Top

Respiratory Research

Published in:

Open Access 01-12-2012 | Research

Nitric oxide-induced eosinophil apoptosis is dependent on mitochondrial permeability transition (mPT), JNK and oxidative stress: apoptosis is preceded but not mediated by early mPT-dependent JNK activation

Authors: Pinja Ilmarinen-Salo, Eeva Moilanen, Vuokko L Kinnula, Hannu Kankaanranta

Published in: Respiratory Research | Issue 1/2012

Abstract

Background

Eosinophils are critically involved in the pathogenesis of asthma. Nitric oxide (NO) is produced in high amounts in asthmatic lungs and has an important role as a regulator of lung inflammation. NO was previously shown to induce eosinophil apoptosis mediated via c-jun N-terminal kinase (JNK) and caspases. Our aim was to clarify the cascade of events leading to NO-induced apoptosis in granulocyte macrophage-colony stimulating factor (GM-CSF)-treated human eosinophils concentrating on the role of mitochondria, reactive oxygen species (ROS) and JNK.

Methods

Apoptosis was determined by flow cytometric analysis of relative DNA content, by Annexin-V labelling and/or morphological analysis. Immunoblotting was used to study phospho-JNK (pJNK) expression. Mitochondrial membrane potential was assessed by JC-1-staining and mitochondrial permeability transition (mPT) by loading cells with calcein acetoxymethyl ester (AM) and CoCl₂ after which flow cytometric analysis was conducted. Statistical significance was calculated by repeated measures analysis of variance (ANOVA) or paired t-test.

Results

NO-donor S-nitroso-N-acetyl-D,L-penicillamine (SNAP) induced late apoptosis in GM-CSF-treated eosinophils. SNAP-induced apoptosis was suppressed by inhibitor of mPT bongkrekic acid (BA), inhibitor of JNK SP600125 and superoxide dismutase-mimetic AEOL 10150. Treatment with SNAP led to late loss of mitochondrial membrane potential. Additionally, we found that SNAP induces early partial mPT (1 h) that was followed by a strong increase in pJNK levels (2 h). Both events were prevented by BA. However, these events were not related to apoptosis because SNAP-induced apoptosis was prevented as efficiently when BA was added 16 h after SNAP. In addition to the early and strong rise, pJNK levels were less prominently increased at 20–30 h.

Conclusions

Here we demonstrated that NO-induced eosinophil apoptosis is mediated via ROS, JNK and late mPT. Additionally, our results suggest that NO induces early transient mPT (flickerings) that leads to JNK activation but is not significant for apoptosis. Thereby, we showed some interesting early events in NO-stimulated eosinophils that may take place even if the threshold for irreversible mPT and apoptosis is not crossed. This study also revealed a previously unknown physiological function for transient mPT by showing that it may function as initiator of non-apoptotic JNK signalling.

Available only for authorised users

Wenzel SE: Eosinophils in asthma–closing the loop or opening the door?. N Engl J Med. 2009, 360: 1026-1028. 10.1056/NEJMe0900334.PubMedCrossRef

Akuthota P, Wang HB, Spencer LA, Weller PF: Immunoregulatory roles of eosinophils: A new look at a familiar cell. Clin Exp Allergy. 2008, 38: 1254-1263. 10.1111/j.1365-2222.2008.03037.x.PubMedPubMedCentralCrossRef

Humbles AA, Lloyd CM, McMillan SJ, Friend DS, Xanthou G, McKenna EE, Ghiran S, Gerard NP, Yu C, Orkin SH, Gerard C: A critical role for eosinophils in allergic airways remodeling. Science. 2004, 305: 1776-1779. 10.1126/science.1100283.PubMedCrossRef

Kankaanranta H, Lindsay MA, Giembycz MA, Zhang X, Moilanen E, Barnes PJ: Delayed eosinophil apoptosis in asthma. J Allergy Clin Immun. 2000, 106: 77-83. 10.1067/mai.2000.107038.PubMedCrossRef

