Application of Humanized Zebrafish Model in the Suppression of SARS-CoV-2 Spike Protein Induced Pathology by Tri-Herbal Medicine Coronil via Cytokine Modulation
Authors: Sudeep Verma
Acharya Balkrishna
Anurag Varshney
Siva Kumar Solleti
Journal: Molecules (2020)
Discipline: CHEMISTRY
Keywords: Ayurveda; Coronil; SARS-CoV-2 infection; anti-inflammatory; behavioral fever
Affiliations: Drug Discovery and Development Division, Patanjali Research Institute, NH-58, Haridwar 249 405, Uttarakhand, India., Department of Allied and Applied Sciences, University of Patanjali, Patanjali Yog Peeth, Roorkee-Haridwar Road, Haridwar 249 405, Uttarakhand, India.

Abstract:  Zebrafish has been a reliable model system for studying human viral pathologies. SARS-CoV-2 viral infection has become a global chaos, affecting millions of people. There is an urgent need to contain the pandemic and develop reliable therapies. We report the use of a humanized zebrafish model, xeno-transplanted with human lung epithelial cells, A549, for studying the protective effects of a tri-herbal medicine Coronil. At human relevant doses of 12 and 58 µg/kg, Coronil inhibited SARS-CoV-2 spike protein, induced humanized zebrafish mortality, and rescued from behavioral fever. Morphological and cellular abnormalities along with granulocyte and macrophage accumulation in the swim bladder were restored to normal. Skin hemorrhage, renal cell degeneration, and necrosis were also significantly attenuated by Coronil treatment. Ultra-high-performance liquid chromatography (UHPLC) analysis identified ursolic acid, betulinic acid, withanone, withaferine A, withanoside IV-V, cordifolioside A, magnoflorine, rosmarinic acid, and palmatine as phyto-metabolites present in Coronil. In A549 cells, Coronil attenuated the IL-1β induced IL-6 and TNF-α cytokine secretions, and decreased TNF-α induced NF-κB/AP-1 transcriptional activity. Taken together, we show the disease modifying immunomodulatory properties of Coronil, at human equivalent doses, in rescuing the pathological features induced by the SARS-CoV-2 spike protein, suggesting its potential use in SARS-CoV-2 infectivity. 

Retracted: No

PubPeer comments:
 The rationale of this study is modeled on lung-cancer research, modified along the following lines: - persuade human-sourced adenocarcinomic epithelial cells (A549 cell-line) to take up residence in zebrafish. - administer SARS-CoV-2 spike proteins. - Spike proteins will attack the A549 cells in the same way as the SARS-CoV-2 virus attacks normal epithelial respiratory cells in human hosts. - Zebrafish immune system will respond to that attack with a cytokine storm, in the same way as the human immune system responds to a SARS-CoV-2 infection. - An ad-hoc formulation of herbs will prevent that cytokine storm, in zebrafish and in humans, as quantified by the piscine swimbladder and kidney function. The validity of the study could be questioned, but there is no doubting its value for the manufacture of the herbal preparation (Coronil), who lost no time in exploiting this publication for marketing purposes. ------------------------------------ *[The company continues to insist its product works against Covid-19.] * * It has treated and cured people, Acharya Balkrishna, managing director of Patanjali told the BBC.* *It referred us to scientific trials, the results of which it says have been published in several peer-reviewed journals.* *It pointed specifically to a study from November 2020 in a journal published by the Swiss-based MDPI, which was based on a laboratory trial.* ------------------------------------------------------------------ One question remains. In [a slightly later paper] , [Balkrishna et al (2020)] , the same authors applied the same experimental paradigm to show that a *different*, pre-existing herbal formulation (from the same manufacturer) *also* prevents a piscine cytokine storm and cures COVID-19. If this is true, what is the rationale for creating a **new** herbal formulation? We also read,  **Conflicts of Interest** The authors declare no conflict of interest in the publication of the data in this manuscript. This is arrant nonsense. As reported by the BBC, first author Acharya Balkrishna -- co-owner of the company manufacturing Coronil, co-owner of the company distributing it -- is directly benefiting from this paper, using it to claim that his medication prevents deaths from COVID19. The co-authors are his employees. The publishers do themselves no credit by accepting his specious denial of any COI. ,