Advertisement

Molecular and Cellular Biochemistry

, Volume 366, Issue 1–2, pp 335–343 | Cite as

Phytanic acid disturbs mitochondrial homeostasis in heart of young rats: a possible pathomechanism of cardiomyopathy in Refsum disease

  • Mateus Grings
  • Anelise Miotti Tonin
  • Lisiane Aurélio Knebel
  • Ângela Zanatta
  • Alana Pimentel Moura
  • Carlos Severo Dutra Filho
  • Moacir Wajner
  • Guilhian Leipnitz
Article

Abstract

Phytanic acid (Phyt) accumulates in tissues and biological fluids of patients affected by Refsum disease. Although cardiomyopathy is an important clinical manifestation of this disorder, the mechanisms of heart damage are poorly known. In the present study, we investigated the in vitro effects of Phyt on important parameters of oxidative stress in heart of young rats. Phyt significantly increased thiobarbituric acid-reactive substances levels (P < 0.001) and carbonyl formation (P < 0.01), indicating that this fatty acid induces lipid and protein oxidative damage, respectively. In contrast, Phyt did not alter sulfhydryl oxidation. Phyt also decreased glutathione (GSH) concentrations (P < 0.05), an important non-enzymatic antioxidant defense. Moreover, Phyt increased 2′,7′-dichlorofluorescin oxidation (DCFH) (P < 0.01), reflecting increased reactive species generation. We also found that the induced lipid and protein oxidative damage, as well as the decreased GSH levels and increased DCFH oxidation provoked by this fatty acid were prevented or attenuated by the reactive oxygen species scavengers melatonin, trolox, and GSH, but not by the nitric oxide inhibitor n ω-nitro-l-arginine methyl ester, suggesting that reactive oxygen species were involved in these effects. Next, we verified that Phyt strongly inhibited NADH-cytochrome c oxidoreductase (complex I–III) activity (P < 0.001) in heart supernatants, and decreased membrane potential and the NAD(P)H pool in heart mitochondria, indicating that Phyt acts as a metabolic inhibitor and as an uncoupler of the electron transport chain. Therefore, it can be presumed that disturbance of cellular energy and redox homeostasis induced by Phyt may possibly contribute to the cardiomyopathy found in patients affected by Refsum disease.

Keywords

Refsum disease Phytanic acid Oxidative stress Heart 

Notes

Acknowledgments

This work was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Programa de Apoio a Núcleos de Excelência (PRONEX II), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS), Pró-reitoria de Pesquisa/Universidade Federal do Rio Grande do Sul (PROPESQ/UFRGS), Financiadora de estudos e projetos (FINEP), Rede Instituto Brasileiro de Neurociência (IBN-Net) # 01.06.0842-00 and Instituto Nacional de Ciência e Tecnologia em Excitotoxicidade e Neuroproteção (INCT-EN).

