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. 1987 Nov;206(5):655–660. doi: 10.1097/00000658-198711000-00017

Alleviation of cyclosporine nephrotoxicity with verapamil and ATP-MgCl2. Mitochondrial respiratory and calcium studies.

B E Sumpio 1, A E Baue 1, I H Chaudry 1
PMCID: PMC1493310  PMID: 3499878

Abstract

Although recent studies have shown that combined treatment with verapamil and ATP-MgCl2 (ATP) prevents cyclosporine (CyA)-induced nephrotoxicity, the mechanism of these effects remains unknown. To study this, rat kidneys were perfused at 100 mmHg for 100 minutes with Krebs buffer containing 7.5 g/dL of albumin and substrates. After an equilibration period of 30 minutes, 500 ng/mL CyA was added. In some experiments 1 microgram/mL verapamil was added 10 minutes prior to CyA and in others 2 mM ATP was added to CyA. At the end of the perfusion, cortical mitochondria (mito) were isolated and mito Ca2+ and Mg2+ (mumoles/g protein) and respiratory control ratios (RCR) were measured. In addition, total tissue Ca2+ and Mg2+ levels were measured. The results indicate that CyA treatment leads to an accumulation of mito Ca2+ and a decrease in ADP/O ratio. Simultaneous administration of ATP with CyA led to an increased mito Ca2+ accumulation and depressed RCR, which were corrected by verapamil pretreatment. The combination of verapamil pretreatment and ATP cotreatment with CyA increased tissue ATP levels from 0.8 +/- 0.4 (control) to 1.4 +/- 0.1 mumol/g. This pharmacologic regimen may prevent CyA-induced nephrotoxicity by preventing mito Ca2+ accumulation and by preserving mitochondrial respiratory function. This allows a more efficient generation of ATP and consequently preservation of renal function.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  2. Carafoli E., Tiozzo R., Pasquali-Ronchetti I., Laschi R. A study of Ca 2+ metabolism in kidney mitochondria during acute uranium intoxication. Lab Invest. 1971 Dec;25(6):516–527. [PubMed] [Google Scholar]
  3. Chaudry I. H., Ohkawa M., Clemens M. G. Improved mitochondrial function following ischemia and reflow by ATP-MgCl2. Am J Physiol. 1984 May;246(5 Pt 2):R799–R804. doi: 10.1152/ajpregu.1984.246.5.R799. [DOI] [PubMed] [Google Scholar]
  4. GREENAWALT J. W., ROSSI C. S., LEHNINGER A. L. EFFECT OF ACTIVE ACCUMULATION OF CALCIUM AND PHOSPHATE IONS ON THE STRUCTURE OF RAT LIVER MITOCHONDRIA. J Cell Biol. 1964 Oct;23:21–38. doi: 10.1083/jcb.23.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Giant mitochondria, renal transplant biopsy, and cyclosporin A. Lancet. 1981 Jul 18;2(8238):146–146. [PubMed] [Google Scholar]
  6. Hayashi H., Chaudry I. H., Clemens M. G., Hull M. J., Baue A. E. Reoxygenation injury in isolated hepatocytes: effect of extracellular ATP on cation homeostasis. Am J Physiol. 1986 Apr;250(4 Pt 2):R573–R579. doi: 10.1152/ajpregu.1986.250.4.R573. [DOI] [PubMed] [Google Scholar]
  7. Humes H. D. Role of calcium in pathogenesis of acute renal failure. Am J Physiol. 1986 Apr;250(4 Pt 2):F579–F589. doi: 10.1152/ajprenal.1986.250.4.F579. [DOI] [PubMed] [Google Scholar]
  8. Maack T. Renal clearance and isolated kidney perfusion techniques. Kidney Int. 1986 Aug;30(2):142–151. doi: 10.1038/ki.1986.166. [DOI] [PubMed] [Google Scholar]
