Table1_Penilloic acid is the chief culprit involved in non-IgE mediated, immediate penicillin-induced hypersensitivity reactions in mice.pdf

Metabolites/impurities (MIs) of penicillin are normally considered to be the main substances inducing immediate hypersensitivity reactions in penicillin treatment. Our previous research found that penicillin can cause non-allergic hypersensitivity reactions (NAHRs) by directly triggering vascular hyperpermeability and exudative inflammation. However, the chief culprits and underlying mechanisms involved in penicillin-induced NAHRs have not yet been fully elucidated. In this study, we used a combination of approaches including a mouse non-allergic hypersensitivity reaction model, UPLC-MS/MS analyses of arachidonic acid metabolites (AAMs), immunoblotting technique, and molecular docking, etc to investigate the culprits involved in penicillin-induced hypersensitivity reactions. We found penilloic acid, one of the main MIs of penicillin, could trigger NAHRs via inducing increased vascular permeability, while the other MIs did no exhibit similar effect. Penilloic acid-induced reactions were not IgE-dependent. Significantly increased arachidonic acids and cascade metabolites in lungs, and activation of RhoA/ROCK signaling pathway in the ears and lungs of mice were noticed after once administration of penilloic acid. This study revealed that penilloic acid was the chief culprit involved in penicillin-induced immediate NAHRs in mice, which mainly associated with direct stimulation of vascular hyperpermeability and exudative inflammation. The activations of AAMs and RhoA/ROCK signaling pathway played important roles in these reactions.

青霉素的代谢物/杂质（Metabolites/impurities，简称MIs）通常被认为是青霉素治疗过程中诱发速发型超敏反应的主要物质。我们既往的研究发现，青霉素可通过直接触发血管通透性增高与渗出性炎症，引发非变应性超敏反应（non-allergic hypersensitivity reactions，简称NAHRs）。然而，青霉素诱导NAHRs的关键致病因子与潜在分子机制尚未完全阐明。本研究结合小鼠非变应性超敏反应模型、花生四烯酸代谢物（arachidonic acid metabolites，简称AAMs）的超高效液相色谱-串联质谱（UPLC-MS/MS）分析、免疫印迹（immunoblotting）技术及分子对接（molecular docking）等多种实验手段，探究青霉素诱导超敏反应的关键致病因子。研究发现，青霉素主要MIs之一的青霉酸（penilloic acid）可通过诱导血管通透性增高触发NAHRs，而其余代谢物/杂质未表现出类似作用。青霉酸诱导的反应不依赖免疫球蛋白E（IgE）。单次给予青霉酸后，小鼠肺部的花生四烯酸及其级联代谢物水平显著升高，耳部与肺部的RhoA/ROCK信号通路（RhoA/ROCK signaling pathway）被激活。本研究揭示，青霉酸是小鼠体内青霉素诱导的速发型NAHRs的关键致病因子，其致病机制主要与直接刺激血管通透性增高及渗出性炎症相关；花生四烯酸代谢物与RhoA/ROCK信号通路的激活在该反应过程中发挥了重要作用。