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Hyperglycemia induces apoptosis and p53 mobilization to mitochondria in RINm5F cells

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Abstract

The mechanisms related to hyperglycemia-induced pancreatic β-cell apoptosis are poorly defined. Rat insulin-producing cells (RINm5F) cultured in high glucose concentrations (30 mM) showed increased apoptosis and protein p53 translocation to mitochondria. In addition, hyperglycemia induced both the disruption of mitochondrial membrane potential (Δ < eqid1 > m), and an increase in reactive oxygen species (ROS), as shown by fluorescence changes of JC-1 and dichlorodihydrofluorescein-diacetate (DCDHF-DA), respectively. The increased intracellular ROS by high glucose exposure was blunted by mitochondrial-function and NADPH-oxidase inhibitors. We postulate that the concomitant mobilization of p53 protein to the mitochondria and the subsequent changes on the Δ < eqid2 > m, lead to an important pancreatic β-cell apoptosis mechanism induced by oxidative stress caused by hyperglycemia.

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References

  1. Burns TF, EL-Deiry WS: The p53 pathway and apoptosis. J Cell Physiol 181: 231–239, 1999

    Article  PubMed  CAS  Google Scholar 

  2. Brady HJ, Gil-Gómez G: The pro-apoptotic Bcl2 family member, Bax. Int J Biochem Cell Biol 30: 647–650, 1998

    Article  PubMed  CAS  Google Scholar 

  3. Sansome C, Zaika A, Marchenko ND, Moll UM: Hypoxia death stimulus induces translocation of p53 protein to mitochondria. Detection by immunofluorescence on whole cells. FEBS Lett 488: 110–115, 2001

    Article  PubMed  CAS  Google Scholar 

  4. Marchenko DN, Zaika A, Moll MU: Death signal-induced localization of p53 protein to mitochondria. A potential role in apoptotic signaling. J Biol Chem 275: 16202–16212, 2000

    Article  PubMed  CAS  Google Scholar 

  5. Macip S, Igarashi M, Berggren P, Yu J, Lee WS, Aaronson AS: Influence of induced reactive oxygen species in p53-mediated cell fate decisions. Mol Cell Biol 23: 8576–8585, 2003

    Article  PubMed  CAS  Google Scholar 

  6. Schuler M, Bossy-Wetzel E, Goldstein CJ, Fitzgerald P, Green RD: p53 induces apoptosis by caspase activation through mitochondrial cytochrome c release. J Biol Chem 275: 7337–7342, 2000

    Article  PubMed  CAS  Google Scholar 

  7. Mihara M, Erster S, Zaika A, Oleski P, Chittenden T, Pancoska P, Moil MU: P53 has a direct apoptogenic role at the mitochondria. Mol Cell 11: 577–590, 2003

    Article  PubMed  CAS  Google Scholar 

  8. Donahue JR, Razmara M, Hoek BJ, Knudsen BT: Direct influence of the p53 tumor suppressor on mitochondrial biogenesis and function. FASEB J 15: 635–644, 2001

    Article  PubMed  CAS  Google Scholar 

  9. Li PF, Dietz R, von Harsdorf R: p53 regulates mitochondrial membrane potential through reactive oxygen species and induces cytochrome c-independent apoptosis blocked by Bcl-2. EMBO J 18: 6027–6036, 1999

    PubMed  CAS  Google Scholar 

  10. Ito K, Nakazato T, Yamato K, Miyakawa Y, Yamada T, Hozumi N, Segawa K, Ikeda Y, Kizaki M: Induction of apoptosis in Leukemia cells by homovanillic acid derivate, capsaicin, through oxidative stress: implication of phosphorylation of p53 at Ser-15 residue by reactive oxygen species. Cancer Res 64: 1071–1078, 2004

    PubMed  CAS  Google Scholar 

  11. Bonini P, Cicconi S, Cardinales A, Vitale C, Serafino AL, Ciotti MT, Marlier NJ-LL: Oxidative stress induced p53-mediated apoptosis in glia: p53 transcription-independent way to die. J. Neurosc Res 75: 83–95, 2004

    CAS  Google Scholar 

  12. Donath YM, Gross JD, Cerasi KN: Hyperglycemia-induced β-cell apoptosis in pancreatic islets of Psammomys obesus during development of diabetes. Diabetes 48: 738–744, 1999

    PubMed  CAS  Google Scholar 

  13. Mandrup-Poulosen T: β-cell apoptosis. Diabetes 50 (suppl. 1): S58–S63, 2001

  14. Evans JL, Goldfine DI, Maddux AB, Grodsky MG: Oxidative stress and stress-activated signaling pathways: A unifying hypothesis of type 2 diabetes. Endocrine Rev 23: 599–622, 2002