Virchow JC, Walker C, Hafner D, Kortsik C, Werner P, Matthys H, Kroegel C: T cells and cytokines in bronchoalveolar lavage fluid after segmental allergen provocation in atopic asthma. Am J Respir Crit Care Med. 1995, 151: 960-968.PubMed

Park CS, Choi YS, Ki SY, Moon SH, Jeong SW, Uh ST, Kim YH: Granulocyte macrophage colony-stimulating factor is the main cytokine enhancing survival of eosinophils in asthmatic airways. Eur Respir J. 1998, 12: 872-878. 10.1183/09031936.98.12040872.PubMedCrossRef

Korhonen R, Lahti A, Kankaanranta H, Moilanen E: Nitric oxide production and signaling in inflammation. Curr Drug Targets Inflamm Allergy. 2005, 4: 471-479. 10.2174/1568010054526359.PubMedCrossRef

Lehtimaki L, Kankaanranta H, Saarelainen S, Hahtola P, Jarvenpaa R, Koivula T, Turjanmaa V, Moilanen E: Extended exhaled NO measurement differentiates between alveolar and bronchial inflammation. Am J Respir Crit Care Med. 2001, 163: 1557-1561.PubMedCrossRef

Payne DN, Adcock IM, Wilson NM, Oates T, Scallan M, Bush A: Relationship between exhaled nitric oxide and mucosal eosinophilic inflammation in children with difficult asthma, after treatment with oral prednisolone. Am J Respir Crit Care Med. 2001, 164: 1376-1381.PubMedCrossRef

10.

Oliveira MS, de O Barreto E, Zamuner S, Pires AL, Ferreira TP, Cordeiro RS, Lagente V, Martins MA, Wallace JL, e Silva PM: Suppressive effects of nitric oxide-releasing prednisolone NCX-1015 on the allergic pleural eosinophil recruitment in rats. Clin Exp Allergy. 2008, 38: 1830-1837.PubMed

11.

Zhang X, Moilanen E, Lahti A, Hamalainen M, Giembycz MA, Barnes PJ, Lindsay MA, Kankaanranta H: Regulation of eosinophil apoptosis by nitric oxide: Role of c-jun-N-terminal kinase and signal transducer and activator of transcription 5. J Allergy Clin Immun. 2003, 112: 93-101. 10.1067/mai.2003.1587.PubMedCrossRef

12.

Ilmarinen-Salo P, Moilanen E, Kankaanranta H: Nitric oxide induces apoptosis in GM-CSF-treated eosinophils via caspase-6-dependent lamin and DNA fragmentation. Pulm Pharmacol Ther. 2010, 23: 365-371. 10.1016/j.pupt.2010.04.001.PubMedCrossRef

13.

Pontin J, Blaylock MG, Walsh GM, Turner SW: Sputum eosinophil apoptotic rate is positively correlated to exhaled nitric oxide in children. Pediatr Pulmonol. 2008, 43: 1130-1134. 10.1002/ppul.20921.PubMedCrossRef

14.

Brown GC, Borutaite V: Nitric oxide inhibition of mitochondrial respiration and its role in cell death. Free Radic Biol Med. 2002, 33: 1440-1450. 10.1016/S0891-5849(02)01112-7.PubMedCrossRef

15.

Hortelano S, Dallaporta B, Zamzami N, Hirsch T, Susin SA, Marzo I, Bosca L, Kroemer G: Nitric oxide induces apoptosis via triggering mitochondrial permeability transition. FEBS Lett. 1997, 410: 373-377. 10.1016/S0014-5793(97)00623-6.PubMedCrossRef

16.

Kroemer G, Galluzzi L, Brenner C: Mitochondrial membrane permeabilization in cell death. Physiol Rev. 2007, 87: 99-163. 10.1152/physrev.00013.2006.PubMedCrossRef

17.

Letuve S, Druilhe A, Grandsaigne M, Aubier M, Pretolani M: Critical role of mitochondria, but not caspases, during glucocorticosteroid-induced human eosinophil apoptosis. Am J Respir Cell Mol Biol. 2002, 26: 565-571.PubMedCrossRef

18.

