antibioticsАнтибиотики и ХимиотерапияAntibiot Khimioter = Antibiotics and Chemotherapy0235-2990ООО «Издательство ОКИ»10.37489/0235-2990-2022-67-3-4-16-22antibiotics-908Research ArticleЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИExperimental ResearchВлияние рифампицина на индукцию активности MDR1/P-gp в провоспалительных макрофагах человекаThe Effect of Rifampicin on the Induction of MDR1/P-gp Activity in Proinflammatory Human Macrophageshttps://orcid.org/0000-0001-5662-3715ПавловаЕ. Н.PavlovaE. N.

 Павлова Екатерина Николаевна — аспирант биологического факультета

Москва

 Ekaterina N. Pavlova — post-graduate student of the Facultyof Biology

Moscow 

https://orcid.org/0000-0002-7256-4679ЕрохинаМ. В.ErokhinaM. V.

 Ерохина Мария Владиславовна — д. б. н., доцент, заместитель заведующего кафедрой клеточной биологии и гистологии

Каширское шоссе, 24,  г. Москва, 115522

 Maria V. Erokhina — D. Sc. in biology, Associate Professor

Moscow 

erokhina@bk.ruhttps://orcid.org/0000-0002-3068-0233РыбалкинаЕ. Ю.RybalkinaE. Yu.

 Рыбалкина Екатерина Юрьевна — старший научный сотрудник лаборатории генетики опухолевых клеток; старший научный сотрудник лаборатории клеточной биологии отдела патоморфологии, клеточной биологии и биохимии

Москва

 Ekaterina Yu. Rybalkina — Researcher at the Laboratory ofTumor Cell Genetics, Research Institute of Carcinogenesis; Senior Researcher at the Laboratory of Cell Biology, Department of Pathomorphology

Moscow 

https://orcid.org/0000-0002-0098-827XПоташниковаД. М.PotashnikovaD. M.

 Поташникова Дарья Марковна — к. б. н., старший научный сотрудник кафедры клеточной биологии и гистологии биологического факультета

Москва

 Daria M. Potashnikova — Ph. D. in biology

Moscow 

https://orcid.org/0000-0002-8067-4261МасютинА. Г.MasyutinA. G.

 Масютин Александр Георгиевич — научный сотрудник кафедры клеточной биологии и гистологии; научный сотрудник лаборатории клеточной биологии отдела патоморфологии, клеточной биологии и биохимии

 Москва

 

 Alexander G. Masyutin — Researcher at the Department ofCell Biology and Histology,  Researcher at the Laboratory of Cell Biology, Department of Pathomorphology, Cell Biology and Biochemistry

Moscow 

ЛепехаЛ. Н.LepekhaL. N.

 Лепеха Лариса Николаевна — д. б. н., профессор

Москва 

 Larisa N. Lepekha — D. Sc. in biology, Professor

Moscow 

https://orcid.org/0000-0002-2494-9275ЭргешовА. Э.ErgeshovA. E.

 Эргешов Атаджан Эргешович — д. м. н., профессор, директор

 Москва

 Atadzhan E. Ergeshov — D. Sc. in medicine, Professor

Moscow 

ФГБОУ ВО «Московский государственный университет им. М. В. Ломоносова»РоссияLomonosov Moscow State UniversityRussian FederationФГБОУ ВО «Московский государственный университет им. М. В. Ломоносова»; ФГБНУ «Центральный НИИ туберкулёза»РоссияLomonosov Moscow State University; Central Tuberculosis Research InstituteRussian FederationФГБНУ «Центральный НИИ туберкулёза»; ФГБУ «Национальный медицинский исследовательский центр онкологии им. Н. Н. Блохина Минздрава России»РоссияCentral Tuberculosis Research Institute; National Medical Research Center of Oncology named after N.N. Blokhin of the Ministry of Health of the Russian FederationRussian FederationФГБНУ «Центральный НИИ туберкулёза»РоссияCentral Tuberculosis Research InstituteRussian Federation202204082022673-41622Copyright © Павлова Е.Н., Ерохина М.В., Рыбалкина Е.Ю., Поташникова Д.М., Масютин А.Г., Лепеха Л.Н., Эргешов А.Э., 20222022Павлова Е.Н., Ерохина М.В., Рыбалкина Е.Ю., Поташникова Д.М., Масютин А.Г., Лепеха Л.Н., Эргешов А.Э.Pavlova E.N., Erokhina M.V., Rybalkina E.Y., Potashnikova D.M., Masyutin A.G., Lepekha L.N., Ergeshov A.E.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.antibiotics-chemotherapy.ru/jour/article/view/908

