pulmoПульмонологияPULMONOLOGIYA0869-01892541-9617Scientific and Practical Journal “PULMONOLOGIYA” LLC10.18093/0869-0189-2017-27-2-187-197pulmo-842Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL STUDIESДиагностика заболеваний легких на основе протеомного анализа конденсата выдыхаемого воздухаDiagnosis of respiratory diseases using the proteomic analysis of exhaled breath condensateАнаевЭ. Х.AnaevE. Kh.el-anaev@hotmail.comФедорченкоК. Ю.FedorchenkoK. Yu.fedorchenko-kris@mail.ruКушаеваМ. Э.KushaevaM. E.dr-miya@mail.ruРябоконьА. М.Ryabokon'A. M.amryabokon@gmail.comКононихинА. С.KononikhinA. S.konoleha@yandex.ruБарминВ. В.BarminV. V.vitaly.barmin@gmail.comПикинО. В.PikinO. V.pikin-ov@mail.ruПоповИ. А.PopovI. A.hexapole@gmail.comНиколаевЕ. Н.NikolaevE. N.ennikolaev@rambler.ruВарфоломеевС. Д.VarfolomeevS. D.sdvarf@sky.chph.ras.ruЧучалинА. Г.ChuchalinA. G.pulmo-fmba@mail.ruФедеральное государственное бюджетное учреждение «Научно-исследовательский институт пульмонологии Федерального медико-биологического агентства России»РоссияFederal Pulmonology Research Institute, Federal Medical and Biological Agency of RussiaRussian FederationФедеральное государственное бюджетное образовательное учреждение высшего образования «Московский государственный университет имени М.В.Ломоносова»; Федеральное государственное бюджетное учреждение «Институт биохимической физики имени Н.М.Эмануэля» Российской академии наукРоссияM.V.Lomonosov Moscow Federal State University; N.M.Emanuel' Federal State Institute of Biochemical Physics, Russian Academy of SciencesRussian FederationФедеральное государственное бюджетное учреждение «Институт биохимической физики имени Н.М.Эмануэля» Российской академии наукРоссияN.M.Emanuel' Federal State Institute of Biochemical Physics, Russian Academy of SciencesRussian FederationФедеральное государственное бюджетное учреждение «Московский научно-исследовательский онкологический институт имени П.А.Герцена» Министерства здравоохранения Российской ФедерацииРоссияP.A.Gertsen Moscow Federal Research Oncology Institute, Healthcare Ministry of Russian FederationRussian Federation201705072017272187197Copyright © Анаев Э.Х., Федорченко К.Ю., Кушаева М.Э., Рябоконь А.М., Кононихин А.С., Бармин В.В., Пикин О.В., Попов И.А., Николаев Е.Н., Варфоломеев С.Д., Чучалин А.Г., 20172017Анаев Э.Х., Федорченко К.Ю., Кушаева М.Э., Рябоконь А.М., Кононихин А.С., Бармин В.В., Пикин О.В., Попов И.А., Николаев Е.Н., Варфоломеев С.Д., Чучалин А.Г.Anaev E.K., Fedorchenko K.Y., Kushaeva M.E., Ryabokon' A.M., Kononikhin A.S., Barmin V.V., Pikin O.V., Popov I.A., Nikolaev E.N., Varfolomeev S.D., Chuchalin A.G.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://journal.pulmonology.ru/pulm/article/view/842