  9. McMillen M. A., Lewis T., Jaffe B. M., Wait R. B. Verapamil inhibition of lymphocyte proliferation and function in vitro. J Surg Res. 1985 Jul;39(1):76–80. doi: 10.1016/0022-4804(85)90164-7. [DOI] [PubMed] [Google Scholar]
  10. O'Donnell D., Rumbak M., Anderson J. Emphysematous pyelonephritis in a transplanted kidney. Clin Nephrol. 1986 Jan;25(1):52–53. [PubMed] [Google Scholar]
  11. Sumpio B. E., Chaudry I. H., Baue A. E. Adenosine triphosphate--magnesium chloride ameliorates reperfusion injury following ischemia as determined by phosphorus nuclear magnetic resonance. Arch Surg. 1985 Feb;120(2):233–240. doi: 10.1001/archsurg.1985.01390260091013. [DOI] [PubMed] [Google Scholar]
  12. Sumpio B. E., Chaudry I. H., Baue A. E. Reduction of the drug-induced nephrotoxicity by ATP-MgCl2. 1. Effects on the cis-diamminedichloroplatinum-treated isolated perfused kidneys. J Surg Res. 1985 May;38(5):429–437. doi: 10.1016/0022-4804(85)90058-7. [DOI] [PubMed] [Google Scholar]
  13. Sumpio B. E., Chaudry I. H., Baue A. E. Reduction of the drug-induced nephrotoxicity by ATP-MgCl2. II. Effects on gentamicin-treated isolated perfused kidneys. J Surg Res. 1985 May;38(5):438–445. doi: 10.1016/0022-4804(85)90059-9. [DOI] [PubMed] [Google Scholar]
  14. Sumpio B. E., Chaudry I. H., Clemens M. G., Baue A. E. Accelerated functional recovery of isolated rat kidney with ATP-MgCl2 after warm ischemia. Am J Physiol. 1984 Dec;247(6 Pt 2):R1047–R1053. doi: 10.1152/ajpregu.1984.247.6.R1047. [DOI] [PubMed] [Google Scholar]
  15. Sumpio B. E., Maack T. Kinetics, competition, and selectivity of tubular absorption of proteins. Am J Physiol. 1982 Oct;243(4):F379–F392. doi: 10.1152/ajprenal.1982.243.4.F379. [DOI] [PubMed] [Google Scholar]
  16. Sumpio B., Baue A. E., Chaudry I. H. Treatment with verapamil and adenosine triphosphate-MgCl2 reduces cyclosporine nephrotoxicity. Surgery. 1987 Mar;101(3):315–322. [PubMed] [Google Scholar]
  17. Weinberg J. M., Harding P. G., Humes H. D. Alterations in renal cortex cation homeostasis during mercuric chloride and gentamicin nephrotoxicity. Exp Mol Pathol. 1983 Aug;39(1):43–60. doi: 10.1016/0014-4800(83)90040-0. [DOI] [PubMed] [Google Scholar]
  18. Weinberg J. M., Harding P. G., Humes H. D. Mitochondrial bioenergetics during the initiation of mercuric chloride-induced renal injury. II. Functional alterations of renal cortical mitochondria isolated after mercuric chloride treatment. J Biol Chem. 1982 Jan 10;257(1):68–74. [PubMed] [Google Scholar]
  19. Whiting P. H., Thomson A. W., Blair J. T., Simpson J. G. Experimental cyclosporin A nephrotoxicity. Br J Exp Pathol. 1982 Feb;63(1):88–94. [PMC free article] [PubMed] [Google Scholar]
  20. Wilson D. R., Arnold P. E., Burke T. J., Schrier R. W. Mitochondrial calcium accumulation and respiration in ischemic acute renal failure in the rat. Kidney Int. 1984 Mar;25(3):519–526. doi: 10.1038/ki.1984.48. [DOI] [PubMed] [Google Scholar]
  21. Zoccarato F., Rugolo M., Siliprandi D., Siliprandi N. Correlated effluxes of adenine nucleotides, Mg2+ and Ca2+ induced in rat-liver mitochondria by external Ca2+ and phosphate. Eur J Biochem. 1981 Feb;114(2):195–199. doi: 10.1111/j.1432-1033.1981.tb05136.x. [DOI] [PubMed] [Google Scholar]

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