cfpdБюллетень физиологии и патологии дыханияBulletin Physiology and Pathology of Respiration1998-5029Дальневосточный научный центр физиологии и патологии дыхания10.36604/1998-5029-2023-87-52-61cfpd-1074Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL RESEARCHПрофиль экспрессии муцинов MUC5AC и MUC5B у больных бронхиальной астмой на фоне холодового воздействияProfile of MUC5AC and MUC5B mucins expression in asthma patients under cold exposureНаумовД. Е.NaumovD. E.

Денис Евгеньевич Наумов - кандидат медицинских наук, зав. лабораторией, лаборатория молекулярных и трансляционных исследований.

675000, Благовещенск, ул. Калинина, 22

Denis E. Naumov - PhD (Med.), Head of Laboratory of Molecular and Translational Research.

22 Kalinina Str., Blagoveshchensk, 675000

denn1985@bk.ruФедеральное государственное бюджетное научное учреждение "Дальневосточный научный центр физиологии и патологии дыхания"Far Eastern Scientific Center of Physiology and Pathology of Respiration2023030420230875261Copyright © Наумов Д.Е., 20232023Наумов Д.Е.Naumov D.E.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://cfpd.elpub.ru/jour/article/view/1074Введение

Введение. Холодовая гиперреактивность дыхательных путей (ХГДП) – распространенное состояние у больных бронхиальной астмой (БА), ухудшающее течение заболевания и качество жизни пациентов. MUC5AC и MUC5B – основные секретируемые муцины в респираторном тракте, принимающие участие в обеспечении нормального мукоцилиарного клиренса, но также способные провоцировать развитие патологических изменений в случае дизрегуляции оптимального баланса их продукции.

Цель

Цель. Установить особенности экспрессии муцинов MUC5AC и MUC5B в динамике на фоне экспериментального охлаждения у больных БА в зависимости от статуса ХГДП. Материалы и методы. В исследование было включено 98 человек, в том числе 26 больных хроническим необструктивным бронхитом вне обострения (контрольная группа) и 72 больных БА. Экспрессию MUC5AC, MUC5B и TRPM8 определяли в верхних дыхательных путях методом количественной ПЦР с обратной транскрипцией. Продукцию MUC5AC и MUC5B также определяли в мокроте методом ИФА. Всем больным проводили бронхопровокационную пробу с изокапнической гипервентиляцией холодным воздухом для выявления ХГДП, а также аналогичную назальную пробу для оценки эффекта охлаждения на экспрессию изучаемых генов.

Результаты

Результаты. Больные БА отличались увеличенной в 4,22 раза экспрессией MUC5AC (p=0,02) в назальном эпителии по сравнению с лицами контрольной группы. ХГДП была ассоциирована с исходной апрегуляцией MUC5AC в 7,33 раза (p=0,008), а также с дальнейшим нарастанием экспрессии MUC5AC, но снижением MUC5B в ответ на охлаждение, чего не отмечалось у больных БА без ХГДП. Базальная экспрессия TRPM8 была взаимосвязана с исходным уровнем MUC5AC (ρ=0,41, p=0,04), MUC5B (ρ=0,55, p<0,001) и количеством отделяемой мокроты после холодовой бронхопровокационной пробы.

Заключение

Заключение. Больные БА с ХГДП демонстрируют более выраженный дисбаланс в продукции муцинов, усугубляющийся на фоне холодового воздействия. Это, в свою очередь, может приводить к ряду патологических нарушений, ассоциированных с более тяжелым течением заболевания.

Introduction

Introduction. Cold airway hyperresponsiveness (CAH) is a common condition in patients with asthma, which worsens the clinical course of the disease and the patients’ quality of life. MUC5AC and MUC5B are the main secreted mucins in the respiratory tract, which are involved in normal mucociliary clearance, but also capable of provoking the development of pathological changes in case of dysregulation of their balanced production.

Aim

Aim. The aim of this study was to determine the dynamics of MUC5AC and MUC5B expression during experimental cooling in patients with asthma depending on the status of CAH.

Materials and methods

Materials and methods. The study enrolled 98 subjects including 26 patients with chronic non-obstructive bronchitis without exacerbation (control group) and 72 patients with asthma. The expression of MUC5AC, MUC5B and TRPM8 was determined in the upper respiratory tract by quantitative reverse transcription PCR. The production of MUC5AC and MUC5B was also measured in sputum by ELISA. All patients underwent a bronchoprovocation test with isocapnic cold air hyperventilation to detect CAH, and a similar nasal challenge was performed to assess the effect of cooling on the expression of the studied genes.

