背景:Feiningkeli(FNKL)是主要由SenecioCannerifoliusLess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)有良好的治疗作用。然而,其药效物质基础和作用机制尚不清楚。
目的:本研究旨在探讨FNKL治疗CB的药效物质基础和作用机制。
方法:采用脂多糖(LPS)滴鼻联合吸烟诱导CB大鼠模型。各种评估,包括行为和体重检查,肺指数测量,酶联免疫吸附测定(ELISA),以及使用苏木精和伊红(H&E)和Masson染色进行组织学分析,以验证CB模型的可靠性.使用超高效液相色谱Orbitrap探索质谱(UHPLC-OE-MS)鉴定CB大鼠FNKL的血清成分。网络药理学用于基于这些血清成分预测FNKL中活性成分的作用网络。信号通路进行了丰富和分析,并对关键靶标进行了分子对接。使用GROMACS软件进行分子动力学模拟。该机制通过一系列实验得到证实,包括蛋白质印迹(WB),免疫荧光(IF),和逆转录(RT)-PCR。此外,非靶向代谢组学用于鉴定与用FNKL治疗CB相关的生物标志物和相关代谢途径。
结果:在CB大鼠中,FNKL改善了体重,肺指数,肺组织病理损害。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IL-10和超氧化物歧化酶(SOD)水平,这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化学成分。通过网络药理学分析,5个目标,例如PI3K,AKT,NF-κB,HIF-1α,和MYD88被确定为FNKL治疗CB的关键靶标。此外,鉴定出的关键信号通路为PI3K/AKT通路、NF-κB/MyD88通路、HIF-1α通路。WB,如果,并进行RT-PCR实验以证实这一发现。分子对接研究证实16个潜在活性组分与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。代谢组学分析显示,FNKL主要调节与α-亚麻酸代谢相关的通路,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。此外,创伤性酸的表达水平,创伤,α亚麻酸,胆酸,2-花生四酰基甘油,脱氧胆酸,7,8-二氢蝶呤,和其他代谢物被发现被调节。
结论:FNKL对CB具有积极的治疗作用,其中槲皮素-3-半乳糖苷被确定为关键活性成分。FNKL对CB的治疗作用机制包括减少炎症反应,氧化应激,调节新陈代谢,其分子机制得到了较好的整体阐明。本研究为了解FNKL治疗CB的药效物质基础和作用机制提供参考,并为探讨复方中药对CB的影响提供了途径。
BACKGROUND: 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.
OBJECTIVE: This study aimed to explore the pharmacodynamic material basis and mechanism of action of FNKL in treating CB.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.