Dhanasekaran DN, Reddy EP: JNK signaling in apoptosis. Oncogene. 2008, 27: 6245-6251. 10.1038/onc.2008.301.PubMedPubMedCentralCrossRef

19.

Salvioli S, Ardizzoni A, Franceschi C, Cossarizza A: JC-1, but not DiOC6(3) or rhodamine 123, is a reliable fluorescent probe to assess delta psi changes in intact cells: Implications for studies on mitochondrial functionality during apoptosis. FEBS Lett. 1997, 411: 77-82. 10.1016/S0014-5793(97)00669-8.PubMedCrossRef

20.

Petronilli V, Miotto G, Canton M, Brini M, Colonna R, Bernardi P, Di Lisa F: Transient and long-lasting openings of the mitochondrial permeability transition pore can be monitored directly in intact cells by changes in mitochondrial calcein fluorescence. Biophys J. 1999, 76: 725-734. 10.1016/S0006-3495(99)77239-5.PubMedPubMedCentralCrossRef

21.

Petronilli V, Penzo D, Scorrano L, Bernardi P, Di Lisa F: The mitochondrial permeability transition, release of cytochrome c and cell death. correlation with the duration of pore openings in situ. J Biol Chem. 2001, 276: 12030-12034. 10.1074/jbc.M010604200.PubMedCrossRef

22.

Ichas F, Jouaville LS, Mazat JP: Mitochondria are excitable organelles capable of generating and conveying electrical and calcium signals. Cell. 1997, 89: 1145-1153. 10.1016/S0092-8674(00)80301-3.PubMedCrossRef

23.

Ventura JJ, Hubner A, Zhang C, Flavell RA, Shokat KM, Davis RJ: Chemical genetic analysis of the time course of signal transduction by JNK. Mol Cell. 2006, 21: 701-710. 10.1016/j.molcel.2006.01.018.PubMedCrossRef

24.

Sanchez-Perez I, Murguia JR, Perona R: Cisplatin induces a persistent activation of JNK that is related to cell death. Oncogene. 1998, 16: 533-540. 10.1038/sj.onc.1201578.PubMedCrossRef

25.

Hanawa N, Shinohara M, Saberi B, Gaarde WA, Han D, Kaplowitz N: Role of JNK translocation to mitochondria leading to inhibition of mitochondria bioenergetics in acetaminophen-induced liver injury. J Biol Chem. 2008, 283: 13565-13577. 10.1074/jbc.M708916200.PubMedPubMedCentralCrossRef

26.

Lin X, Wang YJ, Li Q, Hou YY, Hong MH, Cao YL, Chi ZQ, Liu JG: Chronic high-dose morphine treatment promotes SH-SY5Y cell apoptosis via c-jun N-terminal kinase-mediated activation of mitochondria-dependent pathway. FEBS J. 2009, 276: 2022-2036. 10.1111/j.1742-4658.2009.06938.x.PubMedCrossRef

27.

Kinnula VL, Crapo JD: Superoxide dismutases in the lung and human lung diseases. Am J Respir Crit Care Med. 2003, 167: 1600-1619. 10.1164/rccm.200212-1479SO.PubMedCrossRef

28.

Halestrap AP, Brenner C: The adenine nucleotide translocase: A central component of the mitochondrial permeability transition pore and key player in cell death. Curr Med Chem. 2003, 10: 1507-1525. 10.2174/0929867033457278.PubMedCrossRef

29.

Kokoszka JE, Waymire KG, Levy SE, Sligh JE, Cai J, Jones DP, MacGregor GR, Wallace DC: The ADP/ATP translocator is not essential for the mitochondrial permeability transition pore. Nature. 2004, 427: 461-465. 10.1038/nature02229.PubMedPubMedCentralCrossRef

30.

