Advances in Clinical and Experimental Medicine

Title abbreviation: Adv Clin Exp Med
JCR Impact Factor (IF) – 2.1
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ISSN 1899–5276 (print)
ISSN 2451-2680 (online)
Periodicity – monthly

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Advances in Clinical and Experimental Medicine

2007, vol. 16, nr 5, September-October, p. 609–618

Publication type: original article

Language: English

The Role of Calcium Channel−Blocking Drugs in Preserving Rat Liver for Transplantation

Wpływ leków blokujących kanały wapniowe na wątrobę przechowywaną do transplantacji

Małgorzata Trocha1,, Adam Szeląg1,

1 Department of Pharmacology, Silesian Piasts University of Medicine in Wrocław, Poland

Abstract

Background. Liver transplantation has become a widely accepted therapy for patients with end−stage liver diseases. The efficacy of this method depends on many different factors. Calcium plays a crucial role in physiological processes in liver cells; however, in high concentrations its ions can be pathogenic and lead to cell death. Because calcium ions enter the cell mostly through calcium channels, it has been suggested that calcium channel inhibitors (CCIs) could protect hepatocytes from the action of toxic substances.
Objectives. Using an extracorporeal liver perfusion model, the protective actions of nitrendipine and nifedipine during ischemia and reperfusion on the structure and function of rat liver preserved for transplantation were evaluated.
Material and Methods. The study was carried out on livers isolated from adult male Wistar rats. The rat livers were preserved for 24 hours in HTK solution (4ºC) with or without nifedipine or nitrendipine. After preservation, the livers were flushed with Ringer’s solution and perfused for two hours with a perfusion fluid. Glucose concentration and ALT, AST, and LDH activities were measured during perfusion. After perfusion all the produced bile was collected and the livers were histologically examined.
Results. Astatistically significant influence of nitrendipine on the activities of the liver enzymes was found. Livers which were both perfused and rinsed with addition of this drug showed the lowest levels of enzyme activity and histological damage among all the examined groups. No statistically significant differences between the groups in glucose concentration and total amount of bile were observed.
Conclusion. Nitrendipine demonstrated a protective influence on the function and structure of rat liver preserved for transplantation and maintained its action throughout perfusion, especially when added not only to the perfusion fluid, but also to the Ringer’s solution used for rinsing.

Streszczenie

Wprowadzenie. Obecnie przeszczepianie wątroby jest popularną i akceptowaną metodą leczenia pacjentów w ostatnim stadium niewydolności wątroby. Skuteczność tej metody zależy od wielu czynników. Jednym z nich jest prawidłowe stężenie jonów wapnia w tym narządzie. Jeśli nasilenie uszkodzeń w wątrobie podczas jej przechowywania do przeszczepienia zależy od wzrostu stężenia wapnia w komórce, to leki, które hamują jego napływ do komórki, powinny działać cytoprotekcyjnie.
Cel pracy. Ocena cytoprotekcyjnego działania nitrendypiny i nifedypiny na strukturę i czynność wątroby szczurzej przechowywanej do przeszczepienia.
Materiał i metody. Czynność wątroby badano podczas dwugodzinnej perfuzji pozaustrojowej. W grupie zerowej wątroby perfundowano bezpośrednio po izolacji narządu z organizmu, w pozostałych grupach – po 24−godzinnym okresie przechowywania w temperaturze 4ºC w roztworze HTK. W grupach badanych do płynu HTK dodano nitrendypinę lub nifedypinę w stężeniu 20 μmol/l. Dodatkowo, w ostatniej grupie badanej, nitrendypinę w tym samym stężeniu dodano także do roztworu Ringera służącego do przepłukania i ogrzania wątroby. Podczas perfuzji oznaczano aktywność enzymów wątrobowych (ALT, AST i LDH) oraz stężenie glukozy. Pod koniec perfuzji oznaczano całkowitą ilość wydzielonej żółci. W mikroskopie świetlnym oceniano zmiany strukturalne. Liczbę uszkodzonych komórek niemiąższowych oceniano, licząc jądra tych komórek wybarwione błękitem trypanu.
Wyniki. W badaniu wykazano statystycznie istotny wpływ nitrendypiny na aktywność enzymów wątrobowych. W grupie, w której dodano nitrendypinę zarówno do płynu HTK, jak i roztworu Ringera aktywność enzymów była najmniejsza, a zmiany histopatalogiczne najsłabiej wyrażone. Nie wykazano żadnych istotnych różnic w stężeniu glukozy i całkowitej ilości żółci między grupami.
Wnioski. Nitrendypina wykazuje ochronne działanie na czynność i strukturę wątroby szczurzej przechowywanej do przeszczepienia. Działanie to utrzymuje się przez cały okres perfuzji, szczególnie wtedy, gdy została podana zarówno do płynu HTK, jak i roztworu Ringera.

