sibmedСибирский научный медицинский журналСибирский научный медицинский журнал2410-25122410-2520ИЦиГ СО РАН10.18699/SSMJ20240614sibmed-1851Research ArticleМЕДИКО-БИОЛОГИЧЕСКИЕ НАУКИBIOMEDICINEРанозаживляющие свойства химерной формы GM-CSF человека на модели ожоговой травмы у крысWound healing properties of a chimeric form of human GM-CSF in a rat burn injury modelhttps://orcid.org/0000-0002-7808-9274ПыхтинаМ. Б.PykhtinaM. B.

Пыхтина Мария Борисовна - к.б.н.

630060, Новосибирск, ул. Тимакова, 2

Maria B. Pykhtina - сandidate of biological sciences.

630117, Novosibirsk, Timakova st., 2

pykhtina_maria@mail.ruhttps://orcid.org/0000-0002-6740-8241МирошниченкоС. М.MiroshnichenkoS. M.

Мирошниченко Светлана Михайловна.

630060, Новосибирск, ул. Тимакова, 2

Svetlana M. Miroshnichenko.

630117, Novosibirsk, Timakova st., 2

svmiro@yandex.ruhttps://orcid.org/0000-0002-8200-3998КарповМ. А.KarpovM. A.

Карпов Михаил Александрович - к.м.н.

630060, Новосибирск, ул. Тимакова, 2; 630091, Новосибирск, Красный пр., 52

Mikhail A. Karpov - сandidate of medical sciences.

630117, Novosibirsk, Timakova st., 2; 630091, Novosibirsk, Krasny ave., 52

karpov-ma@mail.ruhttps://orcid.org/0000-0002-8036-7990АбышевА. A.AbyshevA. A.

Абышев Александр Андреевич.

630060, Новосибирск, ул. Тимакова, 2; 630091, Новосибирск, Красный пр., 52

Alexandr A. Abyshev.

630117, Novosibirsk, Timakova st., 2; 630091, Novosibirsk, Krasny ave., 52

abyshev.alexander@mail.ruhttps://orcid.org/0000-0001-8968-3968МосалевК. И.MosalevK. I.

Мосалев Кирилл Игоревич.

630060, Новосибирск, ул. Тимакова, 2

Kirill I. Mosalev.

630117, Novosibirsk, Timakova st., 2

mosalevkir@mail.ruhttps://orcid.org/0000-0001-6276-9630КотоваМ. В.KotovaM. V.

Котова Мария Владимировна.

630060, Новосибирск, ул. Тимакова, 2

Maria V. Kotova.

630117, Novosibirsk, Timakova st., 2

zerokiri@mail.ruhttps://orcid.org/0000-0001-9732-1246БеклемишевА. Б.BeklemishevA. A.

Беклемишев Анатолий Борисович - д.б.н.

630060, Новосибирск, ул. Тимакова, 2

Anatoly B. Beklemishev - doctor of biological sciences.

630117, Novosibirsk, Timakova st., 2

ab.beklem-46@yandex.ruhttps://orcid.org/0000-0003-2678-8783КнязевР. А.KnyazevR. A.

Князев Роман Александрович - к.б.н.

630060, Новосибирск, ул. Тимакова, 2

Roman A. Knyazev - сandidate of biological sciences.

630117, Novosibirsk, Timakova st., 2

knjazev_roman@mail.ruФИЦ фундаментальной и трансляционной медициныРоссияFederal Research Center of Fundamental and Translational MedicineRussian FederationФИЦ фундаментальной и трансляционной медицины; Новосибирский государственный медицинский университет Минздрава РоссииРоссияFederal Research Center of Fundamental and Translational Medicine; Novosibirsk State Medical University of Minzdrav of RussiaRussian Federation202408012025446146154Copyright © Пыхтина М.Б., Мирошниченко С.М., Карпов М.А., Абышев А.A., Мосалев К.И., Котова М.В., Беклемишев А.Б., Князев Р.А., 20252025Пыхтина М.Б., Мирошниченко С.М., Карпов М.А., Абышев А.A., Мосалев К.И., Котова М.В., Беклемишев А.Б., Князев Р.А.Pykhtina M.B., Miroshnichenko S.M., Karpov M.A., Abyshev A.A., Mosalev K.I., Kotova M.V., Beklemishev A.A., Knyazev R.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://sibmed.elpub.ru/jour/article/view/1851