Results

Results. Patients with asthma had 4.22-fold increase in the expression of MUC5AC (p=0.02) in the nasal epithelium as compared with the control group. CAH was associated with an initial 7.33-fold upregulation of MUC5AC (p=0.008) as well as with further increase in MUC5AC expression but a decrease in MUC5B in response to cooling, which was not observed in asthma patients without CAH. Basal TRPM8 expression was associated with baseline level of MUC5AC (ρ=0.41, p=0.04), MUC5B (ρ=0.55, p<0.001) and amount of sputum produced after the cold bronchoprovocation.

Conclusion

Conclusion. Asthma patients with CAH demonstrate a more pronounced imbalance in the production of mucins, which is aggravated by cold exposure. This, in turn, can lead to a number of pathological disorders associated with a more severe course of the disease.

бронхиальная астмахолодовая гиперреактивность дыхательных путейохлаждениеэкспрессиясекрециямуциныMUC5ACMUC5BTRPM8asthmacold-induced airway hyperresponsivenesscoolingexpression secretionmucinsMUC5ACMUC5BTRPM8Исследование выполнено при поддержке РФФИ (проект №17-54-53162)This study was supported by RFBR (project №17-5453162)ReferencesBonser L.R., Erle D.J. Airway Mucus and Asthma: The Role of MUC5AC and MUC5B // J. Clin. Med. 2017. Vol.6, Iss.12. Article number: 112. https://doi.org/10.3390/jcm6120112Bonser L.R., Erle D.J. Airway Mucus and Asthma: The Role of MUC5AC and MUC5B. J. Clin. Med. 2017; 6(12):112. https://doi.org/10.3390/jcm6120112Lillehoj E.P., Kato K., Lu W., Kim K.C. Cellular and molecular biology of airway mucins // Int. Rev. Cell Mol. Biol. 2013. Vol.303. P.139–202. https://doi.org/10.1016/B978-0-12-407697-6.00004-0Lillehoj E.P., Kato K., Lu W., Kim K.C. Cellular and molecular biology of airway mucins. Int. Rev. Cell Mol. Biol. 2013; 303:139–202. https://doi.org/10.1016/B978-0-12-407697-6.00004-0Song D., Cahn D., Duncan G.A. Mucin Biopolymers and Their Barrier Function at Airway Surfaces // Langmuir. 2020. Vol.36, Iss.43. P.12773–12783. https://doi.org/10.1021/acs.langmuir.0c02410Song D., Cahn D., Duncan G.A. Mucin Biopolymers and Their Barrier Function at Airway Surfaces. Langmuir 2020; 36(43):12773–12783. https://doi.org/10.1021/acs.langmuir.0c02410Martínez-Rivera C., Crespo A., Pinedo-Sierra C., García-Rivero J.L., Pallarés-Sanmartín A., Marina-Malanda N., Pascual-Erquicia S., Padilla A., Mayoralas-Alises S., Plaza V., López-Viña A., Picado C. Mucus hypersecretion in asthma is associated with rhinosinusitis, polyps and exacerbations // Respir. Med. 2018. Vol.135. P.22–28. https://doi.org/10.1016/j.rmed.2017.12.013Martínez-Rivera C., Crespo A., Pinedo-Sierra C., García-Rivero J.L., Pallarés-Sanmartín A., Marina-Malanda N., Pascual-Erquicia S., Padilla A., Mayoralas-Alises S., Plaza V., López-Viña A., Picado C. Mucus hypersecretion in asthma is associated with rhinosinusitis, polyps and exacerbations. Respir. Med. 2018; 135:22–28. https://doi.org/10.1016/j.rmed.2017.12.013Radicioni G., Ceppe A., Ford A.A., Alexis N.E., Barr R.G., Bleecker E.R., Christenson S.A., Cooper C.B., Han M.K., Hansel N.N., Hastie A.T., Hoffman E.A., Kanner R.E., Martinez F.J., Ozkan E., Paine R. 3rd, Woodruff P.G., O'Neal W.K., Boucher R.C., Kesimer M. Airway mucin MUC5AC and MUC5B concentrations and the initiation and progression of chronic obstructive pulmonary disease: an analysis of the SPIROMICS cohort // Lancet Respir. Med. 2021. Vol.9, Iss.11. P.1241–1254. https://doi.org/10.1016/S2213-2600(21)00079-5Radicioni G., Ceppe A., Ford A.A., Alexis N.E., Barr R.G., Bleecker E.R., Christenson S.A., Cooper C.B., Han