Jo Hyein, Kim Misun, Jeoung Jaewhoon, Kim Wonho, Park Yoon Ho, Jung Hyun Suk, Lee Wook, Jeoung Dooil
Department of Biochemistry, Kangwon National University, Chuncheon 24341, Republic of Korea.
Molecules. 2025 Feb 11;30(4):840. doi: 10.3390/molecules30040840.
Rocaglamide (Roc-A), a natural phytochemical isolated from Aglaia species, is known to exert anticancer effects. Allergic inflammation can enhance the tumorigenic potential of cancer cells. We hypothesized that Roc-A could regulate allergic inflammation. Roc-A prevented an antigen from increasing the hallmarks of allergic reactions in vitro. Roc-A suppressed passive cutaneous anaphylaxis (PCA) and passive systemic anaphylaxis (PSA). RNA sequencing analysis showed that Roc-A prevented the antigen from increasing the expression of IL-4 in RBL2H3 cells. Roc-A also prevented the antigen from increasing the expression of interleukin-4 receptor (IL-4R). Roc-A was found to form a hydrogen-bonding network with residues N92 and L64 of IL-4R in a molecular docking simulation. Roc-A prevented the antigen from inducing the binding of IL-4R to JAK1. Chromatin immunoprecipitation (ChIP) assays showed that C-Jun could bind to promoter sequences of IL-4 and IL-4R. Mouse recombinant IL-4 protein increased β-hexosaminidase activity, IL-4R expression, and the hallmarks of allergic inflammation in the antigen-independent manner. Mouse recombinant IL-4 protein increased the expressions of CD163 and arghinase-1 and markers of M2 macrophages, but decreased the expression of iNOS, a marker of M1 macrophages in lung macrophages. Roc-A regulated the effects of a culture medium of antigen-stimulated RBL2H3 cells on the expressions of iNOS and arginase-1 in RAW264.7 macrophages. The blocking of IL-4 or downregulation of IL-4R exerted negative effects on the hallmarks of allergic reactions in vitro. The blocking of IL-4 or downregulation of IL-4R also exerted negative effects on PCA, and the downregulation of IL-4R exerted negative effects on PSA. An miR-34a mimic exerted negative effects on allergic reactions in vitro. The downregulation of IL-4R prevented the antigen from decreasing the expression of miR-34a in RBL2H3 cells. We identified chemicals that could bind to IL-4R via molecular docking analysis. The IL-4R docking chemical 1536801 prevented the antigen from increasing β-hexosaminidase activity and the hallmarks of allergic reactions. The IL-4R docking chemical 1536801 also exerted a negative effect on PCA. TargetScan analysis predicted miR-34a as a negative regulator of IL-4R. We found that the anti-allergic effect of Roc-A and its mechanisms were associated with miR-34a. Taken together, our results show that understanding IL-4R-mediated allergic reactions can provide clues for the development of anti-allergy therapeutics.
罗卡酰胺（Roc-A）是从米仔兰属植物中分离出的一种天然植物化学物质，已知具有抗癌作用。过敏性炎症可增强癌细胞的致瘤潜能。我们推测Roc-A可能调节过敏性炎症。Roc-A在体外可阻止抗原增加过敏反应的特征。Roc-A可抑制被动皮肤过敏反应（PCA）和被动全身过敏反应（PSA）。RNA测序分析表明，Roc-A可阻止抗原增加RBL2H3细胞中白细胞介素-4（IL-4）的表达。Roc-A还可阻止抗原增加白细胞介素-4受体（IL-4R）的表达。在分子对接模拟中发现，Roc-A与IL-4R的N92和L64残基形成氢键网络。Roc-A可阻止抗原诱导IL-4R与Janus激酶1（JAK1）结合。染色质免疫沉淀（ChIP）分析表明，c-Jun可与IL-4和IL-4R的启动子序列结合。小鼠重组IL-4蛋白以抗原非依赖的方式增加β-己糖胺酶活性、IL-4R表达和过敏性炎症的特征。小鼠重组IL-4蛋白增加了CD163和精氨酸酶-1的表达以及M2巨噬细胞的标志物，但降低了肺巨噬细胞中M1巨噬细胞标志物诱导型一氧化氮合酶（iNOS）的表达。Roc-A调节抗原刺激的RBL2H3细胞培养基对RAW264.7巨噬细胞中iNOS和精氨酸酶-1表达的影响。阻断IL-4或下调IL-4R对体外过敏反应的特征产生负面影响。阻断IL-4或下调IL-4R对PCA也产生负面影响，而下调IL-4R对PSA产生负面影响。miR-34a模拟物对体外过敏反应产生负面影响。下调IL-4R可阻止抗原降低RBL2H3细胞中miR-34a的表达。我们通过分子对接分析鉴定了可与IL-4R结合的化学物质。IL-4R对接化学物质1536801可阻止抗原增加β-己糖胺酶活性和过敏反应的特征。IL-4R对接化学物质1536801对PCA也有负面影响。TargetScan分析预测miR-34a是IL-4R的负调节因子。我们发现Roc-A的抗过敏作用及其机制与miR-34a有关。综上所述，我们的结果表明，了解IL-4R介导的过敏反应可为抗过敏治疗药物的开发提供线索。
