Cytotoxicity and anti-inflammatory properties of compounds isolated from Uromyrtus metrosideros
Description
The bioactivity of four compounds isolated from Uromyrtus metrosideros (numbered 3-6), and its polar crude extract (PCE) were tested in human peripheral blood mononuclear cells (PBMCs). First, cytotoxicity (or cell viability) of these compounds was tested at 10 μg/mL using the RealTime-Glo™ MT cell viability assay. Luminescence (RLU) was measured at 1, 6, and 24 h after treatment, using referencing the untreated samples as reference, and lysis buffer as a positive control. Next, human cytokine suppression by these four compounds and PCE was tested at 10 μg/mL. Inflammatory cytokines (IFN-γ, IL-17A, IL-8, and TNF) were induced in two ways. First, the PBMCs from three donors were activated with a cell stimulation cocktail of 50 ng/mL phorbol 12-myristate 13-acetate and 1 μg/mL ionomycin (P/I; eBioscience). Second, the PBMCs from four donors were cultured in the presence of Dynabeads Human T-Activator CD3/CD28 (Gibco). Cytokine concentrations were determined using a standard LEGENDplex™ human inflammation panel 1 (BioLegend®), LSRFortessa (BD), and LEGENDplex™ software (version 2022-02-10, Qognit, Inc.). PBMCs of three donors were used for Cell viability assay, and four donors for cytokine suppression assay. Compound details and variables in each dataset are provided in Compound details and Abbreviations.xlsx file. R scripts for statistical analyses and plots are provided for each experiment separately.