■食物过敏在美国是一种普遍的疾病,影响近3000万人。这种情况的主要管理策略是避免食物,有限的治疗选择是可用的。病理性IgE和过度反应性肥大细胞/嗜碱性粒细胞的升高是食物过敏过敏反应的核心因素。本研究旨在全面评估一种名为formonetin的小分子化合物在调节IgE和肥大细胞活化方面的潜在治疗机制。
■在这项研究中,我们使用ELISA测定了芒柄花素对食物过敏患者外周血单核细胞产生人IgE的抑制作用.我们还使用β-己糖胺酶测定法测量了海蒙素对RBL-2H3和KU812细胞中肥大细胞脱颗粒的预防作用。目的:确定在IgE介导的疾病中可能的靶点,肥大细胞疾病,食物过敏,我们利用计算模型来分析来自各种数据库的formononetin的机械目标,包括SEA,瑞士目标预测,PubChem,基因卡,还有MalaCards.我们产生了KEGG通路,基因本体论,和使用这些靶标的复合靶标途径疾病网络。最后,我们使用qRT-PCR检测KU812和U266细胞系中选定靶标的基因表达。
■Formononetin以剂量依赖的方式显着降低了产生IgE的人骨髓瘤细胞和食物过敏患者的PBMC中的IgE产生,而没有细胞毒性。Formononetin降低了RBL-2H3细胞和KU812细胞中β-己糖胺酶的释放。Formononetin调节食物过敏的25个目标,51在IgE疾病中,和19在肥大细胞疾病中。KEGG通路和基因本体论分析的靶点显示,formonetin调控疾病通路,原发性免疫缺陷,爱泼斯坦-巴尔病毒,和癌症的途径。海蒙素调节的生物过程包括B细胞增殖,分化,免疫反应,和激活过程。复合靶通路疾病网络鉴定NFKB1、NFKBIA、STAT1、STAT3、CCND1、TP53、TYK2和CASP8作为最高目标,在很高程度上受海蒙素调控。TP53,STAT3,PTPRC,IL2和CD19被鉴定为主要被Formonoetin靶向的蛋白质。qPCR验证了在U266细胞和KU812细胞中IgE调节的Formononetin分子靶标的基因。在U266细胞中,发现Formononetin显着增加NFKBIA的基因表达,TP53和BCL-2在降低BTKTYK基因表达的同时,CASP8、STAT3、CCND1、STAT1、NFKB1、IL7R。在嗜碱性粒细胞KU812细胞中,formonetin显著增加NFKBIA的基因表达,TP53和BCL-2在降低BTK基因表达的同时,泰克,CASP8、STAT3、CCND1、STAT1、NFKB1、IL7R。
■这些发现全面展示了formononetin调节浆细胞IgE产生和肥大细胞脱颗粒的机制。
UNASSIGNED: Food allergy is a prevalent disease in the U.S., affecting nearly 30 million people. The primary management strategy for this condition is food avoidance, as limited treatment options are available. The elevation of pathologic IgE and over-reactive mast cells/basophils is a central factor in food allergy anaphylaxis. This study aims to comprehensively evaluate the potential therapeutic mechanisms of a small molecule compound called formononetin in regulating IgE and mast cell activation.
UNASSIGNED: In this study, we determined the inhibitory effect of formononetin on the production of human IgE from peripheral blood mononuclear cells of food-allergic patients using ELISA. We also measured formononetin\'s effect on preventing mast cell degranulation in RBL-2H3 and KU812 cells using beta-hexosaminidase assay. To identify potential targets of formononetin in IgE-mediated diseases, mast cell disorders, and food allergies, we utilized computational modeling to analyze mechanistic targets of formononetin from various databases, including SEA, Swiss Target Prediction, PubChem, Gene Cards, and Mala Cards. We generated a KEGG pathway, Gene Ontology, and Compound Target Pathway Disease Network using these targets. Finally, we used qRT-PCR to measure the gene expression of selected targets in KU812 and U266 cell lines.
UNASSIGNED: Formononetin significantly decreased IgE production in IgE-producing human myeloma cells and PBMCs from food-allergic patients in a dose-dependent manner without cytotoxicity. Formononetin decreased beta-hexosaminidase release in RBL-2H3 cells and KU812 cells. Formononetin regulates 25 targets in food allergy, 51 in IgE diseases, and 19 in mast cell diseases. KEGG pathway and gene ontology analysis of targets showed that formononetin regulated disease pathways, primary immunodeficiency, Epstein-Barr Virus, and pathways in cancer. The biological processes regulated by formononetin include B cell proliferation, differentiation, immune response, and activation processes. Compound target pathway disease network identified NFKB1, NFKBIA, STAT1, STAT3, CCND1, TP53, TYK2, and CASP8 as the top targets regulated at a high degree by formononetin. TP53, STAT3, PTPRC, IL2, and CD19 were identified as the proteins mostly targeted by formononetin. qPCR validated genes of Formononetin molecular targets of IgE regulation in U266 cells and KU812 cells. In U266 cells, formononetin was found to significantly increase the gene expression of NFKBIA, TP53, and BCL-2 while decreasing the gene expression of BTK TYK, CASP8, STAT3, CCND1, STAT1, NFKB1, IL7R. In basophils KU812 cells, formononetin significantly increased the gene expression of NFKBIA, TP53, and BCL-2 while decreasing the gene expression of BTK, TYK, CASP8, STAT3, CCND1, STAT1, NFKB1, IL7R.
UNASSIGNED: These findings comprehensively present formononetin\'s mechanisms in regulating IgE production in plasma cells and degranulation in mast cells.