Preferencje help
Widoczny [Schowaj] Abstrakt
Liczba wyników
2011 | 55 | 3 |
Tytuł artykułu

Cytotoxicity studies of lasalocid and silibinin in rat hepatoma cell culture

Warianty tytułu
Języki publikacji
The cvtoprotective effect of silibinin in course of cytotoxicity induced by lasalocid had been measured in rat hepatoma FaO cell line. In the course of the study, MTT test (cellular metabolism), coomassie brillant blue binding test - CBB (total cellular proteins), and LDH release test (membrane integrity) were applied. In addition, changes in the cell morphology after 24 h treatment were observed by light microscopy. The effective concentrations, EC₅₀ were quantified for each compound alone, whereas the nature of their co-action was assessed by isobologram plotting. Lasalocid EC₅₀ ranged from 4 to 10 µM and microphotographs showed significant morphological changes of the cells after 24 h exposure. Silibinin EC₅₀ ranged from 40 to 42 µM for MTT and CBB assays, and 63 µM for LDH assay, and no significant morphological changes occurred. When lasalocid EC₅₀ was used in combination with silibinin in 1-250 µM concentrations, the EC₅₀ values were plotted at 36 µM and 72 µM in MTT and LDH assays, respectively. Thus co-actions of the two drugs led to significant diminishing of lasalocid cytotoxicity in respect to cellular metabolism and membrane integrity. The isobolograms showed remarkable antagonistic effect of silibinin in the course of lasalocid cytotoxic action. Although a considerable interaction was concisely relevant to hepatoma cell line FaO, the promising results incline to extend the study on other in vitro models (primary hepatocytes), as well as to investigate in vivo the cytoprotective effect of silibinin against lasalocid in target animals.
Opis fizyczny
  • Department of Pharmacology and Toxicology, National Veterinary Research Institute, 24-100 Pulawy, Poland
  • 1. Antonio R.V., Silva L.P., Vercesi A.E.: Alterations in mitochondrial Ca²⁺ flux by the antibiotic X-537A (lasalocid-A). Biochim Biophys Acta 1991, 1056, 250- 258.
  • 2. Avizeh R., Najafzadeh H., Razijalali M., Shirali S.: Evaluation of prophylactic and therapeutic effects of silymarin and N-acetylcysteine in acetaminophen-induced hepatotoxicity in cats. J Vet Pharmacol Ther 2009, 33, 95-99.
  • 3. Bergen W.G., Bates D.B.: Ionophores: their effect on production efficiency and mode of action. J Anim Sci 1984, 58, 1465-1483.
  • 4. Bolkent S., Zierold K.: Effects of the ionophores valinomycin, ionomycin and gramicidin A on the element compartmentation in cultured rat hepatocytes. Toxicol In Vitro 2002, 16, 159-165.
  • 5. Bradford M.M.: A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of dye binding. Anal Biochem 1976, 72, 248- 254.
  • 6. Chen C.H., Huang T.S., Wong C.H., Hong C.L., Tsai Y.H., Liang C.C., Lu F.J.: Synergistic anti-cancer effect of baicalein and silymarin on human hepatoma HepG2 Cells. Food Chem Toxicol 2009, 47, 638-644.
  • 7. Chou T.C., Talalay P.: Quantitative analysis of dose- effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regul 1984, 22, 27-55.
  • 8. Chou T.C., Talalay P.: Analysis of combined drug effects: A new look at a very old problem. Trends Pharmacol Sci 1983, 4, 450-454.
  • 9. Fraschini F., Demartini G., Esposti D.: Pharmacology of Silymarin. Clin Drug Investig 2002, 22, 51-65.
  • 10. Gazák R., Walterová D., Kren V.: Silybin and silymarin—new and emerging applications in medicine. Curr Med Chem 2007, 14, 315-338.
  • 11. Gessner P.K.: Isobolographic analysis of interactions: an update on applications and utility. Toxicology 1995, 105, 161-179.
  • 12. Grijalba M.T., Andrade P.B., Meinicke A.R., Castilho R.F., Vercesi A.E., Schreier S.: Inhibition of membrane lipid peroxidation by a radical scavenging mechanism: a novel function for hydroxyl-containing ionophores. Free Radie Res 1998, 28, 301-318.
  • 13. Grinde B.: Effect of carboxylic ionophores on lysosomal protein degradation in rat hepatocytes. Exp Cell Res 1983, 149, 27-35.
  • 14. Hogan F.S., Krishnegowda N.K., Mikhailova M., Kahlenberg M.S.: Flavonoid, silibinin, inhibits proliferation and promotes cell-cycle arrest of human colon cancer. J Surg Res 2007, 143, 58-65.
  • 15. Husain S.R., Cillard J., Cillard P.: Silymarin, a naturally occurring polyphenolic antioxidant flavonoid, inhibits azoxymethane-induced colon carcinogenesis in male F344 rats. Int J Cancer 1987, 101, 461-468.
  • 16. Mitsuaki M., Otake N.: Studies on the ionophorous antibiotics. XVI. The ionophore-mediated calcium transport and concomitant osmotic swelling of mitochondria. J Antibiot (Tokyo) 1978, 31, 888-893.
  • 17. Mollenhauer H.H., Morré D.J., Rowe L.D.: Alteration of intracellular traffic by monensin; mechanism, specificity and relationship to toxicity. Biochim Biophys Acta 1990, 1031, 225-246.
  • 18. Mosmann T.: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983, 65, 55-63.
  • 19. Novilla M.N.: The veterinary importance of the toxic syndrome induced by ionophores. Vet Hum Toxicol 1992, 34, 66-70.
  • 20. Olejnik M., Szprengier-Juszkiewicz T., Jedziniak P.: Confirmatory method for determination of coccidiostats in eggs. Bull Vet Inst Pulawy 2010, 54, 327-333.
  • 21. Radko L., Cybulski W., Rzeski W.: Cytotoxicity of salinomycin and lasalocid in a model hepatocyte cell line of a rat. "Medycyna Wet 2007, 63, 839-842.
  • 22. Ramadan L.A., Roushdy H.M., Abu Senna G.M., Amin N.E., El-Deshw O.A.: Radioprotective effect of silymarin against radiation induced hepatotoxicitx. Pharmacol Res 2002, 45, 447-454.
  • 23. Robak J., Gryglewski R.J.: Flavonoids are scavengers of superoxide anions. Biochem Pharmacol 1988, 37, 837- 841.
  • 24. Sasaki T., Kawai K., Saijo-Kurita K., Ohno T.: Detergent cytotoxicity: simplified assay of cytolysis by measuring LDH activity. Toxicol In Vitro 1992, 6, 451-457.
  • 25. Trost W., Halbach G.: Anti-phalloidine and anti-α- amanitine action of sily bin in comparison with compounds similar to structural parts of sily bin. Experientia 1978, 34, 1051-1052.
  • 26. Trinchet J.C., Coste T., Levy V.G.: Treatment of alcoholic hepatitis with silymarin. A double-blind comparative study in 116 patients. Gastroenterol Clin Biol 1989, 13, 120-124.
  • 27. Wellington K., Jarwis B.: Silymarin: a review of its clinical properties in the management of hepatic disorders. BioDrugs 2001, 15, 465-489.
Typ dokumentu
Identyfikator YADDA
JavaScript jest wyłączony w Twojej przeglądarce internetowej. Włącz go, a następnie odśwież stronę, aby móc w pełni z niej korzystać.