School of Pharmacy, Changchun University of Chinese Medicine, Changchun, 130117, China.
Jilin Yimintang Pharmaceutical Co., Ltd, Siping, 136000, China.
J Ethnopharmacol. 2024 Dec 5;335:118643. doi: 10.1016/j.jep.2024.118643. Epub 2024 Jul 31.
Feining keli (FNKL) is herbal preparation mainly made from Senecio cannabifolius Less., In recent years, more and more studies have found that FNKL has excellent therapeutic effects on chronic bronchitis (CB). Nevertheless, its pharmacodynamic material basis and mechanism of action are still unknown.
This study aimed to explore the pharmacodynamic material basis and mechanism of action of FNKL in treating CB.
The CB rat model was induced using nasal drops of lipopolysaccharide (LPS) in combination with smoking. Various assessments including behavioral and body mass examination, lung index measurement, enzyme linked immunosorbent assay (ELISA), as well as histological analyses using hematoxylin and eosin (H&E) and Masson staining were conducted to validate the reliability of the CB model. The serum components of FNKL in CB rats were identified using ultra-high-performance liquid chromatography Orbitrap Exploris mass spectrometer (UHPLC-OE-MS). Network pharmacology was used to predict the network of action of the active ingredients in FNKL based on these serum components. Signaling pathways were enriched and analyzed, and molecular docking was conducted for key targets. Molecular dynamics simulations were performed using GROMACS software. The mechanism was confirmed through a series of experiments including Western blot (WB), immunofluorescence (IF), and reverse transcription (RT)-PCR. Additionally, untargeted metabolomics was employed to identify biomarkers and relevant metabolic pathways associated with the treatment of CB with FNKL.
In CB rats, FNKL improved body mass, lung index, and pathological damage of lung tissues. It also decreased interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), malonaldehyde (MDA) levels, and percentage of lung collagen fiber area. Furthermore, FNKL increased IL-10 and superoxide dismutase (SOD) levels, which helped alleviate bronchial inflammation in the lungs. A total of 70 FNKL chemical components were identified in CB rat serum. Through network pharmacology analysis, 5 targets, such as PI3K, AKT, NF-κB, HIF-1α, and MYD88, were identified as key targets of FNKL in the treatment of CB. Additionally, the key signaling pathways identified were PI3K/AKT pathway、NF-κB/MyD88 pathway、HIF-1α pathway. WB, IF, and RT-PCR experiments were conducted to confirm the findings. Molecular docking studies demonstrated successful docking of 16 potential active components with 5 key targets. Additionally, molecular dynamics simulations indicated the stability of quercetin-3-galactoside and HIF-1α. Metabolomics analysis revealed that FNKL primarily regulated pathways related to alpha-linolenic acid metabolism, primary bile acid biosynthesis, bile secretion, arachidonic acid metabolism, neuroactive ligand-receptor interaction, and folate biosynthesis. Furthermore, the expression levels of traumatic acid, traumatin, alpha linolenic acid, cholic acid, 2-arachidonoylglycerol, deoxycholic acid, 7,8-dihydroneopterin, and other metabolites were found to be regulated.
FNKL exhibits positive therapeutic effects on CB, with quercetin-3-galactoside identified as a key active component. The mechanism of FNKL's therapeutic action on CB involves reducing inflammatory response, oxidative stress, and regulating metabolism, and its molecular mechanism was better elucidated in a holistic manner. This study serves as a reference for understanding the pharmacodynamic material basis and mechanism of action of FNKL in treating CB, and provides avenues for exploring the effects of compounded herbal medicines on CB.
肺宁颗粒（FNKL）主要由千里光（Senecio cannabifolius Less.）制成的草药制剂。近年来，越来越多的研究发现 FNKL 对慢性支气管炎（CB）具有极好的治疗作用。然而，其药效物质基础和作用机制尚不清楚。
本研究旨在探讨 FNKL 治疗 CB 的药效物质基础和作用机制。
采用脂多糖（LPS）滴鼻联合吸烟的方法诱导 CB 大鼠模型。通过行为和体重检查、肺指数测量、酶联免疫吸附测定（ELISA）以及苏木精和伊红（H&E）和 Masson 染色的组织学分析等各种评估方法，验证 CB 模型的可靠性。使用超高效液相色谱 Orbitrap 探索质谱仪（UHPLC-OE-MS）鉴定 FNKL 在 CB 大鼠血清中的成分。基于这些血清成分，采用网络药理学预测 FNKL 中活性成分的作用网络。对信号通路进行富集和分析，并进行关键靶标的分子对接。使用 GROMACS 软件进行分子动力学模拟。通过 Western blot（WB）、免疫荧光（IF）和逆转录（RT）-PCR 等一系列实验对机制进行验证。此外，采用非靶向代谢组学方法鉴定与 FNKL 治疗 CB 相关的生物标志物和相关代谢途径。
在 CB 大鼠中，FNKL 改善了体重、肺指数和肺组织的病理损伤。它还降低了白细胞介素（IL）-6、肿瘤坏死因子-α（TNF-α）、丙二醛（MDA）水平和肺胶原纤维面积的百分比。此外，FNKL 增加了 IL-10 和超氧化物歧化酶（SOD）水平，有助于缓解肺部的支气管炎症。在 CB 大鼠血清中鉴定出 70 种 FNKL 化学物质。通过网络药理学分析，鉴定出 5 个关键靶标，如 PI3K、AKT、NF-κB、HIF-1α和 MYD88，作为 FNKL 治疗 CB 的关键靶标。此外，还确定了 PI3K/AKT 途径、NF-κB/MyD88 途径、HIF-1α途径等关键信号通路。WB、IF 和 RT-PCR 实验用于验证这些发现。分子对接研究表明，16 种潜在的活性成分与 5 个关键靶标成功对接。此外，分子动力学模拟表明槲皮素-3-半乳糖苷和 HIF-1α的稳定性。代谢组学分析表明，FNKL 主要调节与α-亚麻酸代谢、初级胆汁酸生物合成、胆汁分泌、花生四烯酸代谢、神经活性配体-受体相互作用和叶酸生物合成相关的途径。此外，还发现调节创伤酸、traumatin、α-亚麻酸、胆酸、2-花生四烯酰甘油、脱氧胆酸、7,8-二氢新喋呤和其他代谢物的表达水平。
FNKL 对 CB 具有积极的治疗作用，槲皮素-3-半乳糖苷被确定为关键的活性成分。FNKL 治疗 CB 的作用机制涉及减轻炎症反应、氧化应激和调节代谢，并且以整体方式更好地阐明了其分子机制。本研究为了解 FNKL 治疗 CB 的药效物质基础和作用机制提供了参考，并为探索复方草药对 CB 的作用提供了途径。
