Effects of digoxin in inhibiting ACE2 and SARS-CoV-2 binding for attenuating COVID-19 in human adipocytes
Abstract
BACKGROUND Angiotensin-converting enzyme 2 (ACE2) has a role in SARS-CoV-2 incidence, and digoxin is a competitive inhibitor of SARS-CoV-2-ACE2 binding. This study aimed to investigate the effects of digoxin on SARS-CoV-2-ACE2 binding, proinflammatory cytokines, and prothrombotic factors in adipocytes of patients with COVID-19.
METHODS This in vitro study used adipocyte cultures, which were divided into negative control, positive control (SARS-CoV-2 S1 spike protein only), SARS-CoV-2 S1 spike protein with digoxin, and SARS-CoV-2 S1 spike protein with human recombinant soluble ACE2 (hrsACE2). Data were analyzed using one-way ANOVA and Pearson correlation.
RESULTS SARS-CoV-2 significantly elevated ACE2 and increased interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), tissue factor (TF), and plasminogen activator inhibitor-1 (PAI-1) compared to the negative control group (p<0.001). No SARS-CoV-2-ACE2 binding was detected in SARS-CoV-2 with digoxin and hrsACE2 groups, compared to the positive control group (0 ng/ml versus 0 ng/ml versus 36.33 [1.58] ng/ml, p<0.001). Digoxin significantly decreased IL-6 (48.94 [1.80] ng/ml versus 90.93 [4.29] ng/ml; p<0.001), TNF-α (87.65 [6.88] ng/ml versus 307.95 [57.34] ng/ml; p<0.001), TF (5.33 [0.32] ng/ml versus 6.85 [0.22] ng/ml; p<0.001), and PAI-1 levels (2.92 [0.168] ng/ml versus 4.86 [0.11] ng/ml; p<0.001), compared to positive control group. ACE2 positively correlated with IL-6 (p = 0.004, r = 0.763) and TF (p = 0.004, r = 0.768) but was not correlated with IL-1β, TNF-α, and PAI-1 levels.
CONCLUSIONS This study promoted digoxin therapy to prevent cytokine storm and thromboembolism by decreasing IL-6, TNF-α, TF, and PAI-1 in adipocyte cultured models at an early stage of COVID-19.
Downloads
References
Morens DM, Fauci AS. Emerging pandemic diseases: how we got to COVID-19. Cell. 2020;183(3):837. Erratum for: Cell. 2020;182(5):1077−92. https://doi.org/10.1016/j.cell.2020.08.021
Ren X, Zhou J, Guo J, Hao C, Zheng M, Zhang R, et al. Reinfection in patients with COVID-19: a systematic review. Glob Health Res Policy. 2022;7(1):12. https://doi.org/10.1186/s41256-022-00245-3
Ryan PM, Caplice NM. Is adipose tissue a reservoir for viral spread, immune activation, and cytokine amplification in coronavirus disease 2019? Obesity (Silver Spring). 2020;28(7):1191−4. https://doi.org/10.1002/oby.22843
Caohuy H, Eidelman O, Chen T, Liu S, Yang Q, Bera A, et al. Common cardiac medications potently inhibit ACE2 binding to the SARS-CoV-2 spike, and block virus penetration and infectivity in human lung cells. Sci Rep. 2021;11(22195). https://doi.org/10.1038/s41598-021-01690-9
Han T, Kang J, Li G, Ge J, Gu J. Analysis of 2019-nCoV receptor ACE2 expression in different tissues and its significance study. Ann Transl Med. 2020;8(17):1077. https://doi.org/10.21037/atm-20-4281
Yalcin HC, Sukumaran V, Al-Ruweidi MK, Shurbaji S. Do changes in ACE-2 expression affect SARS-CoV-2 virulence and related complications: a closer look into membrane-bound and soluble forms. Int J Mol Sci. 2021;22(13):6703. https://doi.org/10.3390/ijms22136703
El-Sayed Moustafa JS, Jackson AU, Brotman SM, Guan L, Villicaña S, Roberts AL, et al. ACE2 expression in adipose tissue is associated with cardio-metabolic risk factors and cell type composition-implications for COVID-19. Int J Obes (Lond). 2022;46(8):1478−86. https://doi.org/10.1038/s41366-022-01136-w
Whyte CS, Simpson M, Morrow GB, Wallace CA, Mentzer AJ, Knight JC, et al. The suboptimal fibrinolytic response in COVID-19 is dictated by high PAI-1. J Thromb Haemost. 2022;20(10):2394−406. https://doi.org/10.1111/jth.15806