Vieira HL, Belzacq AS, Haouzi D, Bernassola F, Cohen I, Jacotot E, Ferri KF, El Hamel C, Bartle LM, Melino G, Brenner C, Goldmacher V, Kroemer G: The adenine nucleotide translocator: A target of nitric oxide, peroxynitrite, and 4-hydroxynonenal. Oncogene. 2001, 20: 4305-4316. 10.1038/sj.onc.1204575.PubMedCrossRef

31.

Ma Q, Fang H, Shang W, Liu L, Xu Z, Ye T, Wang X, Zheng M, Chen Q, Cheng H: Superoxide flashes: Early mitochondrial signals for oxidative stress-induced apoptosis. J Biol Chem. 2011, 286: 27573-27581. 10.1074/jbc.M111.241794.PubMedPubMedCentralCrossRef

32.

Bernardi P, Petronilli V: The permeability transition pore as a mitochondrial calcium release channel: A critical appraisal. J Bioenerg Biomembr. 1996, 28: 131-138. 10.1007/BF02110643.PubMedCrossRef

33.

Barsukova A, Komarov A, Hajnoczky G, Bernardi P, Bourdette D, Forte M: Activation of the mitochondrial permeability transition pore modulates Ca2+ responses to physiological stimuli in adult neurons. Eur J Neurosci. 2011, 33: 831-842. 10.1111/j.1460-9568.2010.07576.x.PubMedPubMedCentralCrossRef

34.

Saotome M, Katoh H, Yaguchi Y, Tanaka T, Urushida T, Satoh H, Hayashi H: Transient opening of mitochondrial permeability transition pore by reactive oxygen species protects myocardium from ischemia-reperfusion injury. Am J Physiol Heart Circ Physiol. 2009, 296: H1125-H1132. 10.1152/ajpheart.00436.2008.PubMedCrossRef

35.

Balakirev MY, Khramtsov VV, Zimmer G: Modulation of the mitochondrial permeability transition by nitric oxide. Eur J Biochem. 1997, 246: 710-718. 10.1111/j.1432-1033.1997.00710.x.PubMedCrossRef

36.

Cassarino DS, Halvorsen EM, Swerdlow RH, Abramova NN, Parker WD, Sturgill TW, Bennett JP: Interaction among mitochondria, mitogen-activated protein kinases, and nuclear factor-kappaB in cellular models of parkinson's disease. J Neurochem. 2000, 74: 1384-1392.PubMedCrossRef

37.

Beltran B, Mathur A, Duchen MR, Erusalimsky JD, Moncada S: The effect of nitric oxide on cell respiration: A key to understanding its role in cell survival or death. Proc Natl Acad Sci USA. 2000, 97: 14602-14607. 10.1073/pnas.97.26.14602.PubMedPubMedCentralCrossRef

38.

Peachman KK, Lyles DS, Bass DA: Mitochondria in eosinophils: Functional role in apoptosis but not respiration. Proc Natl Acad Sci USA. 2001, 98: 1717-1722. 10.1073/pnas.98.4.1717.PubMedPubMedCentralCrossRef

39.

Hebestreit H, Dibbert B, Balatti I, Braun D, Schapowal A, Blaser K, Simon HU: Disruption of fas receptor signaling by nitric oxide in eosinophils. J Exp Med. 1998, 187: 415-425. 10.1084/jem.187.3.415.PubMedPubMedCentralCrossRef

40.

Beauvais F, Michel L, Dubertret L: The nitric oxide donors, azide and hydroxylamine, inhibit the programmed cell death of cytokine-deprived human eosinophils. FEBS Lett. 1995, 361: 229-232. 10.1016/0014-5793(95)00188-F.PubMedCrossRef

41.

Beauvais F, Joly F: Effects of nitric oxide on the eosinophil survival in vitro. A role for nitrosyl-heme. FEBS Lett. 1999, 443: 37-40.PubMedCrossRef

42.