References

  1. 1.
    Verhoeven NM, Jakobs C (2001) Human metabolism of phytanic and pristanic acid. Progr Lipid Res 40:453–466. doi: 10.1016/S0163-7827(01)00011-X CrossRefGoogle Scholar
  2. 2.
    Brosius U, Gärtner J (2002) Cellular and molecular aspects of Zellweger syndrome and other peroxisome biogenesis disorders. Cell Mol Life Sci 59:1058–1069PubMedCrossRefGoogle Scholar
  3. 3.
    Gould SJ, Raymond GV, Valle D (2001) The peroxisome biogenesis disorders. In: Scriver CR, Beaudet AL, Valle D (eds) The metabolic and molecular bases of inherited disease. McGraw-Hill, New York, pp 3181–3217Google Scholar
  4. 4.
    Wanders RJ, Schutgens RB, Barth PG, Tager JM, van den Bosch H (1993) Postnatal diagnosis of peroxisomal disorders: a biochemical approach. Biochimie 75:269–279. doi: 10.1016/0300-9084(93)90087-9 PubMedCrossRefGoogle Scholar
  5. 5.
    Al-Dirbashi OY, Santa T, Rashed MS, Al-Hassnan Z, Shimozawa N, Chedrawi A, Jacob M, Al-Mokhadab M (2008) Rapid UPLC-MS/MS method for routine analysis of plasma pristanic, phytanic, and very long chain fatty acid markers of peroxisomal disorders. J Lipid Res 49(8):1855–1862. doi: 10.1194/jlr.D800019-JLR200 PubMedCrossRefGoogle Scholar
  6. 6.
    Mönnig G, Wiekowski J, Kirchhof P, Stypmann J, Plenz G, Fabritz L, Bruns HJ, Eckardt L, Assmann G, Haverkamp W, Breithardt G, Seedorf U (2004) Phytanic acid accumulation is associated with conduction delay and sudden cardiac death in sterol carrier protein-2/sterol carrier protein-x deficient mice. J Cardiovasc Electrophysiol 15(11):1310–1316. doi: 10.1046/j.1540-8167.2004.03679.x PubMedCrossRefGoogle Scholar
  7. 7.
    Wanders RJA, Waterham HR, Leroy BP (2006) Refsum disease. In: Pagon RA, Bird TD, Dolan CR, Stephens K (eds) GeneReviews [Internet]. University of Washington, SeattleGoogle Scholar
  8. 8.
    Wanders RJ, Jakobs C, Skjeldal OH (2001) Refsum disease. In: Scriver CR, Beaudet AL, Sly WS, Valle D (eds) The metabolic and molecular bases of inherited disease. McGraw-Hill, New York, pp 3303–3322Google Scholar
  9. 9.
    Weinstein R (1999) Phytanic acid storage disease (Refsum’s disease): clinical characteristics, pathophysiology and the role of therapeutic apheresis in its management. J Clin Apher 14:181–184. doi: 10.1002/(SICI)1098-1101(1999)14:4<181:AID-JCA5>3.0.CO;2-Z PubMedCrossRefGoogle Scholar
  10. 10.
    Wierzbicki AS (2007) Peroxisomal disorders affecting phytanic acid α-oxidation: a review. Biochem Soc Trans 35(5):881–886PubMedCrossRefGoogle Scholar
  11. 11.
    Koh JT, Choi HH, Ahn KY, Kim JU, Kim JH, Chun JY, Baik YH, Kim KK (2001) Cardiac characteristics of transgenic mice overexpressing Refsum disease gene-associated protein within the heart. Biochem Biophys Res Commun 286:1107–1116. doi: 10.1006/bbrc.2001.5510 PubMedCrossRefGoogle Scholar
  12. 12.
    Baldwin EJ, Gibberd FB, Harley C, Sidey MC, Feher MD, Wierzbicki AS (2010) The effectiveness of long-term dietary therapy in the treatment of adult Refsum disease. J Neurol Neurosurg Psychiatry 81(9):954–957. doi: 10.1136/jnnp.2008.161059 PubMedCrossRefGoogle Scholar
  13. 13.
    Ferdinandusse S, Zomer AW, Komen JC, van den Brink CE, Thanos M, Hamers FP, Wanders RJ, van der Saag PT, Poll-The BT, Brites P (2008) Ataxia with loss of Purkinje cells in a mouse model for Refsum disease. Proc Natl Acad Sci USA 105:17712–17717. doi: 10.1073/pnas.0806066105 PubMedCrossRefGoogle Scholar
  14. 14.
    Gibberd FB, Billimoria JD, Page NG, Retsas S (1979) Heredopathia atactica polyneuritiformis (Refsum’s disease) treated by diet and plasma-exchange. Lancet 1:575–578. doi: 10.1016/S0140-6736(79)91005-5 PubMedCrossRefGoogle Scholar
  15. 15.
    Hungerbuhler JP, Meier C, Rousselle L, Quadri P, Bogousslavsky J (1985) Refsum’s disease: management by diet and plasmapheresis. Eur Neurol 24:153–159PubMedCrossRefGoogle Scholar
  16. 16.
    Masters-Thomas A, Bailes J, Billimoria JD, Clemens ME, Gibberd FB, Page NG (1980) Heredopathia atactica polyneuritiformis (Refsum’s disease): 1. Clinical features and dietary management. J Hum Nutr 34:245–250PubMedGoogle Scholar
  17. 17.
    Gibberd FB, Billimoria JD, Goldman JM, Clemens ME, Evans R, Whitelaw MN, Retsas S, Sherratt RM (1985) Heredopathia atactica polyneuritiformis: Refsum’s disease. Acta Neurol Scand 72:1–17PubMedCrossRefGoogle Scholar
