Objectives: Sulforaphane, a major ingredient isolated from Brassica oleracea var. italica (broccoli), is known to exhibit anti-inflammatory, anti-cancer, and anti-diabetic effects. In this study, we employed an in vitro model of phorbol 12-myristate 13-acetate and a23187 (PMACI)-stimulated human mast cells (HMC-1 cells) to investigate the anti-allergic inflammatory effects and mechanisms of sulforaphane and Brassica oleracea var. italica extracts.Methods: Cytokine levels were measured by ELISA and quantitative real-time-PCR methods. Caspase-1 activity was determined by caspase-1 assay. Binding mode of sulforaphane within caspase-1 was determined by molecular docking simulation. Protein expression was determined by Western blotting.Results: Water extract of Brassica oleracea var. italica (WE) significantly reduced thymic stromal lymphopoietin (TSLP) secretion and caspase-1 activity on activated HMC-1 cells. In the molecular docking simulation and in vitro caspase-1 assays, sulforaphane regulated caspase-1 activity by docking with the identical binding site of caspase-1. Sulforaphane significantly inhibited the levels of inflammatory mediators including TSLP, tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-8 in a dose-dependent manner. Immunoblotting experiments revealed that sulforaphane and WE reduced translocation of NF-κBp65 into the nucleus and phosphorylation of IκBα in the cytosol. Furthermore, phosphorylation of mitogen-activated protein kinases (MAPK) was down-regulated by treatment with sulforaphane or WE.Conclusion: Our findings suggest that sulforaphane and WE have anti-allergic inflammatory effects by intercepting caspase-1/NF-κB/MAPKs signaling pathways.

中文翻译：

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萝卜硫素在计算机模拟和体外模型中减轻了肥大细胞介导的过敏性炎症反应。

目的：萝卜硫素，一种从芸苔属植物中分离出来的主要成分。italica（西兰花）已知具有消炎，抗癌和抗糖尿病作用。在这项研究中，我们采用了佛波醇12-肉豆蔻酸13-乙酸和a23187（PMACI）刺激的人类肥大细胞（HMC-1细胞）的体外模型，研究了萝卜硫素和甘蓝型油菜的抗过敏炎症作用及其​​机制。方法：采用ELISA法和实时荧光定量PCR法检测细胞因子水平。Caspase-1活性通过caspase-1测定来确定。通过分子对接模拟确定了萝卜硫素在caspase-1中的结合方式。通过蛋白质印迹法测定蛋白质表达。结果：芸苔水提取物。italica（WE）显着降低了胸腺基质淋巴细胞生成素（TSLP）的分泌和活化HMC-1细胞上caspase-1的活性。在分子对接模拟和体外caspase-1分析中，萝卜硫素通过与caspase-1的相同结合位点对接来调节caspase-1活性。萝卜硫烷以剂量依赖的方式显着抑制包括TSLP，肿瘤坏死因子（TNF）-α，白介素（IL）-1β，IL-6和IL-8在内的炎症介质的水平。免疫印迹实验表明，萝卜硫烷和WE减少了NF-κBp65进入细胞核的转运和细胞质中IκBα的磷酸化。此外，通过萝卜硫烷或WE的处理下调了促分裂原活化蛋白激酶（MAPK）的磷酸化。