vestnik-bio-msuВестник Московского университета. Серия 16. БиологияVestnik Moskovskogo universiteta. Seriya 16. Biologiya0137-0952Lomonosov Moscow State University, School of Biology10.55959/MSU0137-0952-16-78-2-4vestnik-bio-msu-1232Research ArticleОРИГИНАЛЬНОЕ ИССЛЕДОВАНИЕRESEARCH ARTICLEВлияние карбонилцианид-4-(трифторметокси)фенилгидразона (FCCP) на FcεRI-зависимую продукцию цитокинов клетками RBL-2H3Effect of 4-(trifluoromethoxy)phenylhydrazone carbonyl cyanide (FCCP) on FcεRI-dependent cytokine production by RBL-2H3 cellshttps://orcid.org/0000-0003-3694-8007ПавлюченковаА. Н.PavlyuchenkovaA. N.

Павлюченкова Анастасия Никитична – аспирантка  факультета биоинженерии и биоинформатики, инженер 1-й категории отд. мат. методов в биологии. Тел.: 8-495-939-03-38

119992, г. Москва, Ленинские горы, д. 1, стр. 40;

Факультет биоинженерии и биоинформатики

119992, г. Москва, Ленинские горы, д. 1, стр. 73

1–40 Leninskie gory, Moscow, 119992;

Faculty of Bioengineering and Bioinformatics

1–73 Leninskie gory, Moscow, 119992

anabella.gen@gmail.comhttps://orcid.org/0009-0009-8897-1517СмирновМ. С.SmirnovM. S.

Смирнов Максим Сергеевич – студент факультета биоинженерии и биоинформатики, ст. лаборант отд. мат. методов в биологии. Тел.: 8-495-939-03-38

119992, г. Москва, Ленинские горы, д. 1, стр. 40;

Факультет биоинженерии и биоинформатики

119992, г. Москва, Ленинские горы, д. 1, стр. 73

1–40 Leninskie gory, Moscow, 119992;

Faculty of Bioengineering and Bioinformatics

1–73 Leninskie gory, Moscow, 119992

maksimsmirnov1028@gmail.comhttps://orcid.org/0000-0002-3902-7812ЧеломбитькоМ. А.ChelombitkoM. A.

Челомбитько Мария Александровна – канд. биол. наук, ст. науч. сотр. отд. мат. методов в биологии. Тел.: 8-495-939-03-38

119992, г. Москва, Ленинские горы, д. 1, стр. 40

1–40 Leninskie gory, Moscow, 119992

chelombitko@mail.bio.msu.ruНаучно-исследовательский институт физико-химической биологии имени А.Н. Белозерского, Московский государственный университет имени М.В. Ломоносова; Московский государственный университет имени М.В. ЛомоносоваРоссияBelozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University; Lomonosov oscow State UniversityRussian FederationНаучно-исследовательский институт физико-химической биологии имени А.Н. Белозерского, Московский государственный университет имени М.В. ЛомоносоваРоссияBelozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State UniversityRussian Federation2023090720237827885Copyright © Павлюченкова А.Н., Смирнов М.С., Челомбитько М.А., 20232023Павлюченкова А.Н., Смирнов М.С., Челомбитько М.А.Pavlyuchenkova A.N., Smirnov M.S., Chelombitko M.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestnik-bio-msu.elpub.ru/jour/article/view/1232

Ключевое значение в развитии аллергических заболеваний имеют тучные клетки (ТК). Основную роль в активации ТК при аллергии играет взаимодействие антигенов с иммуноглобулином E и последующее связывание этих комплексов с рецептором FcεRI, что в конечном счете приводит к быстрому экзоцитозу гранул и последующей продукции цитокинов. Имеются данные о роли мембранного потенциала митохондрий в FcεRIзависимой активации ТК. Так, применение классических разобщителей окислительного фосфорилирования снижает уровень дегрануляции ТК. Однако их влияние на продукцию цитокинов ТК не изучено. В настоящей работе было продемонстрировано, что предварительная обработка ТК линии RBL-2H3 разобщителем карбонилцианид-4-(трифторметокси)фенилгидразоном (FCCP) приводит к уменьшению не только уровня FcεRI-зависимой дегрануляции, но и продукции цитокинов TNFα и IL-4. При этом FCCP препятствует фосфорилированию адаптерной молекулы LAT, а также киназы Erk1/2, что может лежать в основе ингибирующего влияния разобщителя на FcεRIзависимую активацию клеток линии RBL-2H3. Полученные данные свидетельствуют о том, что мембранный потенциал митохондрий играет важную роль в FcεRI-зависимой активации ТК, а разобщение окислительного фосфорилирования и дыхания митохондрий с помощью разобщителей может использоваться для регуляции этого процесса.

