与近几十年全球饮食乳化剂的广泛使用相一致，许多与肠道屏障功能障碍相关的慢性疾病的发病率也在上升。因此，我们对常用的食品乳化剂对胃肠上皮细胞的细胞毒性、屏障功能、转录组变化和蛋白质表达的影响进行了研究。使用诱导多能干细胞生成的人类肠道类器官、结肠器官芯片和液液界面细胞培养在存在两种常见乳化剂：聚山梨酸酯20（P20）和聚山梨酸酯80（P80）的条件下。测量了细胞的细胞毒性、跨上皮电阻（TEER）和细胞间通透性的流量。进行上皮紧密连接（TJ）的免疫荧光染色、RNA测序转录组和靶向蛋白质组学等实验。在所有模型中，细胞在0.1%（体积/体积）浓度P20和P80的暴露下开始溶解。上皮屏障破坏与TEER降低、细胞间通透性增加和不规则的TJ免疫染色相关。RNA测序和靶向蛋白质组学分析表明，细胞发育、信号传导、增殖、凋亡、炎症反应和应激反应在0.05%浓度下被上调，这个浓度低于直接细胞毒性。通过分泌多种细胞因子和趋化因子、与其受体的相互作用以及PI3K-Akt和MAPK信号通路，表明了一种促炎反应的特征。P20诱导CXCL5、CXCL10和VEGFA上调，而P80诱导CXCL1、CXCL8（IL-8）、CXCL10和LIF上调。本研究直接提供了食品乳化剂P20和P80对肠道上皮完整性的有害影响的证据。上皮屏障破坏的潜在机制是在1%到0.1%浓度范围内的细胞死亡。即使在0.1%以下的浓度下，这些聚山梨酸酯也会引起促炎反应，暗示其对胃肠健康的有害影响。© 2023 The Authors. Allergy published by European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.

The rising prevalence of many chronic diseases related to gut barrier dysfunction coincides with the increased global usage of dietary emulsifiers in recent decades. We therefore investigated the effect of the frequently used food emulsifiers on cytotoxicity, barrier function, transcriptome alterations, and protein expression in gastrointestinal epithelial cells.Human intestinal organoids originating from induced pluripotent stem cells, colon organoid organ-on-a-chip, and liquid-liquid interface cells were cultured in the presence of two common emulsifiers: polysorbate 20 (P20) and polysorbate 80 (P80). The cytotoxicity, transepithelial electrical resistance (TEER), and paracellular-flux were measured. Immunofluorescence staining of epithelial tight-junctions (TJ), RNA-seq transcriptome, and targeted proteomics were performed.Cells showed lysis in response to P20 and P80 exposure starting at a 0.1% (v/v) concentration across all models. Epithelial barrier disruption correlated with decreased TEER, increased paracellular-flux and irregular TJ immunostaining. RNA-seq and targeted proteomics analyses demonstrated upregulation of cell development, signaling, proliferation, apoptosis, inflammatory response, and response to stress at 0.05%, a concentration lower than direct cell toxicity. A proinflammatory response was characterized by the secretion of several cytokines and chemokines, interaction with their receptors, and PI3K-Akt and MAPK signaling pathways. CXCL5, CXCL10, and VEGFA were upregulated in response to P20 and CXCL1, CXCL8 (IL-8), CXCL10, LIF in response to P80.The present study provides direct evidence on the detrimental effects of food emulsifiers P20 and P80 on intestinal epithelial integrity. The underlying mechanism of epithelial barrier disruption was cell death at concentrations between 1% and 0.1%. Even at concentrations lower than 0.1%, these polysorbates induced a proinflammatory response suggesting a detrimental effect on gastrointestinal health.© 2023 The Authors. Allergy published by European Academy of Allergy and Clinical Immunology and John Wiley & Sons Ltd.