mimmunМедицинская иммунологияMedical Immunology (Russia)1563-06252313-741XSPb RAACI10.15789/1563-0625-NUT-3151mimmun-3151Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESНетрин-1 активирует экспрессию собственного гена и гена своего рецептора unc5b, но подавляет экспрессию гена ccl19 в макрофагах человека линии THP-1Netrin-1 upregulates the its own gene expression and unc5b gene expression, but downregulates ccl19 gene expression in human macrophages THP-1 cell lineНекрасоваЕ. В.NekrasovaE. V.

Некрасова Е.В. – научный сотрудник лаборатории регуляции липидного обмена 

197022, Санкт-Петербург, ул. Акад. Павлова, 12.

Nekrasova E.V., Researcher, Laboratory of Regulation of Lipid Metabolism 

12 Acad. Pavlov St St. Petersburg 197022

katherina.nekrasova@gmail.comОрловС. В.OrlovS. V.

Орлов С.В. – к.б.н., доцент, старший научный сотрудник лаборатории регуляции липидного обмена; доцент кафедры эмбриологии биологического факультета 

197022, Санкт-Петербург, ул. Акад. Павлова, 12.

Orlov S.V., PhD (Biology), Associate Professor, Senior Researcher, Laboratory of Regulation of Lipid Metabolism; Associate Professor, Department of Embryology Biology Faculty 

12 Acad. Pavlov St St. Petersburg 197022

ФГБНУ «Институт экспериментальной медицины»РоссияInstitute of Experimental MedicineRussian FederationФГБНУ «Институт экспериментальной медицины»; ФГБОУ ВО «Санкт-Петербургский государственный университет»РоссияInstitute of Experimental Medicine; St. Petersburg State UniversityRussian Federation20251510202527510131020Copyright © Некрасова Е.В., Орлов С.В., 20252025Некрасова Е.В., Орлов С.В.Nekrasova E.V., Orlov S.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.mimmun.ru/mimmun/article/view/3151

По данным Всемирной организации здравоохранения, за 2021 год основной причиной инвалидности и смертности людей во всем мире была ишемическая болезнь сердца, на третьем месте по числу смертей – инсульт. Данные сердечно-сосудистые заболевания, в том числе инфаркт миокарда и аневризмы стенки аорты, обусловлены системным хроническим воспалительным процессом под названием атеросклероз. Это заболевание развивается на протяжении десятилетий и характеризуется нарушением целостности эндотелиального слоя сосудов, избыточным накоплением липидов в стенках сосудов, дисфункцией макрофагов и гладкомышечных клеток и развитием воспаления артериальных стенок. Одним из путей регрессии атеросклероза может служить эмиграция макрофагов, нагруженных эфирами холестерола, из атеросклеротической бляшки по лимфатическим сосудам в региональные лимфоузлы и, в дальнейшем, в печень для последующего метаболизма холестерола. Белок нетрин-1 является ингибитором миграции иммунных клеток. Представляет интерес изучение аутокринной/паракринной регуляции экспрессии гена нетрина-1 в макрофагах человека. Понимание этого процесса может являться основой для поиска новых подходов в терапии атеросклероза. Цель исследования – изучить влияние белка нетрина-1 на уровень экспрессии генов ntn1, его рецептора unc5b и хемокина ccl19. Работа выполнена на макрофагах, дифференцированных из линии острой моноцитарной лейкемии человека THP-1. Анализ экспрессии генов ntn1, unc5b, ccl19 на уровне матричной РНК проведен методами обратной транскрипции и количественной полимеразной цепной реакции в режиме реального времени, а на уровне белка анализ сделан методом проточной цитофлуориметрии. Нетрин-1 в концентрации 50 нг/мл индуцировал экспрессию генов ntn1 и unc5b, но подавлял экспрессию гена ccl19 в макрофагах человека. Однако в концентрации 250 нг/мл нетрин-1 приводил к значительному снижению экспрессии собственного гена ntn1. Нетрин-1 оказывает ауторегулирующее влияние на экспрессию своего гена и гена своего рецептора unc5b в макрофагах человека. Подавление нетрином-1 экспрессии хемокина ccl19 в макрофагах может являться одним из путей ингибирования эмиграции макрофагов данным эффектором. Полученные результаты расширяют представление о регуляции экспрессии белков, ответственных за миграцию макрофагов, и в перспективе могут быть использованы для поиска новых мишеней в терапии атеросклероза.

According to the WHO data of 2021, ischemic heart disease was the leading cause of human disability and mortality at a global level, with brain stroke being the third leading cause of death. These cardiovascular diseases, including myocardial infarction and aortic aneurysms, are largely caused by atherosclerosis, a systemic chronic inflammatory process. This disease develops over decades and is characterized by the disruption of the vascular endothelial layer integrity, excessive lipid accumulation in arterial walls, macrophage and smooth muscle cells dysfunction, and evolving inflammation of arterial walls. One of approaches to atherosclerosis regression may be emigration of cholesterol ester-laden macrophage from atherosclerotic plaque via lymphatic vessels to the regional lymph nodes, and then to liver for subsequent cholesterol metabolism. The protein netrin-1 is inhibitor of immune cells migration. Particular interest is the study of the autocrine/paracrine regulation of netrin-1 gene expression in human macrophages. The understanding of this process may provide a basis for discovery of the new approaches to atherosclerosis therapy. The aim of the present article was to study the influence of protein factor netrin-1 on the gene expression levels of ntn1, its receptor unc5b, and ccl19. The experiments were carried out on the macrophage-like cells derived from acute monocytic leukemia cell line THP-1. The expression of ntn1, unc5b, ccl19 genes was performed by reverse transcription and real-time quantitative PCR (mRNA level), and by flow cytometry (at the protein level). Netrin-1 at the concentration of 50 ng/mL induced ntn1 and unc5b gene expression, but suppressed ccl19 gene expression in human macrophages. However, netrin-1 applied at the concentration of 250 ng/mL caused a significant downregulation of the own ntn1 gene expression. Netrin-1 has autoregulatory effect on its own gene expression, and on the gene encoding its receptor (unc5b) in human macrophage-like cells. The downregulation of chemokine ccl19 expression in macrophages mediated by netrin-1 may be a potential way by which this effector inhibits macrophage emigration. The obtained results suggest a regulatory mechanism for expression of proteins responsible for macrophage migration, which in the future will enable application of these data for searching new targets in atherosclerosis therapy.

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