  18. 18.
    Kahlert S, Schonfeld P, Reiser G (2005) The Refsum disease marker phytanic acid, a branched chain fatty acid, affects Ca2+ homeostasis and mitochondria, and reduces cell viability in rat hippocampal astrocytes. Neurobiol Dis 18:110–118. doi: 10.1016/j.nbd.2004.08.010 PubMedCrossRefGoogle Scholar
  19. 19.
    Komen JC, Distelmaier F, Koopman WJH, Wanders RJA, Smeitink J, Willems PHMG (2007) Phytanic acid impairs mitochondrial respiration through protonophoric action. Cell Mol Life Sci 64:3271–3281. doi: 10.1007/s00018-007-7357-7 PubMedCrossRefGoogle Scholar
  20. 20.
    Reiser G, Schonfeld P, Kahert S (2006) Mechanism of toxicity of the branched-chain fatty acid phytanic acid, a marker of Refsum disease, in astrocytes involves mitochondrial impairment. Int J Dev Neurosci 24:113–122. doi: 10.1016/j.ijdevneu.2005.11.002 PubMedCrossRefGoogle Scholar
  21. 21.
    Schönfeld P, Kahlert S, Reiser G (2004) In brain mitochondria the branched-chain fatty acid phytanic acid impairs energy transduction and sensitizes for permeability transition. Biochem J 383:121–128. doi: 10.1042/BJ20040583 PubMedCrossRefGoogle Scholar
  22. 22.
    Busanello EN, Viegas CM, Moura AP, Tonin AM, Grings M, Vargas CR, Wajner M (2010) In vitro evidence that phytanic acid compromises Na(+),K(+)-ATPase activity and the electron flow through the respiratory chain in brain cortex from young rats. Brain Res 1352:231–238. doi: 10.1016/j.brainres.2010.07.012 PubMedCrossRefGoogle Scholar
  23. 23.
    Leipnitz G, Amaral AU, Zanatta A, Seminotti B, Fernandes CG, Knebel LA, Vargas CR, Wajner M (2010) Neurochemical evidence that phytanic acid induces oxidative damage and reduces the antioxidant defenses in cerebellum and cerebral cortex of rats. Life Sci 87(9–10):275–280. doi: 10.1016/j.lfs.2010.06.015 PubMedCrossRefGoogle Scholar
  24. 24.
    Rönicke S, Kruska N, Kahlert S, Reiser G (2009) The influence of the branched-chain fatty acids pristanic acid and Refsum disease-associated phytanic acid on mitochondrial functions and calcium regulation of hippocampal neurons, astrocytes, and oligodendrocytes. Neurobiol Dis 36(2):401–410. doi: 10.1016/j.nbd.2009.08.005 PubMedCrossRefGoogle Scholar
  25. 25.
    Evelson P, Travacio M, Repetto M, Escobar J, Llesuy S, Lissi E (2001) Evaluation of total reactive antioxidant potential (TRAP) of tissue homogenates and their cytosols. Arch Biochem Biophys 388:261–266. doi: 10.1006/abbi.2001.2292 PubMedCrossRefGoogle Scholar
  26. 26.
    Ferranti R, da Silva MM, Kowaltowski AJ (2003) Mitochondrial ATP-sensitive K+ channel opening decreases reactive oxygen species generation. FEBS Lett 11(536):51–55. doi: 10.1016/S0014-5793(03)00007-3 CrossRefGoogle Scholar
  27. 27.
    Yagi K (1998) Simple procedure for specific assay of lipid hydroperoxides in serum or plasma. Methods Mol Biol 108:107–110PubMedGoogle Scholar
  28. 28.
    Reznick AZ, Packer L (1994) Oxidative damage to proteins: spectrophotometric method for carbonyl assay. Methods Enzymol 233:357–363PubMedCrossRefGoogle Scholar
  29. 29.
    Aksenov MY, Markesbery WR (2001) Change in thiol content and expression of glutathione redox system gene in the hippocampus and cerebellum in Alzheimer’s disease. Neurosci Lett 302:141–145. doi: 10.1016/S0304-3940(01)01636-6 PubMedCrossRefGoogle Scholar
  30. 30.
    Browne RW, Armstrong D (1998) Reduced glutathione and glutathione disulfide. Methods Mol Biol 108:347–352PubMedGoogle Scholar
  31. 31.
    Lebel CP, Ischiropoulos H, Bondy SC (1992) Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol 5:227–231PubMedCrossRefGoogle Scholar
  32. 32.
    Schapira AH, Mann VM, Cooper JM, Dexter D, Daniel S, Jenner P, Clark JB, Marsden CD (1990) Anatomic and disease specificity of NADH CoQ1 reductase (complex I) deficiency in Parkinson’s disease. J Neurochem 55:2142–2145. doi: 10.1111/j.1471-4159.1990.tb05809.x PubMedCrossRefGoogle Scholar
  33. 33.
    Akerman KE, Wikström MK (1976) Safranine as a probe of the mitochondrial membrane potential. FEBS Lett 68:191–197. doi: 10.1016/0014-5793(76)80434-6 PubMedCrossRefGoogle Scholar
  34. 34.
