Endothelial protective properties of sacubitril-valsartan and drotaverine in influenza A(H1N1)pdm09 virus infection (Orthomyxoviridae: Alphainfluenzavirus: influenza A virus)

Cover Image

Cite item

Full Text

Abstract

Introduction. Severe influenza is characterized by damage and morphofunctional changes of the endothelium of blood vessels, which contributes to the development of hemorrhagic syndrome and can cause endothelial dysfunction. To optimize the pathogenetic therapy of influenza infection, a screening of medications with endothelial protective properties was carried out.

Aim. The aim of the study was to evaluate the endothelial protective properties of sacubitril/valsartan and drotaverine in influenza A(H1N1)pdm09 virus infection.

Materials and methods. The study was conducted on Wistar rats, which received sacubitril/valsartan and drotaverine (in treatment and prophylactic regimen) followed by intransal infection with influenza A/Saint-Petersburg/48/16 (H1N1)pdm09 virus. Rats without drug administration were included in the control group; rats received no medications followed by influenza virus infection – in challenge control group. The virus infectious activity was determined in pulmonary and mesenteric tissues. Vascular endothelium damage in lungs was assessed by three parameters (desquamation, morphological and dystrophic changes). Endothelial nitric oxide synthase (eNOS) expression level in endothelium of mesenteric blood vessels was determined as well as mesenteric arteries vasomotor activity.

Results. Drotaverine reduces the severity of histopathological changes in the pulmonary vascular endothelium; increases the maximal response of mesenteric blood vessels to acetylcholine compared to the infection control group; normalizes eNOS expression levels. Sacubitril/valsartan reduces the severity of desquamation in the pulmonary vascular endothelium; normalizes the response of mesenteric blood vessels to acetylcholine, while eNOS expression is decreased.

Conclusions. Drotaverine possesses more significant endothelial protective properties than sacubitril/valsartan when used in a treatment and prophylactic regimen in rats infected with influenza A(H1N1)pdm09 virus.

About the authors

Vladimir A. Marchenko

North-Western State Medical University Named after I.I. Mechnikov

Author for correspondence.
Email: vmarcenco@mail.ru
ORCID iD: 0000-0001-6870-3157

Ph. D. in medicine, Associate Professor of Medical Microbiology Department

Russian Federation, 191015, St. Petersburg

Irina A. Zelinskaya

V.A. Almazov National Medical Research Centre, Russian Ministry of Health

Email: irina.selinskaja@gmail.com
ORCID iD: 0000-0002-1971-3444

Researcher of Laboratory of Bioprosthetics and Cardiac Protection, Institute of Experimental Medicine

Russian Federation, 197341, St. Petersburg

Darya V. Mukhametdinova

V.A. Almazov National Medical Research Centre, Russian Ministry of Health

Email: mukh.dv@yandex.ru
ORCID iD: 0000-0002-7109-1187

Research Assistant of Experimental Pathomorphology Group, Institute of Experimental Medicine

Russian Federation, 197341, St. Petersburg

Yana G. Toropova

V.A. Almazov National Medical Research Centre, Russian Ministry of Health

Email: yana.toropova@mail.ru
ORCID iD: 0000-0003-1629-7868

Sc. D., Head of Laboratory of Bioprosthetics and Cardiac Protection, Institute of Experimental Medicine

Russian Federation, 197341, St. Petersburg

Michael M. Galagudza

V.A. Almazov National Medical Research Centre, Russian Ministry of Health

Email: galagoudza@mail.ru
ORCID iD: 0000-0001-5129-9944

MD, Sc. D., Professor and Corresponding member of the Russian Academy of Sciences, Director of the Institute of Experimental Medicine

