Wang Ya-Zhu, Wang Xiao-Ting, Tian Shu-Qing, Gao Hui
School of Pharmacy, Liaoning University of Traditional Chinese Medicine Dalian 116600, China Traditional Chinese Medicine Processing Technology Inheritance Base (Liaoning) of the National Administration of Traditional Chinese Medicine Dalian 116600, China.
Zhongguo Zhong Yao Za Zhi. 2024 Oct;49(20):5460-5468. doi: 10.19540/j.cnki.cjcmm.20240716.304.
This study employed network pharmacology, molecular docking, and animal experiments to investigate the mechanism of ginger-processed Anemarrhenae Rhizoma in treating cough due to cold and dampness in lung, aiming to provide a modern pharmacological basis for this therapy. The active ingredients of ginger-processed Anemarrhenae Rhizoma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP) with the oral bioavailability(OB≥30%) and drug likeness(DL≥0.18) as the screening conditions and from the relevant literature. A library of the active ingredients of ginger-processed Anemarrhenae Rhizoma was established, and SwissTargetPrediction was used to predict the potential targets of the active ingredients. The western medical symptom corresponding to cough due to cold and dampness in lung was identified as acute bronchitis based on SymMap, and the targets related to acute bronchitis were obtained from GeneCards, and the Online Mendelian Inheritance in Man(OMIM). Venny was utilized to obtain the common targets between the active ingredients and the disease, and STRING and Cytoscape were used to establish the protein-protein interaction(PPI) network for the common targets. The Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses of potential therapeutic targets were performed with DAVID. AutoDock was used to simulate the molecular docking between the active ingredients with potential targets. Finally, a mouse model of acute bronchitis was prepared with the smoke plus cold stimulation method for validation. In this study, a total of 557 potential targets of the active ingredients of ginger-processed Anemarrhenae Rhizoma were retrieved, including 157 common targets shared with acute bronchitis. The enrichment analysis yielded 1 072 GO terms and 169 KEGG pathways. The results of molecular docking showed that the active ingredients had good binding affinity with MAPK14 and MMP3, and timosaponin AⅢ showcased the strongest binding affinity among the active ingredients. In the animal experiments, other treatment groups except the low-and high-dose raw Anemarrhenae Rhizoma and the ginger decoction groups showed reduced pathological changes in the lung tissue, attenuated inflammatory cell infiltration, lowered levels of interleukin(IL)-1β, IL-18, and tumor necrosis factor(TNF)-α in the bronchoalveolar lavage fluid(BALF), and down-regulated mRNA and protein levels of MAPK14 and MMP3 in the lung tissue. Moreover, ginger-processed Anemarrhenae Rhizoma outperformed raw Anemarrhenae Rhizoma. This study preliminarily revealed that ginger-processed Anemarrhenae Rhizoma can inhibit inflammation by regulating the IL-17, TNF, and other signaling pathways to treat cough due to cold and dampness in lung, verifying the rationality of this therapy. The findings provide a basis for the inheritance and application of the characteristic ginger processing of Anemarrhenae Rhizoma.
本研究采用网络药理学、分子对接和动物实验等方法,探讨姜制知母治疗肺寒痰湿咳嗽的作用机制,旨在为该疗法提供现代药理学依据。以口服生物利用度(OB≥30%)和类药性(DL≥0.18)为筛选条件,从中药系统药理学数据库与分析平台(TCMSP)及相关文献中检索姜制知母的活性成分。建立姜制知母活性成分库,利用SwissTargetPrediction预测活性成分的潜在靶点。基于SymMap将肺寒痰湿咳嗽对应的西医症状确定为急性支气管炎,从GeneCards和《人类孟德尔遗传在线》(OMIM)获取与急性支气管炎相关的靶点。利用Venny获取活性成分与疾病之间的共同靶点,运用STRING和Cytoscape构建共同靶点的蛋白质-蛋白质相互作用(PPI)网络。使用DAVID对潜在治疗靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)通路富集分析。采用AutoDock模拟活性成分与潜在靶点之间的分子对接。最后,采用烟熏加冷刺激法制备急性支气管炎小鼠模型进行验证。本研究共检索到姜制知母活性成分潜在靶点557个,其中与急性支气管炎共有的靶点157个。富集分析得到1072个GO术语和169条KEGG通路。分子对接结果显示,活性成分与MAPK14和MMP3具有良好的结合亲和力,知母皂苷AⅢ在活性成分中表现出最强的结合亲和力。在动物实验中,除低、高剂量生知母组和生姜水煎液组外,其他治疗组肺组织病理变化减轻,炎症细胞浸润减弱,支气管肺泡灌洗液(BALF)中白细胞介素(IL)-1β、IL-18和肿瘤坏死因子(TNF)-α水平降低,肺组织中MAPK14和MMP3的mRNA和蛋白水平下调。此外,姜制知母的效果优于生知母。本研究初步揭示,姜制知母可通过调节IL-17、TNF等信号通路抑制炎症,从而治疗肺寒痰湿咳嗽,验证了该疗法的合理性。研究结果为知母特色姜制工艺的传承与应用提供了依据。