Abstract

Coronavirus disease 2019 (COVID-19) is still a global public health emergency, although the efficacy of vaccines, no effective treatments are available. COVID-19 induces a cytokine storm, driving COVID-19 progression, severity, and mortality. Bronchial epithelial cells are the first pulmonary cells activated by coronavirus-2 (Sars-COV-2), leading to massive cytokine release, which hyperactivates lung fibroblasts, resulting in pulmonary fibrosis, a phenomenon observed even in moderate COVID-19 survivors. Therefore, the present in vitro study tested the hypothesis that Virlaza?, an herbal medicine, inhibits the hyperactivation of human bronchial epithelial cells (BEAS-2B) and pulmonary fibroblasts (MRC-5) induced by Sars-COV-2. BEAS-2B (5x104/mL/well) and MRC-5 (5x104/mL/well) cells were cocultured with 1 ml of blood from a Sars-COV-2-infected patient for 4 hours, and Virlaza? (1 µg/mL) was added in the first minute of the coculture. After 4 hours, the cells were recovered and used for analysis of cytotoxicity by a MTT assay and mRNA expression of the P2X7 receptor and iNOS. The supernatant was used to measure ATP and cytokine levels. Sars-COV-2 incubation resulted in increased release of ATP, IL-1beta, IL-6, IL-8, and TNF-alpha by BEAS-2B and MRC-5 cells (p<0.001). Treatment with Virlaza? resulted in a reduction in ATP, IL-1beta, IL-6, IL-8, and TNF-alpha release (p<0.001). In addition, Sars-COV-2 incubation resulted in increased expression of the P2X7 receptor and iNOS (p<0.001), which was reversed by Virlaza? (p<0.001). In conclusion, Virlaza? presents important anti-inflammatory effects in the context of Sars-COV-2 infection.