Changes in Membrane Enzymes in Warm Liver Ischemia

Czech version

Authors: M. Vician; J. Olejník; M. Gergel; I. Brychta; P. Michalka ^*; J. Jakubovský ^*
Authors‘ workplace: Ústav patologickej anatómie LF UKo, Bratislava, Slovenská republika ; Chirurgická klinika SZU a FNsP, nemocnica akad. L. Dérera, Bratislava, Slovenská republika
Published in: Rozhl. Chir., 2010, roč. 89, č. 10, s. 647-653.
Category: Monothematic special - Original

Overview

Background:
The aim of the study is to perform a morphological analysis of certain membrane enzymes and parenchymal alterations during warm ischemia in the pig liver.

Methodology:
Standard hepatectomies were performed in a set of 24 pigs. Intraoperative intravenous (portal vein) hydroxyl radical scavenger Stobadine and Pentoxifylline was administered. Tissue specimens were removed from the margo acutum in interval 10, 60, 80 et 100 minutes. Applied methods of morphological analysis were concentrated on studying the activity of enzymes, which are regarded as cell membrane condition markers. First of them was the alkaline phosphatase, the second studied enzyme was the 5´-mononucleotidase. Detected findings in control group were compared to other two groups. We evaluated each sample using Image Forge Version 1.1 software (PROVER LTD., Bratislava). Applying this software we measured percentage of pre-measured activity levels of both enzymes.

Results:
In normal pig liver, the activity of alkaline phosphatase was regularly distributed in sinusoid walls in the whole lobule. At 10 minutes after the beginning of the warm ischemia, we have found focal losses of enzyme activity, especially in cells of zone 3 of the liver lobuli. One hour after the beginning of warm ischemia were the local losses of enzyme activity even more pronounced. Decreased bile canalicular 5´-mononucleotidase activity reflects cell damage, especially in pericentral areas after 60 min. of ischemia and a further decrease in activity at all surfaces of the membrane. Total amount of enzyme activity is different.

Conclusions:
Morhological findings of enzyme activities showed that zone 2 and 3 of the liver lobule are essencial for the organ survival. The study showed, that intravenous administration of Pentoxifylline and Stobadine probably protects the liver from warm ischemia injury.

Key words:
warm ischemia – pig liver – alkaline phosphatase – 5´-mononucleotidase – Pentoxiphylline – antioxidant

Sources

1. Dixon, E., Vollmer, C. M., Bathe, O. F., et al. Vascular occlusion to decrease blood loss during hepatic resection. Am. J. Surg., 2005; 190: 75–86.

2. Habib, N. A., Černý, J., Danninger, F. Total vascular exclusion of the liver – an advance in hepatic surgery. Rozhl. Chir., 1994; 73: 221–222.

3. Nuzzo, G., Giuliante, F., Vellone, M. Pedicle clamping with ischemic preconditioning in liver resection. Liver Transplant., 2004;10: S53–S57.

4. Serracino-Inglott, F., Habib, N. A., Mathie, R. T. Hepatic ischemia – reperfusion injury. Am. J. Surg., 2001;181: 160–166.

5. Vician, M., Olejník, J., Michalka, P., et al. Warm liver ischemia in experiment and lysosomal markers. Bratisl. Lek. Listy, 2009;110: 587–591.

6. Kiszely, G., Pósalaky, Z. Mikrotechnische und Histochemische Untersuchungsmethoden. Budapest, Akadémiai kiadó, 1964, 724 s.

7. Lojda, Z., Papousek, F. Basics of the enzyme histochemical detection. Brno, UDV- ZSZ, 1970: 68–74.

8. Bancroft, J. D. Theory and practice of histochemical techniques. 5th Ed. Edinburgh, London, Churchill Livingstone, 2002, 800 s.

9. Keselman, H. J., Leventhal, L. Concerning the statistical procedures enumerated by Gentile et al.: Another perspective. Journal of applied behavior analysis, 1974; 7: 643–645.