Актуальность. Влияние на активность белка множественной лекарственной устойчивости P-гликопротеина (P-gp, ген MDR1) в провоспалительных (М1) макрофагах человека рассматривается в качестве одной из перспективных стратегий повышения эффективности лечения больных туберкулёзом лёгких: активность P-gp является фактором, который может снижать внутриклеточное накопление рифампицина (RIF) — субстрата для P-gp.Цель работы: выявить влияние терапевтической концентрации RIF на активность P-gp в М1 макрофагах человека. Поставлены задачи: определить уровни экспрессии гена MDR1, белка P-gp и его функциональной активности на разных сроках дифференцировки клеток и при действии RIF.Материал и методы. В работе использованы следующие клеточные линии: суспензионные клетки промоноцитарной лейкемии ТНР-1 и индуцированные по провоспалительному фенотипу форболовым эфиром макрофаги ТНР-1. Суспензионные клетки миелобластной лейкемии К562/IS-9 с трансфицированным геном MDR1 использовались в качестве группы сравнения. Важным фактором является выбор экспериментальной концентрации RIF: у больных туберкулёзом лёгких средняя концентрация препарата составляет 10 мкг/мл. В работе использованы методы ОТ-ПЦР, иммуноцитохимии, проточной цитометрии.Результаты и обсуждение. Выявлена индукция экспрессии гена MDR1 в М1-макрофагах при краткосрочном воздействии «терапевтической» концентрации RIF. Этот эффект характерен только для макрофагов ТНР-1, в которых регистрируется значительная функциональная активность P-gp. В клетках, в которых активность P-gp не выявляется (суспензионные клетки ТНР-1), такая индукция не происходит. Это свидетельствует о наличие разных механизмов влияния RIF на MDR1, что может быть использовано для разработки стратегии ингибирования P-gp в макрофагах воспаления.Заключение. Учитывая ключевую роль макрофагов при туберкулёзе, необходима дальнейшая оценка MDR1/P-gp в операционном материале больных туберкулёзом лёгких, что позволит сделать вывод о необходимости разработки и применения лекарственных стратегий, направленный на блокировку функциональной активности P-gp и выборе более эффективных схем противотуберкулёзной терапии.

Background

Background. The effect on the activity of the multidrug resistance protein P-glycoprotein (P-gp, MDR1 gene) in pro-inflammatory (M1) human macrophages is considered one of the promising strategies for increasing the effectiveness of the treatment in patients with pulmonary tuberculosis: P-gp activity is considered a factor that reduces intracellular accumulation of rifampicin (RIF), a substrate for P-gp. The aim of this work was to reveal the effect of the therapeutic concentration of RIF on the activity of P-gp in M1 human macrophages. The objectives were as follows: to determine the expression levels of the MDR1 gene, P-gp protein, as well as its functional activity at different periods of cell differentiation and under the influence of RIF.

Material and methods

Material and methods. The following cell lines were used in the work: suspension cells of promonocytic leukemia THP-1 and THP-1 macrophages induced by phorbol ether according to the pro-inflammatory phenotype. Suspension cells of myeloid leukemia K562/IS-9 transfected with the MDR1 gene were used as a comparison group. An important factor is the choice of the experimental concentration of RIF: the average concentration of the drug in patients with pulmonary tuberculosis was 10 µg/ml. The methods of RT-PCR, immunocytochemistry, and flow cytometry were used in the work.

Results and discussion

Results and discussion. The induction of MDR1 gene expression in M1 macrophages under short-term exposure to a therapeutic concentration of RIF was revealed. This effect is typical only for THP-1 macrophages, in which a significant functional activity of P-gp is registered. This induction does not occur in the cells with no detectable P-gp activity (THP-1 suspension cells). This indicates the presence of different mechanisms of RIF influence on MDR1, which can be used to develop a strategy for P-gp inhibition in inflammatory macrophages.

Conclusion

Conclusion. Given the key role of macrophages in tuberculosis, further evaluation of MDR1/P-gp in the surgical material of patients with pulmonary tuberculosis is necessary, which makes it possible to draw a conclusion that it is necessary to develop and apply drug strategies aimed at blocking the functional activity of P-gp and choosing more effective anti-tuberculosis therapy regimens.

 

 

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