro HN, Fleck A. Recent developments in the measurement of nucleic acids in biological materials. A supplementary review. Analyst (Lond). 1966;91:78–88. doi: 10.1039/an9669100078.CrossRefGoogle Scholar
  29. 29.
    Richards GM. Modifications of the diphenylamine reaction giving increased sensitivity and simplicity in the estimation of DNA. Anal Biochem. 1974;57:369–376. doi: 10.1016/0003-2697(74)90091-8.CrossRefPubMedGoogle Scholar
  30. 30.
    Kahn D, Svanas GW, Eagon PK, et al. Effect of an antiandrogenic H2 receptor antagonist on hepatic regeneration in rats. J Lab Clin Med. 1988;112:232–239.PubMedGoogle Scholar
  31. 31.
    Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the folin phenol reagent. J Biol Chem. 1951;193:265–275.PubMedGoogle Scholar
  32. 32.
    La Brecque DR, Bachur NR. Hepatic stimulator substance: physicochemical characteristics and specificity. Am J Physiol. 1982;242:G281–G288.Google Scholar
  33. 33.
    Müller A, Machnik F, Zimmermann T, Schubert H. Thioacetamide-induced cirrhosis-like liver lesions in rats–usefulness and reliability of this animal model. Exp Pathol. 1988;34:229–236.PubMedGoogle Scholar
  34. 34.
    Tsukamoto H, Matsuoka M, French SW. Experimental models of hepatic fibrosis: a review. Semin Liver Dis. 1990;10:56–65.CrossRefPubMedGoogle Scholar
  35. 35.
    Kuriyama S, Yokoyama F, Inoue H, et al. Sequential assessment of the intrahepatic expression of epidermal growth factor and transforming growth factor-beta1 in hepatofibrogenesis of a rat cirrhosis model. Int J Mol Med. 2007;19:317–324.PubMedGoogle Scholar
  36. 36.
    Inoue H, Yokoyama F, Kita Y, et al. Relationship between the proliferative capability of hepatocytes and the intrahepatic expression of hepatocyte growth factor and c-Met in the course of cirrhosis development in rats. Int J Mol Med. 2006;17:857–864.PubMedGoogle Scholar
  37. 37.
    MacIntosh E, Gauthier T, Pettigrew N, Minuk G. Liver regeneration and the effect of exogenous putrescine on regenerative activity after partial hepatectomy in cirrhotic rats. Hepatology. 1992;16:1428–1433. doi: 10.1002/hep.1840160620.CrossRefPubMedGoogle Scholar
  38. 38.
    Delhaye M, Louis H, Degraef C, et al. Relationship between hepatocyte proliferative activity and liver functional reserve in human cirrhosis. Hepatology. 1996;23:1003–1011. doi: 10.1002/hep.510230510.CrossRefPubMedGoogle Scholar
  39. 39.
    Delhaye M, Louis H, Degraef C, et al. Hepatocyte proliferative activity in human liver cirrhosis. J Hepatol. 1999;30:461–471. doi: 10.1016/S0168-8278(99)80106-8.CrossRefPubMedGoogle Scholar
  40. 40.
    Fleig WE, Lehmann H, Wagner H, Hoss G, Ditschuneit H. Hepatic regenerative stimulator substance in the rabbit. Relation to liver regeneration after partial hepatectomy. J Hepatol. 1986;3:19–26. doi: 10.1016/S0168-8278(86)80141-6.CrossRefPubMedGoogle Scholar
  41. 41.
    Xia JL, Dai C, Michalopoulos GK, Liu Y. Hepatocyte growth factor attenuates liver fibrosis induced by bile duct ligation. Am J Pathol. 2006;168:1500–1512. doi: 10.2353/ajpath.2006.050747.CrossRefPubMedGoogle Scholar
  42. 42.
    Kusumoto K, Ido A, Moriuchi A, et al. Repeated intravenous injection of recombinant human hepatocyte growth factor ameliorates liver cirrhosis but causes albuminuria in rats. Int J Mol Med. 2006;17:503–509.PubMedGoogle Scholar
  43. 43.