Gardai SJ, Hoontrakoon R, Goddard CD, Day BJ, Chang LY, Henson PM, Bratton DL: Oxidant-mediated mitochondrial injury in eosinophil apoptosis: Enhancement by glucocorticoids and inhibition by granulocyte-macrophage colony-stimulating factor. J Immunol. 2003, 170: 556-566.PubMedCrossRef

43.

Zorov DB, Juhaszova M, Sollott SJ: Mitochondrial ROS-induced ROS release: An update and review. Biochim Biophys Acta. 2006, 1757: 509-517. 10.1016/j.bbabio.2006.04.029.PubMedCrossRef

44.

Green DR, Kroemer G: The pathophysiology of mitochondrial cell death. Science. 2004, 305: 626-629. 10.1126/science.1099320.PubMedCrossRef

45.

Kankaanranta H, Ilmarinen P, Zhang X, Nissinen E, Moilanen E: Antieosinophilic activity of orazipone. Mol Pharmacol. 2006, 69: 1861-1870. 10.1124/mol.105.021170.PubMedCrossRef

46.

Morishima Y, Gotoh Y, Zieg J, Barrett T, Takano H, Flavell R, Davis RJ, Shirasaki Y, Greenberg ME: Beta-amyloid induces neuronal apoptosis via a mechanism that involves the c-jun N-terminal kinase pathway and the induction of fas ligand. J Neurosci. 2001, 21: 7551-7560.PubMed

47.

Cleeter MW, Cooper JM, Darley-Usmar VM, Moncada S, Schapira AH: Reversible inhibition of cytochrome c oxidase, the terminal enzyme of the mitochondrial respiratory chain, by nitric oxide. implications for neurodegenerative diseases. FEBS Lett. 1994, 345: 50-54. 10.1016/0014-5793(94)00424-2.PubMedCrossRef

48.

Wei T, Chen C, Hou J, Xin W, Mori A: Nitric oxide induces oxidative stress and apoptosis in neuronal cells. Biochim Biophys Acta. 2000, 1498: 72-79. 10.1016/S0167-4889(00)00078-1.PubMedCrossRef

49.

Machiavelli LI, Poliandri AH, Quinteros FA, Cabilla JP, Duvilanski BH: Reactive oxygen species are key mediators of the nitric oxide apoptotic pathway in anterior pituitary cells. Nitric Oxide. 2007, 16: 237-246. 10.1016/j.niox.2006.08.002.PubMedCrossRef

50.

Moriya R, Uehara T, Nomura Y: Mechanism of nitric oxide-induced apoptosis in human neuroblastoma SH-SY5Y cells. FEBS Lett. 2000, 484: 253-260. 10.1016/S0014-5793(00)02167-0.PubMedCrossRef

Title: Nitric oxide-induced eosinophil apoptosis is dependent on mitochondrial permeability transition (mPT), JNK and oxidative stress: apoptosis is preceded but not mediated by early mPT-dependent JNK activation
Authors: Pinja Ilmarinen-Salo
Eeva Moilanen
Vuokko L Kinnula
Hannu Kankaanranta
Publication date: 01-12-2012
Publisher: BioMed Central
Published in: Respiratory Research / Issue 1/2012
Electronic ISSN: 1465-993X
DOI: https://doi.org/10.1186/1465-9921-13-73

Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Reserve your place

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin

Prof. Florian Limbourg

Prof. Anoop Chauhan

Developed by: Springer Medicine

Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2012

Emphysema distribution and annual changes in pulmonary function in male patients with chronic obstructive pulmonary disease

Airway responses and inflammation in subjects with asthma after four days of repeated high-single-dose allergen challenge

Observational study of the outcomes and costs of initiating maintenance therapies in patients with moderate exacerbations of COPD

Measurement of asthma control according to global initiative for asthma guidelines: a comparison with the asthma control questionnaire

Differential effects of formoterol on thrombin- and PDGF-induced proliferation of human pulmonary arterial vascular smooth muscle cells

Mechanical ventilation with high tidal volumes attenuates myocardial dysfunction by decreasing cardiac edema in a rat model of LPS-induced peritonitis

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

At a glance: The STEP trials