M.K., Hansel N.N., Hastie A.T., Hoffman E.A., Kanner R.E., Martinez F.J., Ozkan E., Paine R. 3rd, Woodruff P.G., O'Neal W.K., Boucher R.C., Kesimer M. Airway mucin MUC5AC and MUC5B concentrations and the initiation and progression of chronic obstructive pulmonary disease: an analysis of the SPIROMICS cohort. Lancet Respir. Med. 2021; 9(11):1241– 1254. https://doi.org/10.1016/S2213-2600(21)00079-5Hyrkäs-Palmu H., Ikäheimo T.M., Laatikainen T., Jousilahti P., Jaakkola M.S., Jaakkola J.J.K. Cold weather increases respiratory symptoms and functional disability especially among patients with asthma and allergic rhinitis // Sci. Rep. 2018. Vol.8, Iss.1. Article number: 10131. https://doi.org/10.1038/s41598-018-28466-yHyrkäs-Palmu H., Ikäheimo T.M., Laatikainen T., Jousilahti P., Jaakkola M.S., Jaakkola J.J.K. Cold weather increases respiratory symptoms and functional disability especially among patients with asthma and allergic rhinitis. Sci. Rep. 2018; 8(1):10131 https://doi.org/10.1038/s41598-018-28466-yHan A., Deng S., Yu J., Zhang Y., Jalaludin B., Huang C. Asthma triggered by extreme temperatures: From epidemiological evidence to biological plausibility // Environ. Res. 2023. Vol.216, Pt2. Article number: 114489. https://doi.org/10.1016/j.envres.2022.114489Han A., Deng S., Yu J., Zhang Y., Jalaludin B., Huang C. Asthma triggered by extreme temperatures: From epidemiological evidence to biological plausibility. Environ. Res. 2023; 216(Pt2):114489. https://doi.org/10.1016/j.envres.2022.114489Zhu Y., Yang T., Huang S., Li H., Lei J., Xue X., Gao Y., Jiang Y., Liu C., Kan H., Chen R. Cold temperature and sudden temperature drop as novel risk factors of asthma exacerbation: a longitudinal study in 18 Chinese cities // Sci. Total Environ. 2022. Vol.814. Article number: 151959. https://doi.org/10.1016/j.scitotenv.2021.151959Zhu Y., Yang T., Huang S., Li H., Lei J., Xue X., Gao Y., Jiang Y., Liu C., Kan H., Chen R. Cold temperature and sudden temperature drop as novel risk factors of asthma exacerbation: a longitudinal study in 18 Chinese cities. Sci. Total Environ. 2022; 814:151959. https://doi.org/10.1016/j.scitotenv.2021.151959Liu S.C., Lu H.H., Fan H.C., Wang H.W., Chen H.K., Lee F.P., Yu C.J., Chu Y.H. The identification of the TRPM8 channel on primary culture of human nasal epithelial cells and its response to cooling // Medicine (Baltimore). 2017. Vol.96, Iss.31. Article number: e7640. https://doi.org/10.1097/MD.0000000000007640Liu S.C., Lu H.H., Fan H.C., Wang H.W., Chen H.K., Lee F.P., Yu C.J., Chu Y.H. The identification of the TRPM8 channel on primary culture of human nasal epithelial cells and its response to cooling. Medicine (Baltimore) 2017; 96(31):e7640. https://doi.org/10.1097/MD.0000000000007640Lachowicz-Scroggins M.E., Yuan S., Kerr S.C., Dunican E.M., Yu M., Carrington S.D., Fahy J.V. Abnormalities in MUC5AC and MUC5B Protein in Airway Mucus in Asthma // Am. J. Respir. Crit. Care Med. 2016. Vol.194, Iss.10. P.1296–1299. https://doi.org/10.1164/rccm.201603-0526LELachowicz-Scroggins M.E., Yuan S., Kerr S.C., Dunican E.M., Yu M., Carrington S.D., Fahy J.V. Abnormalities in MUC5AC and MUC5B Protein in Airway Mucus in Asthma. Am. J. Respir. Crit. Care Med. 2016; 194(10):1296–1299. https://doi.org/10.1164/rccm.201603-0526LEKiwamoto T., Katoh T., Evans C.M., Janssen W.J., Brummet M.E., Hudson S.A., Zhu Z., Tiemeyer M., Bochner B.S. Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis // J. Allergy Clin. Immunol. 