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

Respiratory Research

Hannu Kankaanranta¹  Vuokko L Kinnula²  Eeva Moilanen³  Pinja Ilmarinen-Salo³ 

[1] Department of Respiratory Medicine, Seinäjoki Central Hospital, Seinäjoki, Finland;Department of Medicine, Division of Pulmonary Medicine, University of Helsinki and Helsinki University Central Hospital, Helsinki, Finland;The Immunopharmacology Research Group, University of Tampere School of Medicine and Tampere University Hospital, Tampere, Finland

关键词: Reactive oxygen species;    JNK;    Mitochondrial permeability transition;    Nitric oxide;    Apoptosis;    Eosinophils;   

Others  :  796672
DOI  :  10.1186/1465-9921-13-73

 received in 2012-06-01, accepted in 2012-08-20,  发布年份 2012

【 摘 要 】

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.

【 授权许可】

   
2012 Ilmarinen-Salo et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

【 参考文献 】

• [1]Wenzel SE: Eosinophils in asthma–closing the loop or opening the door? N Engl J Med 2009, 360:1026-1028.
• [2]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.
• [3]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.
• [4]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.
• [5]Virchow JC Jr, 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.
• [6]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.
• [7]Korhonen R, Lahti A, Kankaanranta H, Moilanen E: Nitric oxide production and signaling in inflammation. Curr Drug Targets Inflamm Allergy 2005, 4:471-479.
• [8]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.
• [9]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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [16]Kroemer G, Galluzzi L, Brenner C: Mitochondrial membrane permeabilization in cell death. Physiol Rev 2007, 87:99-163.
• [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.
• [18]Dhanasekaran DN, Reddy EP: JNK signaling in apoptosis. Oncogene 2008, 27:6245-6251.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [27]Kinnula VL, Crapo JD: Superoxide dismutases in the lung and human lung diseases. Am J Respir Crit Care Med 2003, 167:1600-1619.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [35]Balakirev MY, Khramtsov VV, Zimmer G: Modulation of the mitochondrial permeability transition by nitric oxide. Eur J Biochem 1997, 246:710-718.
• [36]Cassarino DS, Halvorsen EM, Swerdlow RH, Abramova NN, Parker WD Jr, Sturgill TW, Bennett JP Jr: Interaction among mitochondria, mitogen-activated protein kinases, and nuclear factor-kappaB in cellular models of parkinson's disease. J Neurochem 2000, 74:1384-1392.
• [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.
• [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.
• [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.
• [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.
• [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.
• [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.
• [43]Zorov DB, Juhaszova M, Sollott SJ: Mitochondrial ROS-induced ROS release: An update and review. Biochim Biophys Acta 2006, 1757:509-517.
• [44]Green DR, Kroemer G: The pathophysiology of mitochondrial cell death. Science 2004, 305:626-629.
• [45]Kankaanranta H, Ilmarinen P, Zhang X, Nissinen E, Moilanen E: Antieosinophilic activity of orazipone. Mol Pharmacol 2006, 69:1861-1870.
• [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.
• [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.
• [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.
• [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.
• [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.