Perindopril Reduce SARS-COV-2 Spike-ACE2 Binding, ACE2 Overexpression and Cytokine Storm in Adipocyte Cell Infected with SARS-COV-2

Abstract

Abstract Funding Acknowledgements Type of funding sources: Public Institution(s). Main funding source(s): Ristekdikti Background and Aims The SARS-CoV-2 virus can infect adipocyte cells via ACE2 Receptor thus triggering ACE2 overexpression and cytokine storms which cause lethal complications. Hence, we explore the effect of Perindopril on the expression of ACE2, IL-6, IL-1B TNF-α in adipocyte cultures infected by SARS-CoV-2 spike protein and identify the possible mechanism involved. Materials and Methods Adipocyte culture obtained from a healthy and obese donor was divided into 4 triplicate groups (P0: negative control without treatment; P1: positive control (SARS-CoV-2 spike protein); P2: SARS-CoV-2 spike protein + exposure to Perindopril 0.5 µM); P3: SARS-CoV-2 spike protein + anti-ACE2 antibody 100 µg/mL; and evaluated at 24 and 48 hours. ACE2 expression was evaluated using immunofluorescence. IL-6, TNFα, and IL-1B were evaluated using ELISA. SARS-COV-2 Spike-ACE2 Binding was evaluated using Competitive ELISA. Data analysis was performed using SPSS 25.0 software. Results At first 24 hours of incubation, perindopril treatment has the highest ACE2 expression compared to negative control, positive control and anti-ACE2 antibody (113.52±0.34 ng/mL vs 13.3±0.87 ng/mL, 90.2±2.73 ng/mL, 17.3±0.11 ng/mL, p<0.01), lower ACE-ACE2R binding compared to anti-ACE2 antibody group (169.52±4.07 ng/mL vs 290.71±6.22 ng/mL, p<0.01) and higher IL-6 expression compared to positive control group (64.65±0.12 ng/mL vs 60.08±0.77 ng/mL p<0.01). Interestingly, after 48 hours, perindopril treatment was shown to prevent further increase of ACE2 expression compared to a positive control (47.37±0.76 ng/mL vs 80.31±5.37 ng/mL, p<0.01), higher SARS-COV-2 Spike-ACE2 binding compared to anti-ACE2 antibody group (143.68±3.68 ng/mL vs 103.1±9.49 ng/mL, p<0.01), and lower IL-6 expression compared to the positive control group (42.66±1.94 ng/mL vs 90.93±2.48 ng/mL p<0.01). However, no significant difference in TNFα and IL-1B expression between perindopril treatment and positive control in both 24 and 48 hours. Conclusion This study showed that perindopril reduces cytokine storm by preventing ACE-2 and IL-6 overexpression via an increasing number of SARS-COV-2 Spike-ACE2 competitive binding in adipocyte culture infected with SARS-COV-2 spike protein. A further clinical trial is needed to prove the benefit of perindopril in obese patients with COVID-19.