Mast cells (MCs) play a key role in the development of allergic diseases. The interaction of antigens with immunoglobulin E and the subsequent binding of these complexes to the FcεRI receptor, which ultimately leads to rapid exocytosis of granules and subsequent production of cytokines, play a major role in MC activation in allergy. There is data on the role of the mitochondrial membrane potential in the FcεRI-dependent activation of MC. Thus, the use of classical uncouplers of oxidative phosphorylation reduces MC degranulation. However, their effect on the production of MC cytokines has not been studied. In the present work, it was demonstrated that pretreatment of RBL-2H3 mast cells with the uncoupler carbonyl cyanide-4-(trifluoromethoxy)phenylhydrazone (FCCP) leads to a decrease not only in FcεRI-dependent degranulation, but also in a decrease in the production of TNFα and IL-4 cytokines. At the same time, FCCP prevents the phosphorylation of the LAT adapter molecule, as well as the Erk1/2 kinase, which may underlie the inhibitory effect of the uncoupler on FcεRI-dependent activation of RBL-2H3 cell line. The data obtained indicate that the mitochondrial membrane potential plays an important role in the FcεRI-dependent activation of MC, and the uncoupling of oxidative phosphorylation and respiration of mitochondria with the help of uncouplers can be used to regulate this process.

тучные клеткиразобщителимитохондрииаллергияFcεRI-зависимая активацияFCCPmast cellsuncouplersmitochondriaallergyFcεRI-dependent activationFCCPРабота выполнено при финансовой поддержке Российского научного фонда (проект № 22-74-00081).The research was supported by Russian Science Foundation (project No 22-74-00081).Referencesda Silva E.Z.M., Jamur M.C., Oliver C. Mast cell function: a new vision of an old cell. J. Histochem. Cytochem. 2014;62(10):698–738.da Silva E.Z.M., Jamur M.C., Oliver C. Mast cell function: a new vision of an old cell. J. Histochem. Cytochem. 2014;62(10):698–738.Krystel-Whittemore M., Dileepan K.N., Wood J.G. Mast cell: A multi-functional master cell. Front. Immunol. 2015;6:620.Krystel-Whittemore M., Dileepan K.N., Wood J.G. Mast cell: A multi-functional master cell. Front. Immunol. 2015;6:620.Wernersson S., Pejler G. Mast cell secretory granules: armed for battle. Nat. Rev. Immunol. 2014;14(7):478–494.Wernersson S., Pejler G. Mast cell secretory granules: armed for battle. Nat. Rev. Immunol. 2014;14(7):478–494.Pejler G., Hu Frisk J.M., Sjöström D., Paivandy A., Öhrvik H. Acidic pH is essential for maintaining mast cell secretory granule homeostasis. Cell Death Dis. 2017;8(5):e2785.Pejler G., Hu Frisk J.M., Sjöström D., Paivandy A., Öhrvik H. Acidic pH is essential for maintaining mast cell secretory granule homeostasis. Cell Death Dis. 2017;8(5):e2785.Kanagaratham C., El Ansari Y.S., Lewis O.L., Oettgen H.C. IgE and IgG antibodies as regulators of mast cell and basophil functions in food allergy. Front. Immunol. 2020;11:603050.Kanagaratham C., El Ansari Y.S., Lewis O.L., Oettgen H.C. IgE and IgG antibodies as regulators of mast cell and basophil functions in food allergy. Front. Immunol. 