Russian Federation, 197341, St. Petersburg

Irina N. Zhilinskaya

North-Western State Medical University Named after I.I. Mechnikov

Email: s_zhilinskaya@mail.ru
ORCID iD: 0000-0002-0084-1323

Sc. D., Professor of Medical Microbiology Department

Russian Federation, 191015, St. Petersburg

References

  1. WHO. Global Influenza Strategy 2019–2030; 2019. Available at: https://apo.org.au/node/224416
  2. Tenforde M.W., Noah K.P., O’Halloran A.C., Kirley P.D., Hoover C., Alden N.B., et al. Timing of influenza antiviral therapy and risk of death in adults hospitalized with influenza-associated pneumonia, influenza hospitalization surveillance network (FluSurv-NET), 2012–2019. Clin. Infect. Dis. 2025; 80(2): 461–8. https://doi.org/10.1093/cid/ciae427
  3. Lampejo T. Influenza and antiviral resistance: an overview. Eur. J. Clin. Microbiol. Infect. Dis. 2020; 39(7): 1201–8. doi: 10.1007/s10096-020-03840-9
  4. Fage C., Loison S., Zwygart A.C., Poli R., Rosset S., Medaglia C., et al. Influenza A(H1N1)pdm09 virus resistance to baloxavir, oseltamivir and sialic acid mimetics in single and dual therapies: Insights from human airway epithelia and murine models. Antiviral. Res. 2025; 239: 106174. https://doi.org/10.1016/j.antiviral.2025.106174
  5. Barber H. A case of influenzal myocarditis. Clin J. 1947; 76(5): 181–3.
  6. Skaarup K.G., Modin D., Nielsen L., Jensen J.U.S., Biering-Sørensen T. Influenza and cardiovascular disease pathophysiology: strings attached. Eur. Heart J. Suppl. 2023; 25(Suppl. A): A5–11. https://doi.org/10.1093/eurheartjsupp/suac117
  7. Jeyanathan T., Overgaard C., McGeer A. Cardiac complications of influenza infection in 3 adults. CMAJ. 2013; 185(7): 581–4. https://doi.org/10.1503/cmaj.110807
  8. Goldsteyn E.M. Influenza-associated mortality for circulatory and respiratory causes during the 2013-2014 through the 2018-2019 influenza seasons in Russia. Mezhdunarodnyi zhurnal prikladnykh i fundamental’nykh issledovanii. 2019; (12): 9–16. https://doi.org/10.17513/mjpfi.12945 https://elibrary.ru/dhthqt (in Russian)
  9. Cioffi D.L., Pandey S., Alvarez D.F., Cioffi E.A. Terminal sialic acids are an important determinant of pulmonary endothelial barrier integrity. Am. J. Physiol. Lung. Cell Mol. Physiol. 2012; 302(10): L1067–77. https://doi.org/10.1152/ajplung.00190.2011
  10. Vlasov T.D., Petrischev N.N., Lazovskaya O.A. Endothelial dysfunction. Do we understand this term properly? Vestnik anesteziologii i reanimatologii. 2020; 17(2): 76–84. https://doi.org/10.21292/2078-5658-2020-17-2-76-84 https://elibrary.ru/eqepoi (in Russian)
  11. Marchenko V.A., Zhilinskaya I.N. Endothelial activation and dysfunction caused by influenza A virus (Alphainfluenzavirus influenzae). Voprosy virusologii. 2024; 69(6): 465–78. https://doi.org/10.36233/0507-4088-264 https://elibrary.ru/zujoza (in Russian)
  12. Loscalzo J., Welch G. Nitric oxide and its role in the cardiovascular system. Prog. Cardiovasc. Dis. 1995; 38(2): 87–104. https://doi.org/10.1016/s0033-0620(05)80001-5
  13. Marchenko V.A., Barashkova S.V., Zelinskaya I.A., Toropova Ya.G., Ramsay E.S., Zhilinskaya I.N. Modulation of endothelial factors activity in human endothelial cells in influenza A(H1N1)pdm09 virus infection. Voprosy virusologii. 2021; 66(3): 198–210. https://doi.org/10.36233/0507-4088-48 https://elibrary.ru/wsxlvb (in Russian)
  14. Marchenko V.A., Zelinskaya I.A., Toropova Ya.G., Mukhametdinova D.V., Galagudza M.M., Lioznov D.A., et al. Duration of systemic alteration in vasomotor function of microvascular endothelium caused by the influenza A(H1N1)pdm09 virus. Regionarnoe krovoobrashchenie i mikrotsirkulyatsiya. 2023; 22(4): 74–86. https://doi.org/10.24884/1682-6655-2023-22-4-74-86 https://elibrary.ru/mmwnsf (in Russian)
  15. Roganova I.V. Comparative characteristics of cerebral circulation dysfunction in different age groups of patients with influenza virus infection. Izvestiya vysshikh uchebnykh zavedenii. Povolzhskii region. Meditsinskie nauki. 2011; (1): 108–15. https://elibrary.ru/oetuod (in Russian)
  16. Boytsov S.A. Influenza, novel Coronavirus infection and cardiovascular diseases. Kardiologicheskii vestnik. 2021; 16(1): 5–9. https://doi.org/10.17116/Cardiobulletin2021160115 https://elibrary.ru/zgvxkg (in Russian)
  17. Choudhary A., Rawat U., Kumar P., Mittal P. Pleotropic effects of statins: the dilemma of wider utilization of statin. Egypt. Heart J. 2023; 75(1): 1. https://doi.org/10.1186/s43044-023-00327-8
