Ning Yujie, Huang Liting, Wang Qin-Qin, Liu Lina, Ni Xinghua, Xie Xiaoyun, Liu Jingyu, Su Qian, Yang Shilin, Yuan Renyikun, Gao Hongwei
Engineering Research Center of Innovative Drugs for Traditional Chinese Medicine and Zhuang & Yao Medicine, Ministry of Education, Nanning, 530200, China.
College of Pharmacy, Guangxi University of Chinese Medicine, Nanning, 530200, China.
Chin Med. 2025 Jul 3;20(1):104. doi: 10.1186/s13020-025-01155-5.
Chronic obstructive pulmonary disease (COPD) is a complex respiratory disorder characterized by persistent respiratory symptoms and progressive airflow limitation. Long-term exposure to harmful particulates and gases causes structural abnormalities in the airways and alveoli, activating NF-κB/MAPK signaling pathways that drive chronic inflammation and tissue remodeling. Key features include an imbalance between proteolytic enzymes and inhibitors mediated by matrix metalloproteinases, and excessive mucus secretion due to mucin overexpression. These factors exacerbate airway obstruction and inflammation, contributing to disease progression. Hederasaponin C (HSC), a triterpenoid saponin with anti-inflammatory properties, shows potential in mitigating COPD-related inflammation, but its precise mechanisms require further investigation.
We investigated the impact of HSC on COPD models induced by CSE + LPS using a comprehensive approach. In vitro studies included Western blotting, qRT-PCR, ELISA, and immunofluorescence to assess key proteins in NF-κB/MAPK signaling pathways, MMP9 and MMP12 expression, and mucin levels (MUC-5AC, MUC-5B). Binding affinity between HSC and TLR4 was evaluated using molecular docking, SPR analysis, and CETSA. DNA methylation at MUC-5B chr11:1243469 position was detected using an Agilent 2100 Bioanalyzer. In vivo, a COPD mouse model induced by cigarette smoke and LPS (CS + LPS) was developed, and HSC treatment effects were evaluated using H&E staining, multiplex immunofluorescence staining, Western blot, and ELISA kits.
HSC significantly inhibited CSE + LPS-induced inflammation by targeting TLR4 and attenuating NF-κB/MAPK signaling pathways overactivation. It also downregulated MMP9, MMP12, MUC-5AC, and MUC-5B expression and suppressed MUC-5B chr11:1243469 position DNA methylation. In vivo, HSC alleviated COPD symptoms in CS + LPS-induced mice, reducing TLR4/NF-κB/MAPK signaling pathways overactivation and smoking-associated factors.
HSC targets TLR4, attenuates NF-κB/MAPK signaling pathways overactivation, reduces MMP9, MMP12, MUC-5AC, and MUC-5B expression, and suppresses MUC-5B chr11:1243469 position DNA methylation. These actions reduce inflammation, restore protease-antiprotease balance, and mitigate excessive mucus secretion, highlighting the promise of HSC as a viable treatment strategy for COPD management.
慢性阻塞性肺疾病（COPD）是一种复杂的呼吸系统疾病，其特征为持续的呼吸道症状和进行性气流受限。长期暴露于有害颗粒和气体中会导致气道和肺泡的结构异常，激活驱动慢性炎症和组织重塑的NF-κB/MAPK信号通路。关键特征包括基质金属蛋白酶介导的蛋白水解酶和抑制剂之间的失衡，以及由于粘蛋白过表达导致的黏液分泌过多。这些因素会加剧气道阻塞和炎症，促进疾病进展。常春藤皂苷元C（HSC）是一种具有抗炎特性的三萜皂苷，在减轻COPD相关炎症方面显示出潜力，但其确切机制需要进一步研究。
我们采用综合方法研究了HSC对由香烟烟雾提取物（CSE）+脂多糖（LPS）诱导的COPD模型的影响。体外研究包括蛋白质免疫印迹法、定量逆转录聚合酶链反应（qRT-PCR）、酶联免疫吸附测定（ELISA）和免疫荧光法，以评估NF-κB/MAPK信号通路中的关键蛋白、基质金属蛋白酶9（MMP9）和基质金属蛋白酶12（MMP12）的表达以及粘蛋白水平（MUC-5AC、MUC-5B）。使用分子对接、表面等离子体共振（SPR）分析和热蛋白质组分析（CETSA）评估HSC与Toll样受体4（TLR4）之间的结合亲和力。使用安捷伦2100生物分析仪检测MUC-5B基因座chr11:1243469位置的DNA甲基化。在体内，建立由香烟烟雾和LPS（CS+LPS）诱导的COPD小鼠模型，并使用苏木精-伊红（H&E）染色、多重免疫荧光染色、蛋白质免疫印迹法和ELISA试剂盒评估HSC的治疗效果。
HSC通过靶向TLR4并减弱NF-κB/MAPK信号通路的过度激活，显著抑制了CSE+LPS诱导的炎症。它还下调了MMP9、MMP12、MUC-5AC和MUC-5B的表达，并抑制了MUC-5B基因座chr11:1243469位置的DNA甲基化。在体内，HSC减轻了CS+LPS诱导的小鼠的COPD症状，减少了TLR4/NF-κB/MAPK信号通路的过度激活和吸烟相关因素。
HSC靶向TLR4，减弱NF-κB/MAPK信号通路的过度激活，降低MMP9、MMP12、MUC-5AC和MUC-5B的表达，并抑制MUC-5B基因座chr11:1243469位置的DNA甲基化。这些作用减轻了炎症，恢复了蛋白酶-抗蛋白酶平衡，并减轻了黏液分泌过多，突出了HSC作为一种可行的COPD治疗策略的前景。
