Hemin shows antiviral activity in vitro , possibly through suppression of viral entry mediators

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Heme oxygenase-1 (HO-1) is a stress-induced enzyme that catalyzes the breakdown of heme into biliverdin, carbon monoxide, and iron. Targeting HO-1 to treat severe COVID-19 has been suggested by several groups, yet the role of HO-1 in SARS-CoV-2 infection remains unclear. Based on this, we aimed to investigate the antiviral activity of Hemin, an activator of HO-1. Infectivity of SARS-CoV-2 was decreased in Vero E6 cells treated with Hemin. Hemin also decreased TMPRSS2 and ACE2 mRNA levels in non-infected cells, possibly explaining the observed decrease in infectivity. TMPRSS2 protein expression and proteolytic activity were decreased in Vero E6 cells treated with Hemin. Besides that, experimental studies supported with in silico calculations. Overall, our study supports further exploration of Hemin as a potential antiviral and inflammatory drug for the treatment of COVID-19.

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  1. SciScore for 10.1101/2022.05.24.493187: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    BlindingThe binding affinity energies (ΔG, kcal·mol-1) of Hemin with proteins were calculated with blind docking (supplementary material, DockingRawData.zip).
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Vero E6 cells were harvested as control, solvent control and drug treatment groups with Trypsin-EDTA (2X) and single cell suspensions were stained using an antibody against TMPRSS2 (Santa Cruz, sc-515727).
    suggested: (Santa Cruz Biotechnology Cat# sc-515727, RRID:AB_2892118)
    The monoclonal anti-ACE2 antibody (ProSci, Poway, CA,1:750 diluted in 5% BSA blocking buffer) and the goat anti-rabbit IgG-horseradish peroxidase (HRP)-conjugated antibody (Santa Cruz Biotechnology, Dallas, TX, 1:2500) were used as primary and secondary antibodies, respectively, for ACE2 protein expression.
    suggested: None
    anti-rabbit IgG-horseradish
    suggested: None
    The monoclonal anti-β-actin antibody (SantaCruz,1:1500) and the goat anti-mouse HRP conjugated (Pierce, Rockford, IL) were used as primary and secondary antibodies, respectively, for β-actin expression.
    suggested: None
    anti-mouse HRP
    suggested: None
    Experimental Models: Cell Lines
    Cell and Virus Culture: The cell culture media used for Vero E6 (African green monkey, kidney epithelial) cells consisted of high glucose Dulbecco’s modified Eagle medium (DMEM) (Capricorn) with 10 % fetal bovine serum (FBS) (Biolegend), 100 U/mL of penicillin (Lonza) and 0.1 mg/mL of streptomycin (Lonza).
    Vero E6
    suggested: None
    Software and Algorithms
    Results were evaluated statistically using the Two-way ANOVA (ordinary) technique (Prism 8 v8.2.1, GraphPad).
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Data analysis was performed using NovoExpress
    suggested: None

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    Results from scite Reference Check: We found no unreliable references.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.