  18. Radigan K.A., Urich D., Misharin A.V., Chiarella S.E., Soberanes S., Gonzalez A., et al. The effect of rosuvastatin in a murine model of influenza A infection. PLoS One. 2012; 7(4): e35788. https://doi.org/10.1371/journal.pone.0035788
  19. Wu F., Wang C., Li S., Ye Y., Cui M., Liu Y., et al. Association between statins administration and influenza susceptibility: a systematic review and meta-analysis of longitudinal studies. Viruses. 2024; 16(2): 278. https://doi.org/10.3390/v16020278
  20. Caldeira D., Alarcão J., Vaz-Carneiro A., Costa J. Risk of pneumonia associated with use of angiotensin converting enzyme inhibitors and angiotensin receptor blockers: systematic review and meta-analysis. BMJ. 2012; 345: e4260. https://doi.org/10.1136/bmj.e4260
  21. Henry C., Zaizafoun M., Stock E., Ghamande S., Arroliga A.C., White H.D. Impact of angiotensin-converting enzyme inhibitors and statins on viral pneumonia. Proc. (Bayl. Univ. Med. Cent.). 2018; 31(4): 419–23. https://doi.org/10.1080/08998280.2018.1499293
  22. Sribhutorn A., Phrommintikul A., Wongcharoen W., Chaikledkaew U., Eakanunkul S., Sukonthasarn A. The modification effect of influenza vaccine on prognostic indicators for cardiovascular events after acute coronary syndrome: observations from an influenza vaccination trial. Cardiol. Res. Pract. 2016; 2016: 4097471. https://doi.org/10.1155/2016/4097471
  23. Marchenko V., Zelinskaya I., Toropova Y., Shmakova T., Podyacheva E., Lioznov D., et al. Influenza A virus causes histopathological changes and impairment in functional activity of blood vessels in different vascular beds. Viruses. 2022; 14(2): 396. https://doi.org/10.3390/v14020396
  24. Marchenko V., Mukhametdinova D., Amosova I., Lioznov D., Zhilinskaya I. Influenza A(H1N1)pdm09 virus alters expression of endothelial factors in pulmonary vascular endothelium in rats. Viruses. 2022; 14(11): 2518. https://doi.org/10.3390/v14112518
  25. Mazza A., Townsend D.M., Torin G., Schiavon L., Camerotto A., Rigatelli G., et al. The role of sacubitril/valsartan in the treatment of chronic heart failure with reduced ejection fraction in hypertensive patients with comorbidities: From clinical trials to real-world settings. Biomed. Pharmacother. 2020; 130: 110596. https://doi.org/10.1016/j.biopha.2020.110596
  26. Cassano V., Armentaro G., Magurno M., Aiello V., Borrello F., Miceli S., et al. Short-term effect of sacubitril/valsartan on endothelial dysfunction and arterial stiffness in patients with chronic heart failure. Front. Pharmacol. 2022; 13: 1069828. https://doi.org/10.3389/fphar.2022.1069828
  27. Hellmann M., Dąbrowska E., Żarczyńska-Buchowiecka M., Romanowska-Kocejko M., Narkiewicz K., Gruchała M., et al. Sacubitril/valsartan improved microvascular endothelial function in a young patient with COVID-19-related mild left ventricular dysfunction. Kardiol. Pol. 2022; 80(5): 614–5. https://doi.org/10.33963/kp.a2022.0063
  28. Ramakrishnan M.A. Determination of 50 % endpoint titer using a simple formula. World J. Virol. 2016; 5(2): 85–6. https://doi.org/10.5501/wjv.v5.i2.85
  29. Taylor C.R., Levenson R.M. Quantification of immunohistochemistry-issues concerning methods, utility and semiquantitative assessment II. Histopathology. 2006; 49(4): 411–24. https://doi.org/10.1111/j.1365-2559.2006.02513.x
  30. Marchenko V.A., Barashkova S.V., Zelinskaya I.A., Toropova Y.G., Sorokin E.V., Zhilinskaya I.N. Modeling influenza virus infection in mature Wistar rats. Voprosy virusologii. 2020; 65(3): 159–66. https://doi.org/10.36233/0507-4088-2020-65-3-159-166 https://elibrary.ru/ufdacp (in Russian)
  31. Janaszak-Jasiecka A., Siekierzycka A., Płoska A., Dobrucki I.T., Kalinowski L. Endothelial dysfunction driven by hypoxia – the influence of oxygen deficiency on no bioavailability. Biomolecules. 2021; 11(7): 982. https://doi.org/10.3390/biom11070982
  32. Zhang C. The role of inflammatory cytokines in endothelial dysfunction. Basic Res. Cardiol. 2008; 103(5): 398–406. https://doi.org/10.1007/s00395-008-0733-0
  33. Shaito A., Aramouni K., Assaf R., Parenti A., Orekhov A., Yazbi A.E., et al. Oxidative stress-induced endothelial dysfunction in cardiovascular diseases. Front. Biosci. (Landmark Ed). 2022; 27(3): 105. https://doi.org/10.31083/j.fbl2703105