Исследование конденсата выдыхаемого воздуха (КВВ) является неинвазивным методом диагностики заболеваний органов дыхания. Протеомный анализ КВВ – перспективный метод диагностики, позволяющий понять патологические механизмы и выявить различные фенотипы легочных заболеваний. Цель. Сравнительное изучение белкового спектра КВВ у здоровых добровольцев и пациентов с хронической обструктивной болезнью легких (ХОБЛ), пневмонией и немелкоклеточным раком легкого (НМРЛ), а также оценка возможности использования протеомного анализа КВВ для диагностики и дифференциальной диагностики этих заболеваний. Материалы и методы. Обследованы лица с ХОБЛ (n = 18), внебольничной пневмонией (n = 13), НМРЛ (n = 26) и здоровые некурящие добровольцы (n = 24). КВВ собран стандартизованным методом с помощью аппарата ECoScreen (Viasys Healthcare, Германия). Лиофилизированные и подвергнутые гидролизу трипсином образцы КВВ проанализированы с помощью нанопоточной высокоэффективной жидкостной хроматографии и тандемной масс-спектрометрии. Для поиска и идентификации белков использованы предоставленные Европейским институтом биоинформатики базы данных Mascot (Matrix Science, Великобритания) и IPI-human (version 3.82). Результаты. При протеомном анализе КВВ обследованных доноров 4 групп выявлено > 300 различных белков, бoльшую часть которых составляют цитоскелетные кератины I и II типов. Отмечено значительно более высокое содержание некоторых кератинов (5, 6 и 14) в образцах КВВ больных НМРЛ по сравнению с таковым у здоровых добровольцев. В КВВ у здоровых добровольцев, а также у больных ХОБЛ и пневмонией идентифицированы дермцидин, иммуноглобулин-α, кининоген, цитоплазматический актин, сывороточный альбумин, цинк-α2-гликопротеин. Высокое содержание пероксиредоксина в КВВ у больных ХОБЛ указывает на выраженный окислительный стресс. В образцах КВВ у пациентов с пневмонией обнаружен высокий уровень белков острой фазы воспаления и гипоксии (аннексины A1 и A2, HSP90B, цистатины М и В, фрагменты коллагенов и гистонов). В КВВ у больных НМРЛ определены β- и α-субъединицы гемоглобина, ядерный убиквитиновый казеин, POTEE, белки группы высокой мобильности (HMG-I / HMG-Y), лактоферрин. Заключение. В образцах КВВ у здоровых людей, больных ХОБЛ, пневмонией и НМРЛ обнаружен характерный белковый спектр. Большинство выявленных белков могут быть предложены в качестве панели для диагностики указанных заболеваний.

Investigation of exhaled breath condensate (EBC) is a noninvasive diagnostic method in respiratory diseases. The objective of this study was to compare EBC protein spectrum in healthy volunteers and in patients with chronic obstructive pulmonary disease (COPD), pneumonia and lung cancer (NSCLC), as well as to assess a role of proteomic analysis of EBC for diagnosis and differential diagnosis of these diseases. Methods. We examined 18 patients with COPD, 13 patients with community-acquired pneumonia, 26 patients with lung cancer and 24 healthy non-smoking volunteers. EBC was collected using ECoScreen system (VIASYS Healthcare, Germany) and a standardized method. EBC-samples were lyophilized, hydrolyzed and analyzed by HPLC and tandem mass spectrometry. To identify proteins, we used Mascot (Matrix Science, UK) and IPI-human (version 3.82) databases provided by the European Bioinformatics Institute. Results. Proteomic analysis of EBC identified more than 300 different proteins; most of them were types I and II cytoskeletal keratins. Cytokeratin 5, 6, and 14 concentrations in EBC of NSCLC patients were significantly higher than that in healthy volunteers. Dermcidin, immunoglobulin alpha, kininogen, cytoplasmic actin, serum albumin, and Zn-alpha2-glycoprotein were identified in EBC of healthy volunteers and patients with COPD and pneumonia. High concentration of peroxiredoxin in EBC of COPD patients could be due to severe oxidative stress. High levels of acute-phase and hypoxia proteins (annexins A1 and A2, HSP90B, cystatins M and B, collagen and histones fragments) were detected in EBC of pneumonia patients. Also, β- и α-subunit of hemoglobin, nuclear ubiquitin casein (NUCKS), POTEE, high mobility group protein (HMG-I/HMG-Y) and lactoferrin were identified in EBC of NSCLC patients. Conclusion. We found that EBC in healthy nonsmokers and in patients with COPD, pneumonia and NSCLC had characteristic protein spectrum. Most of the identified proteins could be used for diagnosis of these diseases.

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