Calcio-Herbal Medicine Divya-Swasari-Vati Ameliorates SARS-CoV-2 Spike Protein-Induced Pathological Features and Inflammation in Humanized Zebrafish Model by Moderating IL-6 and TNF-α Cytokines
Authors: Acharya Balkrishna
Sudeep Verma
Siva Kumar Solleti
Anurag Varshney
Lakshmipathi Khandrika
Journal: Journal of Inflammation Research (2020)
Discipline: IMMUNOLOGY
Keywords: A549 xenotransplant; Divya-Swasari-Vati; SARS-CoV-2 infection model; ayurveda; behavioral fever
Affiliations: Drug Discovery and Development Division, Patanjali Research Institute, Haridwar, Uttarakhand 249 405, India., Department of Allied and Applied Sciences, University of Patanjali, Haridwar, Uttarakhand 249 405, India.

Abstract:  Purpose: Severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) infection has grown into a pandemic and without a specific cure, disease management is the need of the hour through symptomatic interventions. Studies with severe acute respiratory syndrome-coronavirus (SARS-CoV) have highlighted the role of herbal medicines either in combination with antiviral drugs or by themselves in curtailing the severity of infection and associated inflammation. Divya-Swasari-Vati is an Indian ayurvedic formulation used in the treatment of chronic cough and lung inflammation, which is one of the first symptoms of SARS-CoV-2 infections. Methods: In this study, we used a A549 cell xenotransplant in the swim bladder of zebrafish and modeled the SARS-CoV-2 infection by injecting the fish with a recombinant spike protein. The different groups were given normal feed or feed mixed with either dexamethasone (as the control drug) or Divya-Swasari-Vati. The changes in behavioral fever, infiltration of pro-inflammatory cells in the swim bladder, degeneration or presence of necrotic cells in the kidney, and gene expression of pro-inflammatory cytokines were studied to determine the rescue of the diseased phenotype. Results: Challenge with the spike protein caused changes in the swim bladder cytology with infiltrating pro-inflammatory cells, skin hemorrhage, and increase in behavioral fever. This was also accompanied by increased mortality of the disease control fish. Treatment with Divya-Swasari-Vati reversed most of the disease symptoms including damage to the kidney glomerulocytes, and complete reversal of behavioral fever. Dexamethasone, used as a comparator, was only able to partly rescue the behavioral fever phenotype. Divya-Swasari-Vati also suppressed the pro-inflammatory cytokines, IL-6 and TNF-α, levels in a dose-dependent manner, under in vivo and in vitro conditions. Conclusion: The study showed that the A549 xenotransplanted zebrafish injected with the recombinant spike protein of SARS-CoV-2 is an efficient model for the disease; and treatment with Divya-Swasari-Vati medicine rescued most of the inflammatory damage caused by the viral spike protein while increasing survival of the experimental fish. 

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 (more precisely, a  calcio-herbal medicine , as DSV contains calcium oxide, quicklime). Also by  behavioural fever , quantified by the fishes preference for a warmer environment. The validity of this approach is open to question. One question remains. In [a slightly earlier paper] , [Balkrishna et al (2020)] , the same authors applied the same experimental paradigm to show that a *different* herbal formulation (from the same manufacturer) *also* prevents a piscine cytokine storm and cures COVID-19. Are there any ayurveda-inspired products which **don t** cure COVD-19? We also read,  **Disclosure** The authors declare no conflicts of interest for this work. This is arrant nonsense. First author Acharya Balkrishna is co-owner of the company manufacturing DSV and co-owner of the company distributing it. The other authors are his employees. The publishers do themselves no credit by accepting his specious denial of any COI. ,