    Article  CAS  Google Scholar 

  15. Marshak S, Leibowitz G, Bertzzi F, Kaiser N, Gross JD, Cerasi E, Melloul D: Impaired β-cell functions induced by chronic exposure of cultured human pancreatic islets to high glucose. Diabetes 48: 1230–1236, 1999

    PubMed  CAS  Google Scholar 

  16. Kaneto H, Kajimoto Y, Miyagawa J, Matsuoka T, Fujitani Y, Umayahara Y, Hanafusa T, Matsuzawa Y, Yamasaki Y, Hori M: Benefical effects of antioxidants in diabetes. Possible protection of pancreatic B-cells against glucose toxicity. Diabetes 48: 2398–2406,1999

  17. Brownlee M: Biochemistry and molecular cell biology of diabetic complications. Nature 414: 813–819, 2001

    Article  PubMed  CAS  Google Scholar 

  18. Sakai K, Matsumoto K, Nishikawa T, Suefuji M, Shirotani T, Brownlee M, Araki E: Mitochondrial reactive oxygen species reduce insulin secretion by pancreatic β cells. Biochem Biophys Res Comm 300: 216–222, 2003

    Article  PubMed  CAS  Google Scholar 

  19. Grankvist K, Marklund SL, Taljedal IB: CuZn-superoxide dismutase, Mn-superoxide dismutase, catalase and glutathione peroxidase in pancreatic islets and other tissues in the mouse. Biochem J 199: 393–398, 1981

    PubMed  CAS  Google Scholar 

  20. Navarro P, Valverde MA, Benito M, Lorenzo M: Insuline/IGF-I rescues immortalized brown adipocytes from apoptosis down regulating BclxL expression, in a PI 3-kinase- and Map kinase-dependent manner. Exp Cell Res 243: 213–221, 1998

    Article  PubMed  CAS  Google Scholar 

  21. Vrablic SA, Albrigth DC, Craciunescu NC, Salganik IR, Zeisel HS: Altered mitochondrial function and overgeneration of reactive oxygen species precede the induction of apoptosis by 1-O-octadecyl-2-methyl-rac-glycero-3-phosphocholine in p53-defective hepatocytes. FASEB J 15: 1739–1744, 2001

    Article  PubMed  CAS  Google Scholar 

  22. Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254, 1976

    Article  PubMed  CAS  Google Scholar 

  23. Bergmeyer HU: Methods of enzymatic analysis. Academic Press, New York, 1963, pp. 384–388

    Google Scholar 

  24. Fiordaliso F, Leri A, Cesselli D, Limana F, Safai B, Nadal-Ginard B, Anversa P, Kajstura J: Hyperglycemia activates p53 and p53-regulated genes leading to myocyte cell death. Diabetes 50: 2363–2375, 2001

    PubMed  CAS  Google Scholar 

  25. Kiningham KK, Oberley TD, Lin SM, Mattingly CA, Clair STKD: Overexpression of manganese superoxide dismutase protects against mitochondrial-initiated poly(ADP-ribose)polymerase-mediated cell death. FASEB J 13: 1601–1610, 1999

    PubMed  CAS  Google Scholar 

  26. Yamagishi SI, Edelstein D, Du LX, Brownlee M: Hyperglycemia potentiates collagen-induced platelet activation through mitochondrial superoxide overproduction. Diabetes 50: 1491–1494, 2001

    PubMed  CAS  Google Scholar 

  27. Meyer WJ, Schmitt EM: A central role of the endothelial NADPH oxidase in atherosclerosis. FEBS Lett 472: 1–4, 2000

    Article  PubMed  CAS  Google Scholar 

  28. Mathur A, Hong Y, Kemp BK, Barrientos AA, Erusalimsky JD: Evaluation of fluorescent dyes for the detection of mitochondrial membrane potential changes in cultured cardiomyocytes. Cardiov Res 46: 126–138, 2000

    Article  CAS  Google Scholar 

  29. Shiratsuchi A, Osada S, Kanazawa S, Nakanishi Y: Essential role of phosphatidylserine externalization in apoptosing cell phagocytosis by macrophages. Biochem Biophys Res Commun 246: 549–555, 1998