Supplementary files

Supplementary Files
Action
1. JATS XML
Download
2. Fig. 1. Immunohistochemical assay of IAV NP expression in tissue and blood vessels of rat lungs and mesentery (magnification 200×). Pulmonary blood vessel of rat: a – group 1 (control); b – group 2 (IAV); c – group 3 (sacubitril/valsartan + IAV); d – group 4 (drotaverine + IAV); mesenteric blood vessel of rat: e – group 1 (control); f – group 2 (IAV); g – group 3 (sacubitril/valsartan + IAV); h – group 4 (drotaverine + IAV). Black arrows indicate NP-positive endothelial cells; white arrows denote NP-positive mast cells.

Download (1MB)
Indexing metadata
3. Fig. 2. Histological examination of pulmonary and mesenteric blood vessels (magnification 200 × for a–d, magnification 400 × for e–h, hematoxylin & eosin staining). Pulmonary blood vessel of rat: a – group 1 (control); b – group 2 (IAV); c – group 3 (sacubitril/valsartan + IAV); d – group 4 (drotaverine + IAV); mesenteric blood vessel of rat: e – group 1 (control); f – group 2 (IAV); g – group 3 (sacubitril/valsartan + IAV); h – group 4 (drotaverine + IAV).

Download (475KB)
Indexing metadata
4. Fig. 3. Histological alteration semi-quantitative score for pulmonary vascular endothelium in rats (M ± SE, score). * – p < 0.05 compared with group 1 (control); # – p < 0.05 compared with group 2 (IAV); & – p < 0.05 compared with group 3 (sacubitril/valsartan + IAV), Kruskal–Wallis test, 4–5 blood vessels from each animal (n = 5).

Download (252KB)
Indexing metadata
5. Fig. 4. eNOS expression level in mesenteric endothelium of rats, 3–5 blood vessels from each animal (n = 5). Mesenteric blood vessel of rat: a – group 1 (control); b – group 2 (IAV); c – group 3 (sacubitril/valsartan + IAV,); d – group 4 (drotaverine + IAV) (magnification 400×).