2020;11:603050.Mohr F. C., Fewtrell C. The relative contributions of extracellular and intracellular calcium to secretion from tumor mast cells. Multiple effects of the proton ionophore carbonyl cyanide m-chlorophenylhydrazone. J. Biol. Chem. 1987;262(22):10638–10643.Mohr F. C., Fewtrell C. The relative contributions of extracellular and intracellular calcium to secretion from tumor mast cells. Multiple effects of the proton ionophore carbonyl cyanide m-chlorophenylhydrazone. J. Biol. Chem. 1987;262(22):10638–10643.Suzuki Y., Yoshimaru T., Inoue T., Ra C. Mitochondrial Ca2+ flux is a critical determinant of the Ca2+ dependence of mast cell degranulation. J. Leukoc. Biol. 2006;79(3):508–518.Suzuki Y., Yoshimaru T., Inoue T., Ra C. Mitochondrial Ca2+ flux is a critical determinant of the Ca2+ dependence of mast cell degranulation. J. Leukoc. Biol. 2006;79(3):508–518.Weatherly L.M., Shim J., Hashmi H.N., Kennedy R.H., Hess S.T., Gosse J.A. Antimicrobial agent triclosan is a proton ionophore uncoupler of mitochondria in living rat and human mast cells and in primary human keratinocytes. J. Appl. Toxicol. 2016;36(6):777–789.Weatherly L.M., Shim J., Hashmi H.N., Kennedy R.H., Hess S.T., Gosse J.A. Antimicrobial agent triclosan is a proton ionophore uncoupler of mitochondria in living rat and human mast cells and in primary human keratinocytes. J. Appl. Toxicol. 2016;36(6):777–789.Weatherly L.M. Nelson A.J., Shim J., Riitano A.M., Gerson E.D., Hart A.J., de Juan-Sanz J., Ryan T.A., Sher R., Hess S.T., Gosse J.A. Antimicrobial agent triclosan disrupts mitochondrial structure, revealed by super-resolution microscopy, and inhibits mast cell signaling via calcium modulation. Toxicol. Appl. Pharmacol. 2018;349:39–54.Weatherly L.M. Nelson A.J., Shim J., Riitano A.M., Gerson E.D., Hart A.J., de Juan-Sanz J., Ryan T.A., Sher R., Hess S.T., Gosse J.A. Antimicrobial agent triclosan disrupts mitochondrial structure, revealed by super-resolution microscopy, and inhibits mast cell signaling via calcium modulation. Toxicol. Appl. Pharmacol. 2018;349:39–54.Demine S., Renard P., Arnould T. Mitochondrial uncoupling: A key controller of biological processes in physiology and diseases. Cells. 2019;8(8):795.Demine S., Renard P., Arnould T. Mitochondrial uncoupling: A key controller of biological processes in physiology and diseases. Cells. 2019;8(8):795.Sagi-Eisenberg R., Pecht I. Resolution of cellular compartments involved in membrane potential changes accompanying IgE-mediated degranulation of rat basophilic leukemia cells. EMBO J. 1984;3(3):497–500.Sagi-Eisenberg R., Pecht I. Resolution of cellular compartments involved in membrane potential changes accompanying IgE-mediated degranulation of rat basophilic leukemia cells. EMBO J. 1984;3(3):497–500.Inoue T., Suzuki Y., Ra C. Epigallocatechin-3-gallate inhibits mast cell degranulation, leukotriene C4 secretion, and calcium influx via mitochondrial calcium dysfunction. Free Radic. Biol. Med. 2010;49(4):632–640.Inoue T., Suzuki Y., Ra C. Epigallocatechin-3-gallate inhibits mast cell degranulation, leukotriene C4 secretion, and calcium influx via mitochondrial calcium dysfunction. Free Radic. Biol. Med. 2010;49(4):632–640.Rådinger M., Jensen B.M., Swindle E., Gilfillan A.M. Assay of mast cell mediators. Mast cells. Methods in molecular biology, vol 1220. Eds. M.Hughes and K. McNagny. N.Y.: Humana Press; 2015:307–323.Rådinger M., Jensen B.M., Swindle E., Gilfillan A.M. Assay of mast cell mediators. Mast cells. Methods in molecular biology, vol 1220. Eds. M.Hughes and K. McNagny. N.Y.: Humana Press; 2015:307–323.Falcone F.H., Wan D., Barwary N., SagiEisenberg R. RBL cells as models for in vitro studies of mast cells and basophils. Immunol. Rev. 2018;282(1):47–57.Falcone F.H., Wan D., Barwary N., SagiEisenberg R. RBL cells as models for in vitro studies of mast cells and basophils. Immunol. Rev. 2018;282(1):47–57.Turner H., Kinet J.P. Signalling through the high-affinity IgE receptor Fc epsilonRI. Nature. 