    Article  PubMed  CAS  Google Scholar 

  30. Gottlieb AR: Mitochondria: execution central. FEBS Lett 482: 6–12, 2000

    Article  PubMed  CAS  Google Scholar 

  31. Moll MU, Zaika A: Nuclear and mitochondrial apoptotic pathways of p53. FEBS Lett 493: 65–69, 2001

    Article  PubMed  CAS  Google Scholar 

  32. Chipuk JE, Kuwana T, Bouchier-Hayes L, Droin MN, Newmeyer DD, Schuler M, Green RD: Direct activation of Bax by p53 mediates mitochondrial membrane permeabilization and apoptosis. Science 303: 1010–1014, 2004

    Article  PubMed  CAS  Google Scholar 

  33. Chen QM, Bartholomew JC, Campisi J, Acosta M, Reagan JD, Ames BN: Molecular analysis of H2O2 induced senescent like growth arrest in normal human fibroblast: p53 and Rb control G1 arrest but not cell replication. Biochem J 332: 43–50, 1998

    PubMed  CAS  Google Scholar 

  34. Hussain SP, Amstad P, He P, Robles A, Lupold S, Kaneko I, Ichimiya M, Sengupta S, Mechanic L, Okamura S, Hofseth LJ, Moake M, Nagashima M, Forrester KS, Harris CC: p53-induced up-regulation of MnSOD and GPx but not Catalase increases oxidative stress and apoptosis. Cancer Res 64: 2350–2356, 2004

    PubMed  CAS  Google Scholar 

  35. Polyak K, Xia Y, Zweier LJ, Kinzler WK, Vogelstein B: A model for p53-induced apoptosis. Nature 389: 300–3005, 1997

    Article  PubMed  CAS  Google Scholar 

  36. Robertson PR, Harmon J, Tran OP, Tanaka Y, Takahashi H: Glucose toxicity in β-cells: type 2 diabetes, good radicals gone bad, and glutathione connection. Diabetes 52: 581–587, 2003

    PubMed  CAS  Google Scholar 

  37. Nishikawa T, Edelstein D, Du X-L, Yamagishi S, Matsumura T, Kaneda Y, Yorek MA, Beebe D, Oates JP, Hemmes HP, Giardino I, Brownlee M: Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature 404: 787–790, 2000

    PubMed  CAS  Google Scholar 

  38. Du X-L, Edelstein D, Rossetti L, Fantus IG, Goldberg H, Ziyadeh F, Wu J, Brownlee M: Hyperglycemia-induced mitochondrial superoxide overproduction activates the hexosamine pathway and induces plasminogen activator inhibitor-1 expression by increasing Sp1 glycosylation. Proc Natl Acad Sci USA 97: 12222–12226, 2000

    PubMed  CAS  Google Scholar 

  39. Oliveira HR, Verlengia R, Carvalho ROC, Britto GRL, Carpinelli RA: Pancreatic β-cells express phagocyte-like NAD(P)H oxidase. Diabetes 52: 1457–1463, 2003

    PubMed  CAS  Google Scholar 

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Authors and Affiliations

  1. Laboratorio de Bioquímica Inorgánica, Instituto Nacional de Enfermedades Respiratorias, México

    A. M. Guzmán-Grenfell

  2. Unidad de Investigación Médica en Bioquímica, Hospital de Especialidades. CMNS XXI. Instituto Mexicano del Seguro Social, México

    C. Ortega-Camarillo, R. García-Macedo, G. Durán-Reyes, R. Medina-Navarro, M. Cruz, M. Díaz-Flores & J. Kumate

  3. Bioquímica de la Reproducción, Departamento de Producción Agrícola y Animal, Universidad Autónoma Metropolitana, Unidad Xochimilco, México

    A. M. Rosales-Torres & A. Ávalos-Rodríguez

  4. Unidad de Investigación Médica en Bioquímica, Coordinación de Investigación en Salud, Centro Médico Nacional Siglo XXI, Instituto Mexicano del Seguro Social, Av. Cuauhtemoc 330, C.P. 06720, México D.F., México

    C. Ortega-Camarillo

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  1. C. Ortega-Camarillo
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  2. A. M. Guzmán-Grenfell
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Correspondence to C. Ortega-Camarillo.

Additional information

This work is part of the thesis required for the doctorate degree in Biological Sciences at the Universidad Autónoma Metropolitana, Mexico City, Mexico.

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Ortega-Camarillo, C., Guzmán-Grenfell, A.M., García-Macedo, R. et al. Hyperglycemia induces apoptosis and p53 mobilization to mitochondria in RINm5F cells. Mol Cell Biochem 281, 163–171 (2006). https://doi.org/10.1007/s11010-006-0829-5

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  • DOI: https://doi.org/10.1007/s11010-006-0829-5

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