Key words

calcium channel blockers, liver, preservation−reperfusion injury, extracorporeal perfusion

Słowa kluczowe

leki blokujące kanały wapniowe, wątroba, uszkodzenia niedokrwienno−reperfuzyjne, perfuzja pozaustrojowa

References (37)

  1. Gilbert JR, Pascual M, Schoenfeld DA, Rubin RH, Delmonico FL, Cosimi AB: Evolving trends in liver transplantation. Transplantation 1999, 67, 246–253.
  2. Cobreros A, Sainz L, Lasheras B, Cenarruzabeitia E: Hepatotoxicity of ethanol: protective effect of calcium channel blockers in isolated hepatocytes. Liver 1997, 17, 76–82.
  3. De Broin E, Urata K, Giroux L, Lepage R, Huet PM: Effect of calcium antagonists on rat liver during extended cold preservation−reperfusion. Transplantation 1997, 63, 1547–1554.
  4. Jones TE, Morris RG: Pharmacokinetic interaction between tacrolimus and diltiazem. Clin Pharmacokinet 2002, 41, 381–388.
  5. Mies S, Massarollo PC, Figueira ER, Leitao RM, Raia S: Lower incidence of liver graft rejection in patients on diltiazem plus cyclosporine therapy. Transplant Proc 1998, 30, 1437–1438.
  6. Cheng S, Ragsdale JR, Sasaki AW, Lee RG, Deveney CW, Pinson CW: Verapamil improves rat hepatic preservation with UW solution. J Surg Res 1991, 50, 560–564.
  7. Lemasters JJ, DiGiuseppi J, Nieminen AL, Herman B: Blebbing, free Ca2+ and mitochondrial membrane potential preceding cell death in hepatocytes. Nature 1987, 325, 78–81.
  8. Szeląg A, Magdalan J, Rutkowska M, Dziewiszek W, Trocha M, Rzepka M, Pieśniewska M, Fereniec L: Influence of nifedipine, nitrendipine and verapamil at low concentration on antipyrine metabolism examined by extracorporeal rat liver perfusion. Pol J Pharm 2003, 55, 203–208.
  9. Takeda Y, Arii S, Kaido T, Niwano M, Moriga T, Mori A, Hanaki K, Gorrin−Rivas MJ, Ishii T, Sato M, Imamura M: Morphologic alteration of hepatocytes and sinusoidal endothelial cells in rat fatty liver during cold preservation and the protective effect of hepatocyte growth factor. Transplantation 1999, 67, 820–828.
  10. Takei Y, Marzi I, Kauffman FC, Currin RT, Lemasters JJ, Thurman RG: Increase in survival time of liver transplants by protease inhibitors and a calcium channel blocker, nisoldipine. Transplantation 1990, 50, 14–20.
  11. Arai M, Peng XX, Currin RT, Thurman RG, Lemasters JJ: Protection of sinusoidal endothelial cells against storage/reperfusion injury by prostaglandin E2 derived from Kupffer cells. Transplantation 1999, 68, 440–445.
  12. Gao W, Bentley RC, Madden JF, Clavien PA: Apoptosis of sinusoidal endothelial cells is a critical mechanism of preservation injury in rat liver transplantation. Hepatology 1998, 27, 1652–1660.
  13. Schemmer P, Connor HD, Arteel GE, Raleigh JA, Bunzendahl H, Mason RP, Thurman RG: Reperfusion injury in livers due to gentle in situ organ manipulation during harvest involves hypoxia and free radicals. J Pharmacol Exp Ther 1999, 290, 235–240.
  14. Fujita Y, Kimura K, Takaori M: Influence of nicardipine on post−hypoxic injury in the isolated perfused rat liver. Resuscitation 1991, 22, 253–260.
  15. Umeshita K, Monden M, Ukei T, Gotoh M, Nakano Y, Endoh W, Okamura J, Mori T: Different cytoprotective effects of calcium blockers in hypothermic liver preservation. Transplant Proc 1989, 21, 1290–1291.
  16. Iimuro Y, Ikejima K, Rose ML, Bradford BU, Thurman RG: Nimodipine a dihydropyridine−type calcium channel blocker prevents alcoholic hepatitis caused by chronic intragastric ethanol exposure in the rat. Hepatology 1996, 24, 391–397.
  17. Karwinski W, Garcia R, Helton WS: Protective effects of the calcium channel blocker verapamil on hepatic function following warm ischemia. J Surg Res 1999, 64, 150–155.
