Riboflavin and Thymine from Abutilon indicum Linn. Mitigate Allergic Responses: An Integrated network pharmacology, In-silico analysis, MD Simulation, In vitro and Ex vivo Studies

Abutilon indicum Linn., commonly known as Indian mallow, is known for its diverse pharmacological properties. This study investigates the anti-allergic and protective effects of hydro alcoholic extract of A. indicum (HAEAI) and two bioactive compounds, riboflavin and thymine, from A. indicum, using both in vitro and in vivo models. Cytotoxicity of these molecules was evaluated through the MTT assay on RBL-2H3 cells. Additionally, their anti-allergic effects were assessed using an ex vivo study, evaluating mast cell degranulation induced by Compound 48/80 in rat mesentery and the percentage protection of mast cells was quantified. Ionomycin was used to stimulate RBL-2H3 cells and mediator release. The degranulation levels of the compounds were assessed using the [Formula: see text]-hexosaminidase and histamine release assay. Additionally, the expression levels of cytokines, including TNF-[Formula: see text], IL-4, IL-6 and IL-13, were quantified using ELISA. The network consisting of composition, targets, and disease has been established. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed. Molecular docking techniques were used to simulate the binding affinity of active compounds to target proteins, while molecular dynamics were applied to validate the results obtained from the molecular docking. Riboflavin, Thymine and extract were found to be non-toxic on RBL-2H3 cells. Results demonstrated inhibition of the allergic response induced by Ionomycin. Hence, these two compounds present in Abutilon indicum Linn., emerged as potential drug candidates and can be used in the management of allergic conditions.