Garcinia kola and garcinoic acid suppress SARS‐CoV ‐2 spike glycoprotein S1 ‐induced hyper‐inflammation in human PBMCs through inhibition of NF‐κB activation

  1. Olumayokun A Olajide
  2. Victoria U Iwuanyanwu
  3. Izabela Lepiarz‐Raba
  4. Alaa A Al‐Hindawi
  5. Mutalib A Aderogba
  6. Hazel L Sharp
  7. Robert J Nash

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Abstract

Symptoms and complications associated with severe SARS‐CoV‐2 infection such as acute respiratory distress syndrome (ARDS) and organ damage have been linked to SARS‐CoV‐2 spike protein S1‐induced increased production of pro‐inflammatory cytokines by immune cells. In this study, the effects of an extract of Garcinia kola seeds and garcinoic acid were investigated in SARS‐CoV‐2 spike protein S1‐stimulated human PBMCs. Results of ELISA experiments revealed that Garcinia kola extract (6.25, 12.5, and 25 μg/ml) and garcinoic acid (1.25, 2.5, and 5 μM) significantly reduced SARS‐CoV‐2 spike protein S1‐induced secretion of TNFα, IL‐6, IL‐1β, and IL‐8 in PBMCs. In‐cell western assays showed that pre‐treatment with Garcinia kola extract and garcinoic acid reduced expressions of both phospho‐p65 and phospho‐IκBα proteins, as well as NF‐κB DNA binding capacity and NF‐κB‐driven luciferase expression following stimulation of PBMCs with spike protein S1. Furthermore, pre‐treatment of PBMCs with Garcinia kola extract prior to stimulation with SARS‐CoV‐2 spike protein S1 resulted in reduced damage to adjacent A549 lung epithelial cells. These results suggest that the seed of Garcinia kola and garcinoic acid are natural products which may possess pharmacological/therapeutic benefits in reducing cytokine storm in severe SARS‐CoV‐2 and other coronavirus infections.

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  1. Version published to 10.1002/ptr.7315
  2. ScreenIT

    SciScore for 10.1101/2021.05.18.444690: (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.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    The following antibodies were used: rabbit anti-phospho-p65 (Cell Signalling Technology), rabbit anti-IκBα (Cell Signalling Technology), and rabbit anti-phospho-IκBα (Santa Cruz Biotechnology) antibodies.
    rabbit anti-phospho-p65 (Cell Signalling Technology)
    suggested: None
    anti-phospho-p65
    suggested: None
    rabbit anti-IκBα (Cell Signalling Technology)
    suggested: None
    rabbit anti-phospho-IκBα (Santa Cruz Biotechnology)
    suggested: None
    anti-phospho-IκBα (Santa Cruz Biotechnology)
    suggested: None
    Thereafter, cells were washed with PBS and incubated with anti-rabbit HRP secondary antibody for 2 h at room temperature.
    anti-rabbit HRP
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Firefly and renilla luminescence intensities were measured using a FLUOstar OPTIMA microplate reader (BMG Labtech). 2.10 Co-culture of PBMCs with human A549 lung epithelial cells: Human A549 lung epithelial cells were co-cultured with PBMCs using the transwell system (0.4 μm porous membrane; Corning).
    A549
    suggested: None
    Software and Algorithms
    SentencesResources
    Statistical analysis were conducted using the GraphPad Prism software.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    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.
    • No funding statement was detected.
    • No protocol registration statement was detected.

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

    About SciScore

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  3. Version published to 10.1101/2021.05.18.444690v1 on bioRxiv