Zheng Yanjun, Song Bei, Lin Jingrong, Ma Jian
Shanghai Key Laboratory of Molecular Imaging, Zhoupu Hospital, Shanghai University of Medicine and Health Sciences, Shanghai 201318, China.
Department of General Practice, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Int Immunopharmacol. 2025 Aug 28;161:115033. doi: 10.1016/j.intimp.2025.115033. Epub 2025 Jun 8.
Sepsis is a life-threatening condition characterized by an excessive inflammatory response and immune dysregulation. Effective treatments remain limited, necessitating the exploration of novel therapeutic agents. Although berbamine (BBM) exhibits notable anti-inflammatory properties, its specific mechanisms and therapeutic potential in sepsis have yet to be elucidated.
This study aims to investigate the protective effects of BBM in sepsis models and elucidate its underlying mechanisms, with a focus on immune modulation and the Notch1 signaling pathway.
We employed two murine sepsis models-lipopolysaccharide (LPS)-induced endotoxemia and cecal ligation and puncture (CLP)-induced sepsis-to assess BBM's therapeutic potential. Histopathological and cytokine analyses were performed to evaluate inflammation and lung injury. Flow cytometry was used to examine immune cell development. Additionally, in vitro assays with bone marrow-derived macrophages (BMDMs) were conducted to assess cytokine production. Network pharmacology and cellular thermal shift assays (CETSA) were utilized to identify potential molecular targets, focusing on Notch1 signaling. We also evaluated the effects of BBM on myeloid-specific Notch1 knockout (Notch1) mice.
BBM significantly improved survival rates and reduced lung inflammation in septic mice. In vitro, BBM inhibited LPS-induced secretion of proinflammatory cytokines (TNF-α, IL-6, and IL-1β) in BMDMs without impairing macrophage differentiation or viability. Network pharmacology analyses identified Notch1 as a potential BBM target, which was validated by CETSA. Mechanistically, BBM accelerated Notch1 degradation via the ubiquitin-proteasome pathway, leading to decreased Notch1 protein levels. Notably, notch1-deficient macrophages exhibited blunted inflammatory responses to LPS, and BBM treatment failed to confer additional suppression, indicating that BBM's anti-septic effects are mediated predominantly through Notch1 inhibition.
BBM exerts a protective effect in sepsis by suppressing inflammation and promoting Notch1 degradation. These findings suggest that BBM may serve as a promising therapeutic agent for sepsis.
脓毒症是一种危及生命的病症，其特征为过度的炎症反应和免疫失调。有效的治疗方法仍然有限，因此需要探索新型治疗药物。尽管小檗胺（BBM）具有显著的抗炎特性，但其在脓毒症中的具体机制和治疗潜力尚未阐明。
本研究旨在探讨BBM在脓毒症模型中的保护作用，并阐明其潜在机制，重点关注免疫调节和Notch1信号通路。
我们采用两种小鼠脓毒症模型——脂多糖（LPS）诱导的内毒素血症和盲肠结扎穿孔（CLP）诱导的脓毒症——来评估BBM的治疗潜力。进行组织病理学和细胞因子分析以评估炎症和肺损伤。使用流式细胞术检测免疫细胞发育。此外，对骨髓来源的巨噬细胞（BMDM）进行体外试验以评估细胞因子产生。利用网络药理学和细胞热位移分析（CETSA）来确定潜在的分子靶点，重点是Notch1信号通路。我们还评估了BBM对髓系特异性Notch1基因敲除（Notch1）小鼠的影响。
BBM显著提高了脓毒症小鼠的存活率并减轻了肺部炎症。在体外，BBM抑制LPS诱导的BMDM中促炎细胞因子（TNF-α、IL-6和IL-1β）的分泌，而不损害巨噬细胞的分化或活力。网络药理学分析确定Notch1为BBM的潜在靶点，这通过CETSA得到验证。机制上，BBM通过泛素-蛋白酶体途径加速Notch1降解，导致Notch1蛋白水平降低。值得注意的是，Notch1缺陷型巨噬细胞对LPS的炎症反应减弱，BBM治疗未能提供额外的抑制作用，表明BBM的抗脓毒症作用主要通过Notch1抑制介导。
BBM通过抑制炎症和促进Notch1降解在脓毒症中发挥保护作用。这些发现表明BBM可能是一种有前景脓毒症治疗药物。