    Kowaltowski AJ, Cosso RG, Campos CB, Fiskum G (2002) Effect of Bcl-2 overexpression on mitochondrial structure and function. J Biol Chem 277:42802–42807. doi: 10.1074/jbc.M207765200 PubMedCrossRefGoogle Scholar
  35. 35.
    Maciel EN, Kowaltowski AJ, Schwalm FD, Rodrigues JM, Souza DO, Vercesi AE, Wajner M, Castilho RF (2004) Mitochondrial permeability transition in neuronal damage promoted by Ca21 and respiratory chain complex II inhibition. J Neurochem 90:1025–1035. doi: 10.1111/j.1471-4159.2004.02565.x PubMedCrossRefGoogle Scholar
  36. 36.
    Lowry OH, Rosebrough NJ, Lewis-Farr A, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  37. 37.
    Sugamura K, Keaney JF Jr (2011) Reactive oxygen species in cardiovascular disease. Free Radic Biol Med 51(5):978–992. doi: 10.1016/j.freeradbiomed.2011.05.004 PubMedCrossRefGoogle Scholar
  38. 38.
    Wattanapitayakul SK, Bauer JA (2001) Oxidative pathways in cardiovascular disease: roles, mechanisms, and therapeutic implications. Pharmacol Ther 89(2):187–206. doi: 10.1016/S0163-7258(00)00114-5 PubMedCrossRefGoogle Scholar
  39. 39.
    Halliwell B, Gutteridge JMC (2007) Measurement of reactive species. In: Halliwell B, Gutteridge JMC (eds) Free radicals in biology and medicine. Oxford University Press, Oxford, pp 268–340Google Scholar
  40. 40.
    Dalle-Donne I, Rossi R, Giustarini D, Milzani A, Colombo R (2003) Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta 329:23–38. doi: 10.1016/S0009-8981(03)00003-2 PubMedCrossRefGoogle Scholar
  41. 41.
    Levine RL (2002) Carbonyl modified proteins in cellular regulation, aging, and disease. Free Radic Biol Med 32:790–796. doi: 10.1016/S0891-5849(02)00765-7 PubMedCrossRefGoogle Scholar
  42. 42.
    Bilski P, Belanger AG, Chignell CF (2002) Photosensitized oxidation of 2′,7′-dichlorofluorescin: singlet oxygen does not contribute to the formation of fluorescent oxidation product 2′,7′-dichlorofluorescein. Free Radic Biol Med 33:938–946. doi: 10.1016/S0891-5849(02)00982-6 PubMedCrossRefGoogle Scholar
  43. 43.
    Ischiropoulos H, Gow A, Thom SR, Kooy NW, Royall JA, Crow JP (1999) Detection of reactive nitrogen species using 2,7-dichlorodihydrofluorescein and dihydrorhodamine 123. Methods Enzymol 301:367–373PubMedCrossRefGoogle Scholar
  44. 44.
    Myhre O, Andersen JM, Aarnes H, Fonnum F (2003) Evaluation of the probes 2′,7′-dichlorofluorescin diacetate, luminol, and lucigenin as indicators of reactive species formation. Biochem Pharmacol 65:1575–1582. doi: 10.1016/S0006-2952(03)00083-2 PubMedCrossRefGoogle Scholar
  45. 45.
    Ohashi T, Mizutani A, Murakami A, Kojo S, Ishii T, Taketani S (2002) Rapid oxidation of dichlorodihydrofluorescin with heme and hemoproteins: formation of the fluorescein is independent of the generation of reactive oxygen species. FEBS Lett 511:21–27. doi: 10.1016/S0014-5793(01)03262-8 PubMedCrossRefGoogle Scholar
  46. 46.
    Schönfeld P, Reiser G (2006) Rotenone-like action of the branched-chain phytanic acid induces oxidative stress in mitochondria. J Biol Chem 281(11):7136–7142. doi: 10.1074/jbc.M513198200 PubMedCrossRefGoogle Scholar
  47. 47.
    Schönfeld P, Reiser G (2008) Comment concerning the article: ‘Phytanic acid impairsmitochondrial respiration through protonophoric action’ by Komen et al.: branched chain phytanic acid inhibits the activity of the mitochondrial respiratory chain. Cell Mol Life Sci 65:2266–2269. doi: 10.1007/s00018-008-8117-z PubMedCrossRefGoogle Scholar
  48. 48.
    Wanders RJA, Janse GA, Lloyd MD (2003) Phytanic acid alpha-oxidation, new insights into an old problem: a review. Biochim Biophys Acta 1631:119–135. doi: 10.1016/S1388-1981(03)00003-9 PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2012

Authors and Affiliations

  • Mateus Grings
    • 1
  • Anelise Miotti Tonin
    • 1
  • Lisiane Aurélio Knebel
    • 1
  • Ângela Zanatta
    • 1
  • Alana Pimentel Moura
    • 1
  • Carlos Severo Dutra Filho
    • 1
  • Moacir Wajner
    • 1
    • 2
  • Guilhian Leipnitz
    • 1
  1. 1.Departamento de Bioquímica, Instituto de Ciências Básicas da SaúdeUniversidade Federal do Rio Grande do SulPorto AlegreBrazil
  2. 2.Serviço de Genética MédicaHospital de Clínicas de Porto AlegrePorto AlegreBrazil

Personalised recommendations