1999;402(6760):B24–B30.Turner H., Kinet J.P. Signalling through the high-affinity IgE receptor Fc epsilonRI. Nature. 1999;402(6760):B24–B30.Saitoh S.I., Odom S., Gomez G., Sommers C.L., Young H.A., Rivera J., Samelson L.E. The four distal tyrosines are required for LAT-dependent signaling in FcepsilonRI-mediated mast cell activation. J. Exp. Med. 2003;198(5):831–843.Saitoh S.I., Odom S., Gomez G., Sommers C.L., Young H.A., Rivera J., Samelson L.E. The four distal tyrosines are required for LAT-dependent signaling in FcepsilonRI-mediated mast cell activation. J. Exp. Med. 2003;198(5):831–843.Fitzsimmons C.M., Falcone F.H., Dunne D.W. Helminth allergens, parasite-specific IgE, and its protective role in human immunity. Front. Immunol. 2014;5:61.Fitzsimmons C.M., Falcone F.H., Dunne D.W. Helminth allergens, parasite-specific IgE, and its protective role in human immunity. Front. Immunol. 2014;5:61.Marquardt D.L., Walker L.L. Dependence of mast cell IgE-mediated cytokine production on nuclear factor-kappaB activity. J. Allergy Clin. Immunol. 2000;105(3):500–505.Marquardt D.L., Walker L.L. Dependence of mast cell IgE-mediated cytokine production on nuclear factor-kappaB activity. J. Allergy Clin. Immunol. 2000;105(3):500–505.Murakami M., Taketomi Y. Secreted phospholipase A2 and mast cells. Allergol. Int. 2015;64(1):4–10.Murakami M., Taketomi Y. Secreted phospholipase A2 and mast cells. Allergol. Int. 2015;64(1):4–10.Gilfillan A.M., Rivera J. The tyrosine kinase network regulating mast cell activation. Immunol. Rev. 2009;228(1):149–169.Gilfillan A.M., Rivera J. The tyrosine kinase network regulating mast cell activation. Immunol. Rev. 2009;228(1):149–169.Tagen M., Elorza A., Kempuraj D., Boucher W., Kepley C.L., Shirihai O.S., Theoharides T.C. Mitochondrial uncoupling protein 2 inhibits mast cell activation and reduces histamine content. J. Immunol. 2009;183(10):6313–6319.Tagen M., Elorza A., Kempuraj D., Boucher W., Kepley C.L., Shirihai O.S., Theoharides T.C. Mitochondrial uncoupling protein 2 inhibits mast cell activation and reduces histamine content. J. Immunol. 2009;183(10):6313–6319.Li J., Miller E.J., Ninomiya-Tsuji J., Russell R.R., Young L.H. AMP-activated protein kinase activates p38 mitogen-activated protein kinase by increasing recruitment of p38 MAPK to TAB1 in the ischemic heart. Circ. Res. 2005;97(9):872–879.Li J., Miller E.J., Ninomiya-Tsuji J., Russell R.R., Young L.H. AMP-activated protein kinase activates p38 mitogen-activated protein kinase by increasing recruitment of p38 MAPK to TAB1 in the ischemic heart. Circ. Res. 2005;97(9):872–879.Lanna A., Henson S.M., Escors D., Akbar A.N. The kinase p38 activated by the metabolic regulator AMPK and scaffold TAB1 drives the senescence of human T cells. Nat. Immunol. 2014;15(10):965–972.Lanna A., Henson S.M., Escors D., Akbar A.N. The kinase p38 activated by the metabolic regulator AMPK and scaffold TAB1 drives the senescence of human T cells. Nat. Immunol. 