Wang Liming, Chen Hongrui, DU Yan, Zhao Peng, Wang Yujie, Tian Yange, Liu Xinguang, Li Jiansheng
Collaborative Innovation Center for Chinese Medicine and Respiratory Diseases Co-constructed by Henan Province and State Ministry of Education, Henan Key Laboratory of Chinese Medicine for Respiratory Disease, Henan University of Chinese Medicine, Zhengzhou 450046, China.
Academy of Chinese Medical Sciences, Henan University of Chinese Medicine, Zhengzhou 450046, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2025 Jul 20;45(7):1409-1422. doi: 10.12122/j.issn.1673-4254.2025.07.07.
To investigate pharmacologically active components of Formula (YZF) and their mechanisms for alleviating airway inflammation in mice with chronic obstructive pulmonary disease (COPD).
Ultra-high-performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry was employed to characterize the chemical components in YZF and YZF-medicated rat serum. A compound-disease target network was constructed based on serum components of YZF to screen the key pathways and targets using enrichment analysis. A mouse model of cigarette smoke-induced COPD was used to evaluate the anti-inflammatory effect of YZF and validate the expression of key proteins in network pharmacology-enriched pathways. Fifty male C57BL/6J mice were randomized equally into control group, COPD model group, high- and low-dose YZF treatment groups, and N-acetylcysteine treatment group. Pulmonary function of the mice was assessed using whole-body plethysmography, and lung histopathology, alveolar structure, and airway remodeling were analyzed using HE staining. The levels of IL-1β, IL-6, and TNF‑α in bronchoalveolar lavage fluid (BALF) were determined with ELISA, and pulmonary expressions of PI3K, Akt, phosphorylated Akt (p-Akt), p65, and phosphorylated p65 (p-p65) were detected using immunohistochemistry.
We identified a total of 156 chemical components (including 26 flavonoids or flavonoid glycosides, 27 alkaloids, and 11 saponins) in YZF and 43 prototype components in medicated rat serum. Network pharmacology revealed 704 YZF-related targets and 1199 COPD-associated targets. Integrated analysis suggested that the anti-COPD effects of YZF were associated with the PI3K-Akt signaling pathway. In mouse models of COPD, YZF treatment significantly increased mean alveolar number and peak expiratory flow (<0.05), reduced mean linear intercept, bronchial wall thickness, lung coefficient, and BALF cytokine levels, and suppressed the expressions of PI3K, Akt, p-Akt, p65, and p-p65 in the lung tissues.
YZF alleviates COPD symptoms and airway inflammation in mice possibly by inhibiting the PI3K/Akt/NF‑κB pathway through its multiple components that interact with multiple targets.
研究益肺方（YZF）的药理活性成分及其缓解慢性阻塞性肺疾病（COPD）小鼠气道炎症的机制。
采用超高效液相色谱-四极杆-轨道阱质谱联用技术对YZF及YZF含药血清中的化学成分进行表征。基于YZF的血清成分构建化合物-疾病靶点网络，通过富集分析筛选关键通路和靶点。采用香烟烟雾诱导的COPD小鼠模型评价YZF的抗炎作用，并验证网络药理学富集通路中关键蛋白的表达。将50只雄性C57BL/6J小鼠随机均分为对照组、COPD模型组、YZF高剂量和低剂量治疗组以及N-乙酰半胱氨酸治疗组。采用全身容积描记法评估小鼠肺功能，用HE染色分析肺组织病理学、肺泡结构和气道重塑。用ELISA法测定支气管肺泡灌洗液（BALF）中IL-1β、IL-6和TNF-α水平，用免疫组织化学法检测肺组织中PI3K、Akt、磷酸化Akt（p-Akt）、p65和磷酸化p65（p-p65）的表达。
我们在YZF中总共鉴定出156种化学成分（包括26种黄酮类或黄酮苷类、27种生物碱和11种皂苷），在含药大鼠血清中鉴定出43种原型成分。网络药理学揭示了704个与YZF相关的靶点和1199个与COPD相关的靶点。综合分析表明，YZF的抗COPD作用与PI3K-Akt信号通路有关。在COPD小鼠模型中，YZF治疗显著增加平均肺泡数和呼气峰流量（<0.05），降低平均线性截距、支气管壁厚度、肺系数和BALF细胞因子水平，并抑制肺组织中PI3K、Akt、p-Akt、p65和p-p65的表达。
YZF可能通过其多种成分与多个靶点相互作用抑制PI3K/Akt/NF-κB通路，从而减轻小鼠COPD症状和气道炎症。
