Wang Yixu, Zhang Jin, Xu Sihan, Li Junjie, Liu Wenjuan, Jiang Min, Bai Gang
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, China.
State Key Laboratory of Medicinal Chemical Biology, College of Pharmacy and Tianjin Key Laboratory of Molecular Drug Research, Nankai University, Tianjin, China.
Phytomedicine. 2025 Jul;142:156807. doi: 10.1016/j.phymed.2025.156807. Epub 2025 Apr 26.
Acute lung injury (ALI) is a critical pathophysiological response in various respiratory diseases characterized by alveolar damage and excessive inflammation. It can progress to acute respiratory distress syndrome, with current treatments showing limited efficacy and considerable side effects. Psoralen (Pso), derived from Psoralea corylifolia l., has anti-inflammatory properties, but its role in ALI remains fully elucidated.
This study aimed to investigate the therapeutic effects of Pso on ALI, and to explore its therapeutic targets and mechanisms.
The therapeutic potential of Pso was assessed using a model of ALI induced by lipopolysaccharide (LPS). The direct target was investigated using alkynyl-Psoralen (A-Pso) for chemical proteomic analysis, and a series of molecular biology methods were used to explore the underlying mechanism.
Pso treatment significantly alleviated LPS-induced lung injury in mice, targeted macrophages, and inhibited the LPS-induced activation of macrophages. Target-fishing experiments identified high-mobility group box-1 (HMGB1) as a direct target of Pso in macrophages. FTS and CETSA confirmed Pso binding to HMGB1, and LC-MS/MS analysis indicated a covalent interaction between Pso and Cys106 of HMGB1. Furthermore, Pso covalently targeted Cys106, affecting HMGB1 binding to TLR4 and downregulating the phosphorylation of NF-κB, indicating the inhibition of the TLR4/NF-κB signaling pathway both in macrophages and in lung tissues of ALI mice. These findings suggest that Pso exerts its therapeutic effects by covalently targeting HMGB1 in macrophages and modulating the TLR4/NF-κB signaling pathway.
This study not only found that Pso improves LPS-induced ALI inflammation by targeting macrophages, but also verified that this mechanism of action is mainly caused by Pso covalently targeting Cys106 of HMGB1, inhibiting the HMGB1-TLR4 interaction, and thereby suppressing cytokine storm generation. As the first naturally derived HMGB1 covalent inhibitor with a clear binding site, Pso plays an important role in HMGB1 induced inflammatory diseases and is an active precursor for the development of new HMGB1 covalent inhibitors.
急性肺损伤（ALI）是多种呼吸系统疾病中的一种关键病理生理反应，其特征为肺泡损伤和过度炎症反应。它可进展为急性呼吸窘迫综合征，目前的治疗方法疗效有限且副作用较大。补骨脂素（Pso）源自补骨脂，具有抗炎特性，但其在ALI中的作用尚未完全阐明。
本研究旨在探讨Pso对ALI的治疗作用，并探索其治疗靶点和机制。
使用脂多糖（LPS）诱导的ALI模型评估Pso的治疗潜力。采用炔基补骨脂素（A-Pso）进行化学蛋白质组学分析以研究直接靶点，并使用一系列分子生物学方法探索潜在机制。
Pso治疗显著减轻了LPS诱导的小鼠肺损伤，作用于巨噬细胞，并抑制了LPS诱导的巨噬细胞活化。靶点捕获实验确定高迁移率族蛋白B1（HMGB1）是Pso在巨噬细胞中的直接靶点。FTS和CETSA证实Pso与HMGB1结合，液相色谱-串联质谱（LC-MS/MS）分析表明Pso与HMGB1的半胱氨酸106之间存在共价相互作用。此外，Pso共价靶向半胱氨酸106，影响HMGB1与Toll样受体4（TLR4）的结合，并下调核因子κB（NF-κB）的磷酸化，表明在巨噬细胞和ALI小鼠肺组织中均抑制了TLR4/NF-κB信号通路。这些发现表明，Pso通过在巨噬细胞中共价靶向HMGB1并调节TLR4/NF-κB信号通路发挥其治疗作用。
本研究不仅发现Pso通过作用于巨噬细胞改善LPS诱导的ALI炎症，还证实了这种作用机制主要是由于Pso共价靶向HMGB1的半胱氨酸106，抑制HMGB1-TLR4相互作用，从而抑制细胞因子风暴的产生。作为首个具有明确结合位点的天然来源的HMGB1共价抑制剂，Pso在HMGB1诱导的炎症性疾病中发挥重要作用，是开发新型HMGB1共价抑制剂的活性前体。