2015. Vol.135, Iss.5. P.1329‒1340.e9. https://doi.org/10.1016/j.jaci.2014.10.027Kiwamoto T., Katoh T., Evans C.M., Janssen W.J., Brummet M.E., Hudson S.A., Zhu Z., Tiemeyer M., Bochner B.S. Endogenous airway mucins carry glycans that bind Siglec-F and induce eosinophil apoptosis. J. Allergy Clin. Immunol. 2015; 135(5):1329–1340.e9. https://doi.org/10.1016/j.jaci.2014.10.027Welsh K.G., Rousseau K., Fisher G., Bonser L.R., Bradding P., Brightling C.E., Thornton D.J., Gaillard E.A. MUC5AC and a Glycosylated Variant of MUC5B Alter Mucin Composition in Children With Acute Asthma // Chest. 2017. Vol.152, Iss.4. P.771–779. https://doi.org/10.1016/j.chest.2017.07.001Welsh K.G., Rousseau K., Fisher G., Bonser L.R., Bradding P., Brightling C.E., Thornton D.J., Gaillard E.A. MUC5AC and a Glycosylated Variant of MUC5B Alter Mucin Composition in Children With Acute Asthma. Chest 2017; 152(4):771–779. https://doi.org/10.1016/j.chest.2017.07.001Tajiri T., Matsumoto H., Jinnai M., Kanemitsu Y., Nagasaki T., Iwata T., Inoue H., Nakaji H., Oguma T., Ito I., Niimi A. Pathophysiological relevance of sputum MUC5AC and MUC5B levels in patients with mild asthma // Allergol. Int. 2022. Vol.71, Iss.2. P.193–199. https://doi.org/10.1016/j.alit.2021.09.003Tajiri T., Matsumoto H., Jinnai M., Kanemitsu Y., Nagasaki T., Iwata T., Inoue H., Nakaji H., Oguma T., Ito I., Niimi A. Pathophysiological relevance of sputum MUC5AC and MUC5B levels in patients with mild asthma. Allergol. Int. 2022; 71(2):193–199. https://doi.org/10.1016/j.alit.2021.09.003Evans C.M., Raclawska D.S., Ttofali F., Liptzin D.R., Fletcher A.A., Harper D.N., McGing M.A., McElwee M.M., Williams O.W., Sanchez E., Roy M.G., Kindrachuk K.N., Wynn T.A., Eltzschig H.K., Blackburn M.R., Tuvim M.J., Janssen W.J., Schwartz D.A., Dickey B.F. The polymeric mucin Muc5ac is required for allergic airway hyperreactivity // Nat. Commun. 2015. Vol.6. Article number: 6281. https://doi.org/10.1038/ncomms7281Evans C.M., Raclawska D.S., Ttofali F., Liptzin D.R., Fletcher A.A., Harper D.N., McGing M.A., McElwee M.M., Williams O.W., Sanchez E., Roy M.G., Kindrachuk K.N., Wynn T.A., Eltzschig H.K., Blackburn M.R., Tuvim M.J., Janssen W.J., Schwartz D.A., Dickey B.F. The polymeric mucin Muc5ac is required for allergic airway hyperreactivity. Nat. Commun. 2015; 6:6281. https://doi.org/10.1038/ncomms7281Некрасов Э.В., Перельман Ю.М., Приходько А.Г., Захарова Э.В., Макарова Г.А. Секреция муцинов слизистой носа в ответ на гипервентиляцию холодным воздухом у больных бронхиальной астмой с разной степенью тяжести и контролируемости заболевания // Бюллетень физиологии и патологии дыхания. 2014. Вып.53. С.42–49. EDN: SNECKH.Nekrasov E.V., Perelman J.M., Prikhodko А.G., Zakharova E.V., Makarova G.A. [Mucin secretion in the nasal mucosa in response to cold air hyperventilation in asthmatics with different degrees of asthma control and disease severity]. Bûlleten' fiziologii i patologii dyhaniâ = Bulletin Physiology and Pathology of Respiration 2014; (53):42‒49 (in Russian).Song D., Iverson E., Kaler L., Boboltz A., Scull M.A., Duncan G.A. MUC5B mobilizes and MUC5AC spatially aligns mucociliary transport on human airway epithelium // Sci. Adv. 2022. Vol.8, Iss.47. Article number: eabq5049. https://doi.org/10.1126/sciadv.abq5049Song D., Iverson E., Kaler L., Boboltz A., Scull M.A., Duncan G.A. MUC5B mobilizes and MUC5AC spatially aligns mucociliary transport on human airway epithelium. Sci. Adv. 2022; 8(47):eabq5049. doi: 10.1126/sciadv.abq5049Roy M.G., Livraghi-Butrico A., Fletcher A.A., McElwee M.M., Evans S.E., Boerner R.M., Alexander S.N., Bellinghausen L.K., Song A.S., Petrova Y.M., Tuvim M.J., Adachi R., Romo I., Bordt A.S., Bowden M.G., Sisson J.H., Woodruff P.G., Thornton D.J., Rousseau K., De la Garza M.M., Moghaddam S.J., Karmouty-Quintana H., Blackburn M.R., Drouin S.M., Davis C.W., Terrell K.A., Grubb B.R., O'Neal W.K., Flores S.C., Cota-Gomez A., Lozupone C.A., Donnelly J.M., Watson A.M., Hennessy C.E., Keith R.C., Yang I.V., Barthel L., Henson P.M., Janssen W.J., Schwartz D.A., Boucher R.C., Dickey B.F., Evans C.M. Muc5b is required for airway defence // Nature. 