ALI is characterized by macrophage-driven inflammation, causing severe lung damage. Currently, there are limited therapeutic options available for ALI. Liensinine (LIEN), with known anti-inflammatory properties, lacks extensive study in the ALI context. This study aimed to investigate the impact of LIEN on ALI and elucidate its molecular mechanisms. A total of thirty-six male BALB/c mice altogether were split into six groups: Control, LPS (10 mg/kg), Low (10 mg/kg LIEN + 10 mg/kg LPS), Middle (20 mg/kg LIEN + 10 mg/kg LPS), High (40 mg/kg LIEN + 10 mg/kg LPS), and DEX (2 mg/kg DEX + 10 mg/kg LPS). Lung tissue injury, pulmonary edema, and inflammatory factor levels were evaluated in lung tissues and LPS-stimulated bone marrow-derived macrophages (BMDM). TAK1 activation, TRAF6 ubiquitination, and their interactions were assessed to understand the involved molecular mechanisms. LIEN treatment ameliorated lung tissue injury and suppressed LPS-induced inflammatory factor levels in lung tissues and BMDM. Mechanistically, LIEN inhibited TAK1 activation by disrupting TRAF6–TAK1 interactions, limiting p65's nuclear translocation, and reducing the release of inflammatory factors. According to network pharmacology and molecular docking, LIEN most likely prevents inflammation by interfering directly with the Src. Overexpression of Src in BMDM abolished the regulation of TRAF6 by LIEN, supporting the involvement of the Src/TRAF6/TAK1 axis in its mechanism of action. Based on this study, LIEN treats ALI by modifying the Src/TRAF6/TAK1 axis and blocking the activation of the NF-κB pathway, regulating the release of inflammatory factors. These findings highlight the promise of LIEN as a prospective therapeutic option for the treatment of ALI.

ALI 的特点是巨噬细胞驱动的炎症，导致严重的肺部损伤。目前，ALI 的治疗选择有限。莲心碱 (LIEN) 具有已知的抗炎特性，但缺乏针对 ALI 的广泛研究。本研究旨在探讨LIEN对ALI的影响并阐明其分子机制。总共 36 只雄性 BALB/c 小鼠分为六组：对照组、LPS（10 mg/kg）、低（10 mg/kg LIEN + 10 mg/kg LPS）、中（20 mg/kg LIEN） + 10 mg/kg LPS）、高（40 mg/kg LIEN + 10 mg/kg LPS）和 DEX（2 mg/kg DEX + 10 mg/kg LPS）。在肺组织和 LPS 刺激的骨髓源性巨噬细胞 (BMDM) 中评估肺组织损伤、肺水肿和炎症因子水平。评估 TAK1 激活、TRAF6 泛素化及其相互作用，以了解所涉及的分子机制。LIEN 治疗可改善肺组织损伤并抑制肺组织和 BMDM 中 LPS 诱导的炎症因子水平。从机制上讲，LIEN 通过破坏 TRAF6-TAK1 相互作用、限制 p65 核转位和减少炎症因子的释放来抑制 TAK1 激活。根据网络药理学和分子对接，LIEN很可能通过直接干扰Src来预防炎症。BMDM 中 Src 的过表达消除了 LIEN 对 TRAF6 的调节，支持 Src/TRAF6/TAK1 轴参与其作用机制。基于这项研究，LIEN通过修饰Src/TRAF6/TAK1轴并阻断NF-κB通路的激活，调节炎症因子的释放来治疗ALI。这些发现凸显了 LIEN 作为治疗 ALI 的前瞻性治疗选择的前景。