Термические ожоги являются одной из распространенных видов травм человека, вызывая в тяжелых случаях обширный иммунный ответ как локально в ране, так и системно. Актуальной задачей остается поиск новых средств местного лечения ожоговых ран. Цель исследования – изучить ранозаживляющие свойства рекомбинантного гранулоцитарно-макрофагального колониестимулирующего фактора (ryGM-CSF), а также его пролонгированной химерной формы с аполипопротеином АI человека (ryGM-CSF-apoAI) на модели ожоговой травмы у крыс. Материал и методы. В работе использовали ryGM-CSF и ryGM-CSF-apoAI, полученные экспрессией в дрожжах P. рastoris. Исследование выполнялось на 27 крысах Wistar, которым моделировали ожоги III степени. Ранозаживляющее действие рекомбинантных цитокинов оценивали морфологически, а также определяли экспрессию генов цитокинов – маркеров течения раневого процесса и ангиогенеза. Результаты и их обсуждение. ryGM-CSF на модели ожога крыс проявлял как свойства цитокина, влияющего на ангиогенез, повышая количество сосудов, так и, являясь чужеродным белком, стимулировал иммунный ответ, увеличивая пролонгированно количество лимфоцитов, и поддерживал воспаление. Химерная форма цитокина ryGM-CSF-apoAI угнетала воспаление, ускоряла смену фаз ранозаживления и, как следствие, снижала количество нейтрофилов к 16-м суткам. Неожиданно она способствовала созреванию сосудов, которые в данном случае имели нормальный просвет, размер и были выстланы эндотелием. Соотношение содержания мРНК генов Angpt1 и Angpt2 было в 1,6 раза больше в случае применения ryGM-CSF-apoAI по сравнению с контролем и назначением ryGM-CSF. Заключение. При ксеногенном введении ryGM-CSF пролонгирует воспаление за счет раннего лимфоцитарного ответа на чужеродный белок и повышенного содержания нейтрофилов на более поздних сроках ранозаживления. Химерная форма ryGM-CSF-apoAI снижает иммуногенность цитокина, способствуя более ранней замене нейтрофилов на макрофаги и фибробласты и стимулирует созревание полноценных кровеносных сосудов.

Thermal burns are one of the common types of human injuries, causing in severe cases an extensive immune response both locally in the wound and having a systemic effect. The search for new means of local treatment of burn wounds remains an urgent task. The aim of this study was to investigate the wound healing properties of recombinant granulocyte-macrophage colony-stimulating factor (ryGM-CSF) and its prolonged chimeric form with human apolipoprotein AI (ryGM-CSF-apoAI) on the model of burn injury in rats. Material and methods. ryGM-CSF and ryGM-CSF-apoAI obtained by expression in yeast P. pastoris were used in this work. The study was performed on 27 Wistar rats, which were modeled burns of III degree. The wound-healing effect of recombinant cytokines was evaluated morphologically, and the expression of cytokine genes - markers of the course of the wound process and angiogenesis - was measured. Results and discussion. ryGM-CSF is known to accelerate wound healing. Human rGM-CSF on the rat burn model showed both the properties of a cytokine influencing angiogenesis, increasing the number of vessels, and being a foreign protein stimulated the immune response, increasing prolonged the number of lymphocytes and supporting inflammation. The chimeric form of cytokine ryGM-CSF-apoAI reduced inflammation, accelerated the change of wound healing phases and, as a consequence, reduced the number of neutrophils by 16 days. Unexpectedly, it induced maturation of vessels, which in this case had normal lumen, size, and were lined with endothelium. The ratio Angpt1 and Angpt2 gene mRNA content was 2-fold higher when ryGM-CSF-apoAI was used, compared to control and ryGM-CSF administration. Conclusions. With xenogeneous administration, recombinant human GM-CSF prolongs inflammation due to early lymphocytic response to foreign protein and increased neutrophil content at later wound healing periods. The chimeric form ryGM-CSF-apoAI reduces cytokine immunogenicity by promoting earlier replacement of neutrophils by macrophages and fibroblasts and stimulates maturation of high-grade blood vessels.

ожоговая ранакрысы Wistarрекомбинантный химерный GM-CSF человекааполипопротеин АI человекаэкспрессия генов цитокиноввоспалениеангиогенезburn woundWistar ratsrecombinant human chimeric GM-CSFhuman apolipoprotein AIcytokine gene expressioninflammationangiogenesisРабота выполнена в рамках темы НИР №1022072600029-5-1.6.2;1.6.8 с использованием оборудования ЦКП «Протеомный анализ», поддержанного финансированием Минобрнауки России (соглашение №075-15-2021-691).The work was carried out according to state assignment № 1022072600029-5-1.6.2;1.6.8 using the equipment of the Proteomic Analysis Collective Use Center, supported by funding from the Ministry of Education and Science of the Russian Federation (agreement No. 075-15-2021-691).ReferencesWHO. The global burden of disease 2004. Available at: https://iris.who.int/bitstream/handle/10665/43942/9789241563710_eng.pdfWHO. The global burden of disease 2004. 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The authors declare that there are no conflicts of interest present.