Virchows Archiv B

, 64:259 | Cite as

Protective effect of silymarin on rat liver injury induced by ischemia

  • Chuan-Ging Wu
  • R. A. F. M. Chamuleau
  • K. S. Bosch
  • W. M. Frederiks
Original Articles


The effect of silymarin on liver cell damage induced by ischemia was studied in rats fasted for 24 h. In the first series of experiments in vitro ischemia was induced by storing tissue blocks in closed vials at 37°C for 15, 30, 45 and 60 min. Cell injury was detected by the cytophotometrical measurement of glycogen phosphorylase activity in unfixed cryostat sections demonstrated by a modified histochemical procedure. In the second series of experiments in vivo ischemia was provoked by clamping the afferent vessels to the median and left lateral lobes of the liver for 60 min, followed by removal of the clamp and reperfusion. The extent of cell damage was determined by measuring the ALAT and ASAT activities in serum at 1, 3, 6 and 24 h after ischemia and by quantifying the extent of necrosis in the liver after 24 h reperfusion by measuring the unstained areas in cryostat sections incubated for lactate dehydrogenase activity. Silymarin (100mg/kg b.w.) was administered intravenously at 5 min before both the induction of ischemia and the restoration of blood flow (in vivo ischemia). Control received an equal amount of saline. The serum amino-transferase activities after 24 h reperfusion were significantly reduced in the silymarin-treated group (n=10); ALAT 293±193 U/L, ASAT 343±229 U/L compared with the control group (n=7): ALAT 1238±743 U/L, ASAT 948±541 U/L (p<0.03), and the extent of necrosis decreased from 25.6±16.0% (n=7) to 7.8±8.3% (n=10) (p<0.01) after treatment with silymarin. In addition, silymarin appeared to delay and diminish the marked decrease in glycogen phosphorylase activity during 60 min of in vitro ischemia. These results suggest that pretreatment intravenous silymarin can ameliorate hepatic injury associated with ischemia.

Key words

Silymarin Ischemia Liver cell necrosis Glycogen phosphorylase 


  1. Aronson DC, De Haan J, James J, Bosch KS, Ketel AG, Houtkoper JM, Heijmans HSA (1988) Quantitative aspects of the parenchyma-stroma relationship in experimentally induced cholestasis. Liver 8:116–126PubMedGoogle Scholar
  2. Chen JJ, (1990) Flavonoids as superoxide scavengers and antioxidants. Free Radic Biol Med 9:23–28CrossRefGoogle Scholar
  3. Farber JL, Chien KR, Mittnacht S (1981) The pathogenesis of irreversible cell injury in ischemia. Am J Pathol 102:271–281PubMedGoogle Scholar
  4. Feher J (1992) Silymarin as a free radical scavenger in toxic liver lesions (abstract) Free Rad Res Commun [Suppl] D.5Google Scholar
  5. Frederiks WM, James J, Bosch KS, Schroder MJR, Schuyt HC (1982) A model for provoking ischaemic necrosis in rat liver parenchyma and its quantitative analysis. Exp Pathol 22:245–252PubMedGoogle Scholar
  6. Frederiks WM, Myagkaya GL, Fronik GM, Van Veen HA, Vogels IMC, James J (1983) The value of enzyme leakage for the prediction of necrosis in liver ischemia. Histochemistry 78:459–472PubMedCrossRefGoogle Scholar
  7. Frederiks WM, Vogels IMC, Fronik GM (1985) Plasma ornithine carbamyl transferase level as an indication of ischemic injury of rat liver. Cell Biochem Funct 2:217–220CrossRefGoogle Scholar
  8. Frederiks WM, Marx F, Van Noorden CJF (1987) Quantitative histochemical assessment of the heterogeneity of glycogen phosphorylase activity in liver parenchyma of fasted rats using the semipermeable membrane technique and the PAS reaction. Histochem J 19:150–156PubMedCrossRefGoogle Scholar
  9. Frederiks WM, Marx F (1990) The effect of ischemia on glycogen phosphorylase activity in rat liver: a quantitative histochemical study. Anal Cell Pathol 2: 347–353PubMedGoogle Scholar
  10. Granger DN (1988) Role of xanthine oxidase and granulocytes in ischemia-reperfusion injury. Am J Physiol 255:H1269-H1275PubMedGoogle Scholar
  11. Jaeschke H, Farhood A (1991) Neutrophil and Kupffer cell-induced oxidant stress and ischemia-reperfusion injury in rat liver. Am J Physiol 23: G355-G362Google Scholar
  12. Jaeschke H (1991) Reactive oxygen and ischemia/reperfusion injury of the liver. Chem Biol Interact 79:115–136PubMedCrossRefGoogle Scholar
  13. Kunz R, Schoenberg MH, Büchler M, Jost K, Beger HG (1991) Oxygen radicals in liver ischemia and reperfusion. Experimental data. Klin Wochenschr 69:1095–1098PubMedCrossRefGoogle Scholar
  14. Lemasters JJ, Stemkovski GJ, Sungchul JI, Thurman RG (1983) Cell surface changes and enzyme release during hypoxia and reoxygenation in the isolated perfused rat liver. J Cell Biol 97:778–786PubMedCrossRefGoogle Scholar
  15. Lonchampt M, Guardiola B, Sicot N, Bertnrand M, Perdrix L, Duhault JG (1989) Protective effect of a purified flavonoids fraction against reactive oxygen radical. Arzneimittelforschung 39:382–385Google Scholar
  16. Minisci F, Citterio A, Dattilo M (1992) Antioxidant activity of silymarin (abstract). Free Rad Res Commun [Suppl] D.4Google Scholar
  17. Muriel P, Garciapina T, Perez-Alvarez V, Mourelle M (1992) Silymarin protects against paracetamol-induced lipid peroxidation and liver damage. J Appl Toxicol 12:439–442PubMedCrossRefGoogle Scholar
  18. Nishimura T, Yoshida Y, Watanaba F, Koseki M, Nishida T, Tagawa K, Kawashima Y (1986) Blood level of mitochondrial aspartate aminotransferase as an indication of the extent of ischemic necrosis of the rat liver. Hepatology 6:701–707PubMedCrossRefGoogle Scholar
  19. Parks DA, Bulkley GB, Granger DN (1983) Role of oxygen free radicals in shock, ischemia and organ preservation. Surgery 94:428–432PubMedGoogle Scholar
  20. Pascual C, Gonzalez R, Armesto J, Muriel P (1993) Effect of silymarin and silybinin on oxygen radicals. Drug Dev Res 29:73–77CrossRefGoogle Scholar
  21. Van Noorden CJF, Butcher RG (1986) The out of range error in microdensitometry. Histochem J 18:397–398PubMedCrossRefGoogle Scholar
  22. Van Noorden CJF, Frederiks WM (1992) Enzyme histochemistry: a laboratory manual of current method. Royal Microscopical Society, Oxford University PressGoogle Scholar

Copyright information

© Springer-Verlag 1993

Authors and Affiliations

  • Chuan-Ging Wu
    • 1
  • R. A. F. M. Chamuleau
    • 1
  • K. S. Bosch
    • 2
  • W. M. Frederiks
    • 2
  1. 1.Department of Experimental Internal MedicineAcademic Medical CentreAmsterdamThe Netherlands
  2. 2.Laboratory of Cell Biology and HistologyAcademic Medical CentreAmsterdamThe Netherlands

Personalised recommendations