2014;15(10):965–972.Demine S., Renard P., Arnould T. Mitochondrial uncoupling: a key controller of biological processes in physiology and diseases. Cells. 2019;8(8):795.Demine S., Renard P., Arnould T. Mitochondrial uncoupling: a key controller of biological processes in physiology and diseases. Cells. 2019;8(8):795.Tong S., Zhou T., Meng Y., Xu D., Chen J. AMPK decreases ERK1/2 activity and cancer cell sensitivity to nutrition deprivation by mediating a positive feedback loop involving eEF2K. Oncol. Lett. 2020;20(1):61–66.Tong S., Zhou T., Meng Y., Xu D., Chen J. AMPK decreases ERK1/2 activity and cancer cell sensitivity to nutrition deprivation by mediating a positive feedback loop involving eEF2K. Oncol. Lett. 2020;20(1):61–66.Meng R., Pei Z., Zhang A., Zhou Y., Cai X., Chen B., Liu G., Mai W., Wei J., Dong Y. AMPK activation enhances PPARα activity to inhibit cardiac hypertrophy via ERK1/2 MAPK signaling pathway. Arch. Biochem. Biophys. 2011;511(1–2):1–7.Meng R., Pei Z., Zhang A., Zhou Y., Cai X., Chen B., Liu G., Mai W., Wei J., Dong Y. AMPK activation enhances PPARα activity to inhibit cardiac hypertrophy via ERK1/2 MAPK signaling pathway. Arch. Biochem. Biophys. 2011;511(1–2):1–7.Hwang S.L., Li X., Lu Y., Jin Y., Jeong Y.T., Kim Y.D., Lee I.K., Taketomi Y., Sato H., Cho Y.S., Murakami M., Chang H.W. AMP-activated protein kinase negatively regulates FcεRI-mediated mast cell signaling and anaphylaxis in mice. J. Allergy Clin. Immunol. 2013;132(3):729–736.Hwang S.L., Li X., Lu Y., Jin Y., Jeong Y.T., Kim Y.D., Lee I.K., Taketomi Y., Sato H., Cho Y.S., Murakami M., Chang H.W. AMP-activated protein kinase negatively regulates FcεRI-mediated mast cell signaling and anaphylaxis in mice. J. Allergy Clin. Immunol. 2013;132(3):729–736.Lin K.C., Huang D.Y., Huang D.W., Tzeng S.J., Lin W.W. Inhibition of AMPK through Lyn-Syk-Akt enhances FcεRI signal pathways for allergic response. J. Mol. Med. 2016;94(2):183–194.Lin K.C., Huang D.Y., Huang D.W., Tzeng S.J., Lin W.W. Inhibition of AMPK through Lyn-Syk-Akt enhances FcεRI signal pathways for allergic response. J. Mol. Med. 2016;94(2):183–194.Chelombitko M.A., Fedorov A.V., Ilyinskaya O.P., Zinovkin R.A., Chernyak B.V. Role of reactive oxygen species in mast cell degranulation. Biochemistry (Mosc.). 2016;81(12):1564–1577.Chelombitko M.A., Fedorov A.V., Ilyinskaya O.P., Zinovkin R.A., Chernyak B.V. Role of reactive oxygen species in mast cell degranulation. Biochemistry (Mosc.). 2016;81(12):1564–1577.Jang J. Y., Min J.H., Chae Y.H., Baek J.Y., Wang S.B., Park S.J., Oh G.T., Lee S.H., Ho Y.S., Chang T.S. Reactive oxygen species play a critical role in collagen-induced platelet activation via SHP-2 oxidation. Antioxid. Redox Signal. 2014;20(16):2528–2540.Jang J. Y., Min J.H., Chae Y.H., Baek J.Y., Wang S.B., Park S.J., Oh G.T., Lee S.H., Ho Y.S., Chang T.S. Reactive oxygen species play a critical role in collagen-induced platelet activation via SHP-2 oxidation. Antioxid. Redox Signal. 2014;20(16):2528–2540.Yoo S.K., Starnes T.W., Deng Q., Huttenlocher A. Lyn is a redox sensor that mediates leukocyte wound attraction in vivo. Nature. 2011;480(7375):109–112.Yoo S.K., Starnes T.W., Deng Q., Huttenlocher A. Lyn is a redox sensor that mediates leukocyte wound attraction in vivo. Nature. 2011;480(7375):109–112.

The authors declare that there are no conflicts of interest present.