摘要
目的通过网络药理学与分子对接技术探讨固本止咳平喘颗粒治疗咳嗽变异性哮喘(CVA)的分子机制,利用CVA模型大鼠进行验证,为固本止咳平喘颗粒的基础研究及临床应用提供依据。方法通过TCMSP、BATMAN-TCM数据库检索固本止咳平喘颗粒药物活性成分及对应靶点,利用OMIM、GeneCards、DrugBank、PharmGKB数据库筛选CVA疾病靶点,获取药物-疾病交集靶点;使用Cytoscape3.10.1软件构建药物-成分-靶点网络,筛选关键成分;STRING数据库构建蛋白相互作用(PPI)网络,筛选核心靶点;使用DAVID平台进行GO和KEGG通路富集分析;通过AutoDock Vina软件对度值较高的核心靶点和关键活性成分进行分子对接。采用烟熏联合腹腔及皮下多点注射卵清白蛋白建立CVA大鼠模型,将大鼠随机分为空白组、模型组、孟鲁司特钠片组及固本止咳平喘颗粒低、高剂量组,各给药组予相应药物干预15 d,HE染色观察肺组织形态,ELISA检测血清白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量,免疫组化检测肺组织AKT、EGFR、NFKB1阳性表达。结果共筛选出固本止咳平喘颗粒活性成分9996个、潜在靶点1119个,CVA靶点2740个,药物与疾病交集靶点467个;PPI网络拓扑分析显示,AKT1、STAT3、EGFR、NFKB1、MTOR为核心靶点;富集分析结果表明,固本止咳平喘颗粒治疗CVA主要通过EGFR/PI3K-Akt/NF-κB信号通路发挥抗炎作用;分子对接显示核心靶点与关键成分均有较好的结合能力,EGFR与Quercetin的结合能<-7.0 kcal/mol。动物实验结果显示,固本止咳平喘颗粒干预后,模型大鼠肺组织肺泡壁变窄,肺泡大小均一,淋巴细胞浸润明显改善,细支气管收缩,管腔内黏液减少;与模型组比较,固本止咳平喘颗粒低、高剂量组和孟鲁司特钠片组血清IL-1β、TNF-α含量降低,肺组织AKT、EGFR、NFKB1阳性表达下降(P<0.05,P<0.01)。结论固本止咳平喘颗粒可能通过EGFR/PI3K-Akt/NF-κB信号通路介导的炎症反应,显著改善CVA肺组织结构损伤,抑制炎性细胞浸润,下调炎症因子表达,从而发挥治疗CVA作用。
Objective To explore the molecular mechanism of Guben Zhike Pingchuan Granules in treating cough variant asthma(CVA)through network pharmacology and molecular docking technology;To use CVA model rats for validation;To provide a basis for the basic research and clinical application of Guben Zhike Pingchuan Granules.Methods TCMSP and BATMAN-TCM databases were used to retrieve the active components and corresponding targets of Guben Zhike Pingchuan Granules,and CVA disease targets were screened using OMIM,GeneCards,DrugBank,PharmGKB databases to obtain drug disease intersection targets.Cytoscape 3.10.1 software was used to construct the drug-component-target network and screen the key components;STRING database was used to construct protein-protein interaction(PPI)network and screen core targets;GO and KEGG enrichment analysis was conducted using DAVID platform;AutoDock Vina software was used to conduct molecular docking on the core targets and key components with high degree value respectively.CVA rat model was established by smoking combined with intraperitoneal and subcutaneous multi-point injection of ovalbumin.The rats were randomly divided into blank group,model group,montelukast sodium tablets group,Guben Zhike Pingchuan Granules low-and high dosage groups.The groups received corresponding medicine for 15 d.HE staining was used to observe the morphology of lung tissue;ELISA was used to detect the contents of serum interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α);immunohistochemistry was used to detect the positive expressions of AKT,EGFR and NFKB1 in lung tissue.Results A total of 9996 active components and 1119 potential targets of Guben Zhike Pingchuan Granules,2740 CVA targets,467 drug and disease intersection targets were screened out;PPI network topology analysis showed that AKT1,STAT3,EGFR,NFKB1 and MTOR were the core targets;the results of enrichment analysis showed that Guben Zhike Pingchuan Granules played an anti-inflammatory role in the treatment of CVA mainly through the interaction of EGFR/PI3K-Akt/NF-κB signaling pathway;molecular docking showed good binding ability between the core targets and key components,with a binding energy of<-7.0 kcal/mol for EGFR and Quercetin.Animal experiment results showed that after intervention with Guben Zhike Pingchuan Granules,the alveolar walls of the lung tissue in the model rats narrowed,the size of the alveoli became uniform,lymphocyte infiltration improved significantly,bronchioles contracted,and mucus in the lumen decreased;compared with the model group,the serum contents of IL-1βand TNF-αdecreased in Guben Zhike Pingchuan Granules low-and high dosage groups and montelukast sodium tablets group,and the positive expressions of AKT,EGFR and NFKB1 in lung tissue decreased(P<0.05,P<0.01).Conclusion Guben Zhike Pingchuan Granules may mediate the inflammatory response through the EGFR/PI3K-Akt/NF-κB signaling pathway,significantly improve the structural damage of CVA lung tissue,inhibit inflammatory cell infiltration,down-regulate the expressions of inflammatory factors,and thus exert therapeutic effects on CVA.
作者
宫雅乔
华醒实
孟莉
房小楠
甘雨
GONG Yaqiao;HUA Xingshi;MENG Li;FANG Xiaonan;GAN Yu(School of Traditional Chinese Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110032,China;The Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China)
出处
《中国中医药信息杂志》
2025年第9期31-39,共9页
Chinese Journal of Information on Traditional Chinese Medicine
基金
辽宁省科技计划联合基金项目(2023-MSLH-155)
辽宁省教育厅面上项目(LJKMZ20221330)。