Download (134KB)
Indexing metadata
6. Fig. 5. eNOS expression level in mesenteric endothelium of rats (M ± SD, c.u.). * – p < 0.05 compared with group 1 (control), # – p < 0.05 compared with group 2 (IAV), & – p < 0.05 compared with group 3 (sacubitril/valsartan + IAV), Kruskal–Wallis test, 3–4 blood vessels from each animal (n = 5).

Download (236KB)
Indexing metadata
7. Fig. 6. Dose-dependent response curves of mesenteric arteries of rats (M ± SE). a – dose-dependent response to phenylephrine; b – dose-dependent response to acetylcholine. * – p < 0.05 compared with group 1 (control); # – p < 0.05 compared with group 2 (IAV), Brown-Forsythe test, 3 blood vessels from each animal (n = 5).

Download (388KB)
Indexing metadata
8. Fig. 7. Integral response of mesenteric arteries of rats (M ± SE). p > 0.05 compared with group 1 (control), Brown–Forsythe test, 3 blood vessels from each animal (n = 5).

Download (148KB)
Indexing metadata

Copyright (c) 2025 Marchenko V.A., Zelinskaya I.A., Mukhametdinova D.V., Toropova Y.G., Galagudza M.M., Zhilinskaya I.N.

Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.

Согласие на обработку персональных данных с помощью сервиса «Яндекс.Метрика»

1. Я (далее – «Пользователь» или «Субъект персональных данных»), осуществляя использование сайта https://journals.rcsi.science/ (далее – «Сайт»), подтверждая свою полную дееспособность даю согласие на обработку персональных данных с использованием средств автоматизации Оператору - федеральному государственному бюджетному учреждению «Российский центр научной информации» (РЦНИ), далее – «Оператор», расположенному по адресу: 119991, г. Москва, Ленинский просп., д.32А, со следующими условиями.

2. Категории обрабатываемых данных: файлы «cookies» (куки-файлы). Файлы «cookie» – это небольшой текстовый файл, который веб-сервер может хранить в браузере Пользователя. Данные файлы веб-сервер загружает на устройство Пользователя при посещении им Сайта. При каждом следующем посещении Пользователем Сайта «cookie» файлы отправляются на Сайт Оператора. Данные файлы позволяют Сайту распознавать устройство Пользователя. Содержимое такого файла может как относиться, так и не относиться к персональным данным, в зависимости от того, содержит ли такой файл персональные данные или содержит обезличенные технические данные.

3. Цель обработки персональных данных: анализ пользовательской активности с помощью сервиса «Яндекс.Метрика».

4. Категории субъектов персональных данных: все Пользователи Сайта, которые дали согласие на обработку файлов «cookie».

5. Способы обработки: сбор, запись, систематизация, накопление, хранение, уточнение (обновление, изменение), извлечение, использование, передача (доступ, предоставление), блокирование, удаление, уничтожение персональных данных.

6. Срок обработки и хранения: до получения от Субъекта персональных данных требования о прекращении обработки/отзыва согласия.

7. Способ отзыва: заявление об отзыве в письменном виде путём его направления на адрес электронной почты Оператора: info@rcsi.science или путем письменного обращения по юридическому адресу: 119991, г. Москва, Ленинский просп., д.32А

8. Субъект персональных данных вправе запретить своему оборудованию прием этих данных или ограничить прием этих данных. При отказе от получения таких данных или при ограничении приема данных некоторые функции Сайта могут работать некорректно. Субъект персональных данных обязуется сам настроить свое оборудование таким способом, чтобы оно обеспечивало адекватный его желаниям режим работы и уровень защиты данных файлов «cookie», Оператор не предоставляет технологических и правовых консультаций на темы подобного характера.

9. Порядок уничтожения персональных данных при достижении цели их обработки или при наступлении иных законных оснований определяется Оператором в соответствии с законодательством Российской Федерации.

10. Я согласен/согласна квалифицировать в качестве своей простой электронной подписи под настоящим Согласием и под Политикой обработки персональных данных выполнение мною следующего действия на сайте: https://journals.rcsi.science/ нажатие мною на интерфейсе с текстом: «Сайт использует сервис «Яндекс.Метрика» (который использует файлы «cookie») на элемент с текстом «Принять и продолжить».