Chen X, Zhao B, Qu Y, Chen Y, Xiong J, Feng Y, et al. Detectable serum severe acute respiratory syndrome coronavirus 2 viral load (RNAemia) is closely correlated with drastically elevated interleukin 6 level in critically ill patients with coronavirus disease 2019. Clin Infect Dis. 2020;71(8):1937−42. https://doi.org/10.1093/cid/ciaa449
Cho J, Lee YJ, Kim JH, Kim SI, Kim SS, Choi BS, et al. Antiviral activity of digoxin and ouabain against SARS-CoV-2 infection and its implication for COVID-19. Sci Rep. 2020;10(1):16200. https://doi.org/10.1038/s41598-020-72879-7
Carswell KA, Lee MJ, Fried SK. Culture of isolated human adipocytes and isolated adipose tissue. Methods Mol Biol. 2012;806:203−14. https://doi.org/10.1007/978-1-61779-367-7_14
Dosch SF, Mahajan SD, Collins AR. SARS coronavirus spike protein-induced innate immune response occurs via activation of the NF-kappaB pathway in human monocyte macrophages in vitro. Virus Res. 2009;142(1−2):19−27. https://doi.org/10.1016/j.virusres.2009.01.005
Gutiérrez-Chamorro L, Riveira-Muñoz E, Barrios C, Palau V, Nevot M, Pedreño-López S, et al. SARS-CoV-2 infection modulates ACE2 function and subsequent inflammatory responses in swabs and plasma of COVID-19 patients. Viruses. 2021;13(9):1715. https://doi.org/10.3390/v13091715
Patel SK, Juno JA, Lee WS, Wragg KM, Hogarth PM, Kent SJ, et al. Plasma ACE2 activity is persistently elevated following SARS-CoV-2 infection: implications for COVID-19 pathogenesis and consequences. Eur Respir J. 2021;57(5):2003730. https://doi.org/10.1183/13993003.03730-2020
Reindl-Schwaighofer R, Hödlmoser S, Eskandary F, Poglitsch M, Bonderman D, Strassl R, et al. ACE2 elevation in severe COVID-19. Am J Respir Crit Care Med. 2021;203(9):1191−6. https://doi.org/10.1164/rccm.202101-0142LE
Gubernatorova EO, Gorshkova EA, Polinova AI, Drutskaya MS. IL-6: relevance for immunopathology of SARS-CoV-2. Cytokine Growth Factor Rev. 2020;53:13−24. https://doi.org/10.1016/j.cytogfr.2020.05.009
Han H, Yang L, Liu R, Liu F, Wu KL, Li J, et al. Prominent changes in blood coagulation of patients with SARS-CoV-2 infection. Clin Chem Lab Med. 2020;58(7):1116−20. https://doi.org/10.1515/cclm-2020-0188
D'Elia JA, Bayliss G, Gleason RE, Weinrauch LA. Cardiovascular-renal complications and the possible role of plasminogen activator inhibitor: a review. Clin Kidney J. 2016;9(5):705−12. https://doi.org/10.1093/ckj/sfw080
Han M, Pandey D. ZMPSTE24 regulates SARS-CoV-2 spike protein-enhanced expression of endothelial PAI-1. Am J Respir Cell Mol Biol. 2021;65(3):300−8. https://doi.org/10.1165/rcmb.2020-0544OC
Klok FA, Kruip MJ, van der Meer NJ, Arbous MS, Gommers DA, Kant KM, et al. Incidence of thrombotic complications in critically ill ICU patients with COVID-19. Thromb Res. 2020;191:145−7. https://doi.org/10.1016/j.thromres.2020.04.013
Cui S, Chen S, Li X, Liu S, Wang F. Prevalence of venous thromboembolism in patients with severe novel coronavirus pneumonia. J Thromb Haemost. 2020;18(6):1421−4. https://doi.org/10.1111/jth.14830
Copyright (c) 2024 Meity Ardiana, I Gde Rurus Suryawan, Hanestya Oky Hermawan, Primasitha Maharani Harsoyo Putri, Safira Rahma
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Authors who publish with Medical Journal of Indonesia agree to the following terms:
- Authors retain copyright and grant Medical Journal of Indonesia right of first publication with the work simultaneously licensed under a Creative Commons Attribution-NonCommercial License that allows others to remix, adapt, build upon the work non-commercially with an acknowledgment of the work’s authorship and initial publication in Medical Journal of Indonesia.
- Authors are permitted to copy and redistribute the journal's published version of the work non-commercially (e.g., post it to an institutional repository or publish it in a book), with an acknowledgment of its initial publication in Medical Journal of Indonesia.
