Cao Tianjiao, Li An-Qing, Zhang Yi, Xie Ting-Ting, Weng Ding-Zhou, Pan Chun-Shui, Yan Li, Sun Kai, Wang Di, Han Jing-Yan, Liu Jian
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, PR China; Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, PR China; Academy of Integration of Chinese and Western Medicine, Peking University Health Science Center, Beijing, PR China; The Key Discipline for Integration of Chinese and Western Basic Medicine (Microcirculation) of the National Administration of Traditional Chinese Medicine, Beijing, PR China; Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing, PR China; State Key Laboratory of Core Technology in Innovative Chinese Medicine, Tianjin, PR China; Beijing Microvascular Institute of Integration of Chinese and Western Medicine, Beijing, PR China; The Institute of Cardiovascular Sciences, School of Basic Medical Sciences, State Key Laboratory of Vascular Homeostasis and Remodeling, Health Science Center, Peking University, Beijing, PR China.
Department of Integration of Chinese and Western Medicine, School of Basic Medical Sciences, Peking University, Beijing, PR China; Tasly Microcirculation Research Center, Peking University Health Science Center, Beijing, PR China; Academy of Integration of Chinese and Western Medicine, Peking University Health Science Center, Beijing, PR China; The Key Discipline for Integration of Chinese and Western Basic Medicine (Microcirculation) of the National Administration of Traditional Chinese Medicine, Beijing, PR China; Key Laboratory of Stasis and Phlegm, State Administration of Traditional Chinese Medicine of the People's Republic of China, Beijing, PR China; State Key Laboratory of Core Technology in Innovative Chinese Medicine, Tianjin, PR China; Beijing Microvascular Institute of Integration of Chinese and Western Medicine, Beijing, PR China.
Phytomedicine. 2025 Apr;139:156432. doi: 10.1016/j.phymed.2025.156432. Epub 2025 Jan 30.
Acute lung injury (ALI) has emerged as a critical illness, with sepsis-related ALI accounting for >80 %. In the context of bacterial infection, damage to the pulmonary microvascular barrier leads to inflammatory cell infiltration and plasma component extravasation into pulmonary interstitium. This disruption impairs gas exchange, resulting in hypoxemia. Norwogonin (NWG), a natural plant flavone, has shown potential anti-inflammatory and antioxidative effects. However, whether it could ameliorate sepsis-related ALI and the potential mechanism remains unknown.
This study aims to investigate the effects and underlying mechanisms of NWG in treating sepsis-related ALI.
Male Wistar rats (200-220 g) were used to establish sepsis-related ALI model via intraperitoneal injection of lipopolysaccharide (LPS). Vital signs and arterial blood gas analysis, HE and immunohistochemistry staining, dynamic visualization of the microcirculatory system to observe FITC-dextran leakage and leukocyte adhesion, ELISA assay of inflammatory cytokines, Evans Blue extravasation, measurement of total protein content in bronchoalveolar lavage fluid, determination of the Wet/Dry weight ratio, Western blot and RT-qPCR analysis were used to evaluate NWG's effects and the potential mechanism. Additionally, we employed network pharmacology and molecular docking to identify and evaluate the interaction between NWG and the key targets of ALI. Surface plasmon resonance and enzyme activity assay were utilized to confirm the direct interaction between NWG and the potential targets.
NWG administration improved the vital signs of LPS-stimulated rats. Exposure to LPS led to deteriorated arterial blood gas analysis, prominent lung morphology destruction, neutrophil and M1 macrophage infiltration, leukocyte adhesion, FITC-dextran leakage, elevated secretion of inflammatory cytokines, and aggravated lung edema. NWG intervention effectively mitigated these changes. Furthermore, NWG suppressed NF-κB/NLRP3 signaling and up-regulated endothelial junction proteins. Network pharmacology analysis and molecular docking identified five top key targets: MMP-9, AKT1, COX-2, Src and JAK-2. Western blot and RT-qPCR results confirmed that NWG inhibited the Src/AKT1/NF-κB signaling pathway, and down-regulated the levels of inflammatory factors. Surface plasmon resonance revealed the direct binding between NWG and AKT1, COX-2 and Src, rather than MMP-9. Enzyme activity assay demonstrated that NWG inhibited the activity of AKT1, COX-2 and Src.
NWG alleviated inflammation, restored pulmonary microvascular barrier function and improved LPS-induced ALI. These effects were mediated by inhibiting the Src/AKT1/NF-κB signaling pathway through direct targeting of Src, AKT1 and COX-2. Our study provided novel scientific evidence supporting the use of NWG in the treatment of ALI caused by sepsis.
急性肺损伤（ALI）已成为一种危重病，其中脓毒症相关的ALI占比超过80%。在细菌感染的情况下，肺微血管屏障受损会导致炎症细胞浸润以及血浆成分渗入肺间质。这种破坏会损害气体交换，导致低氧血症。去甲淫羊藿素（NWG）是一种天然植物黄酮，已显示出潜在的抗炎和抗氧化作用。然而，它是否能改善脓毒症相关的ALI及其潜在机制仍不清楚。
本研究旨在探讨NWG治疗脓毒症相关ALI的作用及潜在机制。
雄性Wistar大鼠（200 - 220 g）通过腹腔注射脂多糖（LPS）建立脓毒症相关ALI模型。采用生命体征和动脉血气分析、HE和免疫组化染色、微循环系统动态可视化观察异硫氰酸荧光素 - 葡聚糖渗漏和白细胞黏附、炎症细胞因子的ELISA检测、伊文思蓝外渗、支气管肺泡灌洗液总蛋白含量测定、湿/干重比测定、蛋白质免疫印迹法和逆转录 - 定量聚合酶链反应分析来评估NWG的作用及潜在机制。此外，我们运用网络药理学和分子对接来识别和评估NWG与ALI关键靶点之间的相互作用。利用表面等离子体共振和酶活性测定来确认NWG与潜在靶点之间的直接相互作用。
给予NWG改善了LPS刺激大鼠的生命体征。LPS暴露导致动脉血气分析恶化、肺部形态破坏显著、中性粒细胞和M1巨噬细胞浸润、白细胞黏附、异硫氰酸荧光素 - 葡聚糖渗漏、炎症细胞因子分泌增加以及肺水肿加重。NWG干预有效减轻了这些变化。此外，NWG抑制NF - κB/NLRP3信号通路并上调内皮连接蛋白。网络药理学分析和分子对接确定了五个关键靶点：基质金属蛋白酶 - 9（MMP - 9）、蛋白激酶B（AKT1）、环氧合酶 - 2（COX - 2）、肉瘤激酶（Src）和Janus激酶2（JAK - 2）。蛋白质免疫印迹法和逆转录 - 定量聚合酶链反应结果证实，NWG抑制Src/AKT1/NF - κB信号通路，并下调炎症因子水平。表面等离子体共振显示NWG与AKT1、COX - 2和Src直接结合，而非与MMP - 9结合。酶活性测定表明NWG抑制AKT1、COX - 2和Src的活性。
NWG减轻炎症，恢复肺微血管屏障功能并改善LPS诱导的ALI。这些作用是通过直接靶向Src、AKT1和COX - 2抑制Src/AKT1/NF - κB信号通路介导的。我们的研究提供了新的科学证据支持NWG用于治疗脓毒症引起的ALI。
