【摘要】 近年来，食品过敏的发生率在全球范围内呈持续上升趋势，已成为免疫学与食品科学交叉领域的研究热点。为探究功能性小分子乳酸在类肥大细胞脱颗粒反应中的作用机制，采用大鼠细胞系RBL-2H3建立C48/80诱导的脱颗粒模型，模拟不同乳酸处理条件对细胞功能的影响。通过细胞活力检测、β-己糖胺酶释放测定、酶联免疫吸附测定、蛋白免疫印迹、实时荧光定量聚合酶链式反应和活性氧检测等手段，系统评估乳酸对脱颗粒、炎症因子分泌、乳酰化修饰和氧化应激的影响。20 mmol/L的乳酸处理可协同C48/80显著促进细胞脱颗粒，增强组胺和相关炎症因子分泌，并提高乳酰化水平和活性氧积聚。同时，乳酸联合处理会显著上调氧化应激相关基因SLC7A11表达(P<0.05)，提示乳酸诱导的活性氧积累可能达到其表达激活阈值。本研究为乳酸在过敏性炎症中的免疫代谢机制提供了实验依据。更多还原

【Abstract】 In recent years, the global incidence of food allergies has shown a sustained upward trend, emerging as a prominent research focus at the interface of immunology and food science. In order to investigate the regulatory mechanism of the functional small molecule lactic acid in the degranulation response of mast cell-like cells, a degranulation model was established using the RBL-2H3 cell line induced by compound 48/80. Various lactate treatment conditions were designed to simulate lactate accumulation in inflammatory microenvironments. Cell viability assays, β-hexosaminidase release measurements, enzymelinked immunosorbent assay(ELISA), Western blotting, quantitative real-time polymerase chain reaction(qPCR), and reactive oxygen species(ROS) detection were employed to systematically evaluate the effects of lactate on degranulation, inflammatory cytokine secretion, protein lactylation, and oxidative stress. The results showed that treatment with 20 mmol/L lactic acid significantly enhanced compound 48/80-induced degranulation, increased the secretion of histamine and pro-inflammatory cytokines including interleukin-4(IL-4), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α), and elevated intracellular lactylation levels and ROS accumulation. Notably, combined lactate treatment significantly upregulated the expression of the oxidative stress-related gene solute carrier family 7 member 11(SLC7A11)(P<0.05), suggesting that lactate-induced ROS accumulation may reach a threshold necessary for transcriptional activation. This study provides experimental evidence for the immunometabolic role of lactic acid in allergic inflammation and offers a theoretical basis for its potential impact on immune regulation under food-derived or endogenous metabolic conditions.更多还原