  18. Liang DC, Thurman RG: Protective effects of the calcium antagonists diltiazem and TA3090 against hepatic injury due to hypoxia. Biochem Pharmacol 1992, 44, 2207–2211.
  19. Gasbarrini A, Borle AB, Van Thiel DH: Ca2+ antagonists do not protect isolated perfused rat hepatocytes from anoxic injury. Biochim Biophys Acta 1993, 1177, 1–7.
  20. Matsuda S: Protective effects of calcium antagonist (nitrendipine) on calcium ionophore A 23187−induced liver cell injury. Bull Tokyo Med Dent Univ 1991, 38, 35–44.
  21. Thurman RG, Apel ED, Lemasters JJ: Protective effect of nitrendipine against hypoxic injury in perfused livers from ethanol−treated rats. J Cardiovasc Pharmacol 1988, 12, Suppl. 4, S113–S116.
  22. Habior A: Liver diseases (Polish). In: Interna. Eds.: Januszkiewicz W, Kokot F, PZWL, Warsaw 2002, 558–591.
  23. Piratvisuth T, Dunne JB, Williams R, Tredger JM: Amlodipine improves hepatic hemodynamic and metabolic function in the isolated perfused rat liver after sequential cold and warm ischemia. Transplantation 1995, 60, 23–28.
  24. Bowers BA, Branum GD, Rotolo FS, Watters CR, Meyers WC: Bile flow – an index of ischaemic injury. J Surg Res 1987, 42, 565–569.
  25. Churchill TA, Green CJ, Fuller BJ: The importance of calcium−related effects on energetics at hypothermia: effects of membrane−channel antagonists on energy metabolism of rat liver. Cryobiology 1995, 32, 477–486.
  26. Bayomi MA, Abanumay KA, al−Angary AA: Effect of inclusion complexation with cyclodextrins on photostability of nifedipine in solid state. Int J Pharm 2002, 243, 107–117.
  27. Haddad P, Cabrillac JC, Piche D, Musallam L, Huet PM: Changes in intracellular calcium induced by acute hypothermia in parenchymal, endothelial, and Kupffer cells of the rat liver. Cryobiology 1999, 39, 69–79.
  28. Lidofsky SD, Xie MH, Sostman A, Scharschmidt BF, Fitz JG: Vasopressin increases cytosolic sodium concentration in hepatocytes and activates calcium influx through cation−selective channels. J Biol Chem 1993, 268, 14632–14636.
  29. Konrad T, Bloechle C, Haller G, Broelsch CE, Usadel KH, Kusterer: Verapamil and flunarizine protect the isolated perfused rat liver against warm ischemia and reperfusion injury. Res Exp Med 1995, 195, 61–68.
  30. Wu J, Danielsson A, Lindstrom P, Karlsson K, Sehlin J: Protective effects of calcium channel blockers on acute bromobenzene toxicity to isolated rat hepatocytes. Inhibition of phenylephrine−induced calcium oscillations. Scand J Gastroenterol 1995, 30, 590–600.
  31. Hijioka T, Rosenberg RL, Lemasters JJ, Thurman RG: Kupffer cells contain voltage−dependent calcium channels. Mol Pharmacol 1992, 41, 435–440.
  32. Kurita K, Tanabe G, Aikou T, Shimazu H: Inhibition of the increase of intrahepatic Ca2+ by diltiazem in rats with liver ischemia. J Hepatol 1994, 21, 567–571.
  33. Michael AD, Whiting RL: Cellular action of nicardipine. Am J Cardiol 1989, 64, 3H–7H.
  34. Stein HJ, Oosthuizen MM, Hinder RA: Effect of verapamil on hepatic ischaemic−reperfusion injury in normal and glutathione−depleted rats. Gastroenterology 1989, 96, A662–A663.
  35. Mak IT, Weglicki WB: Comparative antioxidant activities of propranolol nifedipine verapamil and diltiazem against sarcolemmal membrane lipid peroxidation. Circ Res 1990, 66, 1449–1452.
  36. Crenesse D, Tornieri K, Laurens M, Heurteaux C, Cursio R, Gugenheim J, Schmid−Alliana A: Diltiazem reduces apoptosis in rat hepatocytes subjected to warm hypoxia−reoxygenation. Pharmacology 2002, 65, 87–95.
  37. Abdennebi HB, Steghens JP, Margonari J, Ramella−Virieux S, Barbieux A, Boillot O: Evaluation of parenchymal and nonparenchymal cell injury after different conditions of storage and reperfusion. Transpl Int 1998, 11, 365–372.