10. http://www.cs.cmu.edu/afs/cs/project/jair/pub/volume24/ortizboyer05a-html/node14.html, 2006.

11. Horáková, Ľ., Štolc, S. Antioxidant and pharmacodynamic effects of pyridoindol Stobadine. Gen. Pharmac., 1998;30: 627–638.

12. Semmler, J., Gebert, U., Eisenhut, et al. Xantine derivates: comparison between suppression of tumor necrosis factor-alpha production and inhibition of cAMP phosphodiesterase activity. Immunology, 1993; 78: 520–525.

13. Sinha, B., Semmler, J., Eisenhut, T., et al. Enhanced tumor necrosis factor suppression and cyclic adenosine monophosphate accumulation by combination of phosphodiesterase inhibitors and prostanoids. Eur. J. Immunol., 1995; 25: 147–153.

14. Zabel, P., Schade, F. U., Schlaak, M. Inhibition of endogenous TNF formation by pentoxiphylline. Immunobiology, 1993; 187: 447–463.

15. Rudiger, H. A., Clavien, P. A. Tumor necrosis factor-alpha, but not Fas, mediates hepatocellular apoptosis in the murine ischemic liver. Gastroenterology, 2002;122: 202–210.

16. Vajdová, K., Smreková, R., Kukan, M., et al. Endotoxin-induced aggravation of preservation-reperfusion injury of rat liver and its modulation. J. Hepatol., 2000;32: 112–120.

17. Giannini, E. G., Testa, R., Savarino, V. Liver enzyme alteration: a guide for clinicians. CMAJ, Canada‘s Leading Medical Journal, 2005; 172: 367–379.

18. Vokurka, J. Influence of Electric Current on Small Intestine of the Rat. Acta Vet Brno, 2003; 72: 553–558.

19. Vokurka, J. Influence of Electric Current on the Liver Parenchyma of the Rat. Acta Vet. Brno, 2003; 72: 547–551.

20. Vokurka, J. Effect of Gemcitabine on Vascular Endothelium in the Rat. Acta Vet. Brno, 2004;73: 201–204.

21. Koudstaal, J., Hardonk, M. J. A histochemical study about changes in rat liver plasma membrane enzyme activities after galactosamine administration. Histochemistry, 1979; 62: 77–84.

22. Koudstaal, J., Runsink, A. P., van der Sandt, M., Hardonk, M. J. Correlation between serum alkaline phosphatase and localization of alkaline phosphatase in the liver. Acta Histochem., 1975; 14: 129–138.

23. Valko, M., Leibfritz, D., Moncol, J., et al. Free radicals and antioxidants in normal physiological functions and human disease. Int. J. Biochem. Cell Biol., 2007; 39: 44–84.

24. Fabia, R., Travis, D. L., Levy, M. F., et al. Effect of pentoxiphylline on hepatic ischemia and reperfusion injury. Surgery, 1997; 121: 520–525.

25. Pecháň, I., Čornak, V., Rendeková, et al. Mechanisms of the allopurinol on the metabolisms of adenine nucleotides. Bratisl. Lek. Listy,1991; 92: 323–329.

26. Frederiks, W. M., Marx, F., Myagkaya, G. L. A quantitative histochemical study of 5-nucleotidase activity in rat liver after ischemia. Journal of Pathology, 1988; 154: 277–286.

27. Sanada, S., Asanuma, H., Minamino, T., et al. Optimal windows of statin use for immediate infarct limitation 5-nucleotidase as another downstream Molecule of phosphatidylinositol 3-kinase. Circulation, 2004; 110: 2143–2149.

28. Bilčíková, L., Bauer, V., Kolena, J. The effects of methylxantines ethymizol, ephedrine and papaverine on guinea pig and dog trachea. Gen. Physiol. Biophys., 1987; 6: 137–148.