Cytotoxicity and anti-inflammatory properties of compounds isolated from Uromyrtus metrosideros

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.从铜色乌墨（Uromyrtus metrosideros）中分离得到的4种化合物（编号3~6）及其极性粗提物（polar crude extract, PCE）的生物活性，在人外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）中进行了检测。 首先，采用RealTime-Glo™ MT细胞活力检测法，以10 μg/mL的浓度测试了这些化合物的细胞毒性（或细胞活力）。以未处理样品作为参照，以裂解液作为阳性对照，分别于处理后1、6、24 h检测发光强度（Relative Light Unit, RLU）。 随后，以10 μg/mL的浓度测试了这4种化合物及PCE对人细胞因子的抑制活性。研究通过两种方式诱导炎性细胞因子（IFN-γ、IL-17A、IL-8及TNF）的产生：其一，将3名供者的PBMCs用含50 ng/mL佛波醇12-肉豆蔻酸酯13-乙酸酯与1 μg/mL离子霉素的细胞刺激鸡尾酒（P/I；eBioscience）激活；其二，将4名供者的PBMCs在Dynabeads Human T-Activator CD3/CD28（Gibco）存在的条件下培养。细胞因子浓度采用标准LEGENDplex™人炎症因子检测面板1（BioLegend®）、LSRFortessa（BD）及LEGENDplex™软件（版本2022-02-10，Qognit公司）进行测定。 细胞活力检测实验使用3名供者的PBMCs，细胞因子抑制实验则使用4名供者的PBMCs。 化合物详情及各数据集的变量信息详见《Compound details and Abbreviations.xlsx》文件。各实验的统计分析与绘图所用的R脚本已分别提供。