    Lin Y, Xie WF, Chen YX, et al. Treatment of experimental hepatic fibrosis by combinational delivery of urokinase-type plasminogen activator and hepatocyte growth factor genes. Liver Int. 2005;25:796–807. doi: 10.1111/j.1478-3231.2005.01098.x.CrossRefPubMedGoogle Scholar
  44. 44.
    Ueki T, Kaneda Y, Tsutsui H, et al. Hepatocyte growth factor gene therapy of liver cirrhosis in rats. Nat Med. 1999;5:226–230. doi: 10.1038/5593.CrossRefPubMedGoogle Scholar
  45. 45.
    Matsuda Y, Matsumoto K, Yamada A, et al. Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis. Hepatology. 1997;26:81–89. doi: 10.1002/hep.510260111.CrossRefPubMedGoogle Scholar
  46. 46.
    Matsuda Y, Matsumoto K, Ichida T, Nakamura T. Hepatocyte growth factor suppresses the onset of liver cirrhosis and abrogates lethal hepatic dysfunction in rats. J Biochem. 1995;118:643–649.PubMedGoogle Scholar
  47. 47.
    Lotersztajn S, Julien B, Teixeira-Clerc F, Grenard P, Mallat A. Hepatic fibrosis: molecular mechanisms and drug targets. Annu Rev Pharmacol Toxicol. 2005;45:605–628. doi: 10.1146/annurev.pharmtox.45.120403.095906.CrossRefPubMedGoogle Scholar
  48. 48.
    Jiang B, Sawa M, Yamamoto T, Kasai S. Enhancement of proliferation of intrasplenically transplanted hepatocytes in cirrhotic rats by hepatic stimulatory substance. Transplantation. 1997;63:131–135. doi: 10.1097/00007890-199701150-00024.CrossRefPubMedGoogle Scholar
  49. 49.
    An W, Liu XJ, Lei TG, Dai J, Du GG. Growth induction of hepatic stimulator substance in hepatocytes through its regulation on EGF receptors. Cell Res. 1999;9:37–49. doi: 10.1038/sj.cr.7290004.CrossRefPubMedGoogle Scholar
  50. 50.
    Gatzidou E, Kouraklis G, Theocharis S. Insights on augmenter of liver regeneration cloning and function. World J Gastroenterol. 2006;12:4951–4958.PubMedGoogle Scholar
  51. 51.
    Tanigawa K, Sakaida I, Masuhara M, Hagiya M, Okita K. Augmenter of liver regeneration (ALR) may promote liver regeneration by reducing natural killer (NK) cell activity in human liver diseases. J Gastroenterol. 2000;35:112–119. doi: 10.1007/s005350050023.CrossRefPubMedGoogle Scholar
  52. 52.
    Thasler WE, Schlott T, Thelen P, et al. Expression of augmenter of liver regeneration (ALR) in human liver cirrhosis and carcinoma. Histopathology. 2005;47:57–66. doi: 10.1111/j.1365-2559.2005.02172.x.CrossRefPubMedGoogle Scholar
  53. 53.
    Li Q, Liu DW, Zhang LM, Zhu B, He YT, Xiao YH. Effects of augmentation of liver regeneration recombinant plasmid on rat hepatic fibrosis. World J Gastroenterol. 2005;11:2438–2443.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Georgios Gribilas
    • 1
  • Apostolos Zarros
    • 1
    • 2
  • Athina Zira
    • 1
  • Costas Giaginis
    • 1
  • Gerasimos Tsourouflis
    • 1
  • Charis Liapi
    • 2
  • Chara Spiliopoulou
    • 1
  • Stamatios E. Theocharis
    • 1
  1. 1.Department of Forensic Medicine and Toxicology, Medical SchoolNational and Kapodistrian University of AthensAthensGreece
  2. 2.Department of Pharmacology, Medical SchoolNational and Kapodistrian University of AthensAthensGreece

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