2014. Vol.505, Iss.7483. P.412–416. https://doi.org/10.1038/nature12807Roy M.G., Livraghi-Butrico A., Fletcher A.A., McElwee M.M., Evans S.E., Boerner R.M., Alexander S.N., Bellinghausen L.K., Song A.S., Petrova Y.M., Tuvim M.J., Adachi R., Romo I., Bordt A.S., Bowden M.G., Sisson J.H., Woodruff P.G., Thornton D.J., Rousseau K., De la Garza M.M., Moghaddam S.J., Karmouty-Quintana H., Blackburn M.R., Drouin S.M., Davis C.W., Terrell K.A., Grubb B.R., O'Neal W.K., Flores S.C., Cota-Gomez A., Lozupone C.A., Donnelly J.M., Watson A.M., Hennessy C.E., Keith R.C., Yang I.V., Barthel L., Henson P.M., Janssen W.J., Schwartz D.A., Boucher R.C., Dickey B.F., Evans C.M. Muc5b is required for airway defence. Nature 2014; 505(7483):412–416. https://doi.org/10.1038/nature12807Cho H.Y., Park S., Miller L., Lee H.C., Langenbach R., Kleeberger S.R. Role for Mucin-5AC in Upper and Lower Airway Pathogenesis in Mice // Toxicol. Pathol. 2021. Vol.49, Iss.5. P.1077–1099. https://doi.org/10.1177/01926233211004433Cho H.Y., Park S., Miller L., Lee H.C., Langenbach R., Kleeberger S.R. Role for Mucin-5AC in Upper and Lower Airway Pathogenesis in Mice. Toxicol Pathol. 2021; 49(5):1077–1099. https://doi.org/10.1177/01926233211004433Ehre C., Worthington E.N., Liesman R.M., Grubb B.R., Barbier D., O'Neal W.K., Sallenave J.M., Pickles R.J., Boucher R.C. Overexpressing mouse model demonstrates the protective role of Muc5ac in the lungs // Proc. Natl Acad. Sci. USA. 2012. Vol.109, Iss.41. P.16528–16533. https://doi.org/10.1073/pnas.1206552109Ehre C., Worthington E.N., Liesman R.M., Grubb B.R., Barbier D., O'Neal W.K., Sallenave J.M., Pickles R.J., Boucher R.C. Overexpressing mouse model demonstrates the protective role of Muc5ac in the lungs. Proc. Natl Acad. Sci. USA 2012; 109(41):16528–16533. https://doi.org/10.1073/pnas.1206552109Kirkham S., Sheehan J.K., Knight D., Richardson P.S., Thornton D.J. Heterogeneity of airways mucus: variations in the amounts and glycoforms of the major oligomeric mucins MUC5AC and MUC5B // Biochem. J. 2002. Vol.36, Pt3. P.537–546. https://doi.org/10.1042/0264-6021:3610537Kirkham S., Sheehan J.K., Knight D., Richardson P.S., Thornton D.J. Heterogeneity of airways mucus: variations in the amounts and glycoforms of the major oligomeric mucins MUC5AC and MUC5B. Biochem J. 2002; 361(Pt3):537– 546. https://doi.org/10.1042/0264-6021:3610537Наумов Д.Е., Котова О.О., Гассан Д.А., Афанасьева Е.Ю. Шелудько Е.Г. Взаимосвязь экспрессии гена катионных каналов TRPM8 с холодовой гиперреактивностью дыхательных путей у больных бронхиальной астмой // Бюллетень физиологии и патологии дыхания. 2019. Вып.72. С.33–38. EDN: GAVIVO. https://doi.org/10.12737/article_5d09d6a0d75552.76525437Naumov D.E., Kotova O.O., Gassan D.A., Afanas’eva E.Yu., Sheludko E.G. [Correlation of cation channel TRPM8 gene expression with cold-induced airway hyperresponsiveness in asthma patients]. Bûlleten' fiziologii i patologii dyhaniâ = Bulletin Physiology and Pathology of Respiration 2019; (72):33–38 (in Russian). https://doi.org/10.12737/article_5d09d6a0d75552.76525437

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