基于网络药理学、分子对接和实验验证探讨感冒清热片抗肺损伤的作用机制  

Exploration on the Mechanism of Ganmao Qingre Pills Against Lung Injury Based on Network Pharmacology,Molecular Docking and Experimental Verification

作　　者：任薇 徐明明 高欣 马博文 苏子仁[4] 刘煜洪 赵芳宇 REN Wei;XU Mingming;GAO Xin;MA Bowen;SU Ziren;LIU Yuhong;ZHAO Fangyu(Guangdong Provincial Key Laboratory of Research and Development in Traditional Chinese Medicine,Guangdong Second Traditional Chinese Medicine Hospital(Guangdong Province Engineering Technology Research Institute of T.C.M.),Guangzhou 510095 Guangdong,China;Hebei Tangwei Pharmaceutical Co.,Ltd,Cangzhou 061000 Hebei,China;Pharmaceutical Department,Cangzhou People's Hospital,Cangzhou 061000 Hebei,China;School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China)

机构地区：[1]广东省第二中医院(广东省中医药工程技术研究院),广东省中医药研究开发重点实验室,广东广州510095 [2]河北唐威药业有限公司,河北沧州061000 [3]沧州市人民医院药学部,河北沧州061000 [4]广州中医药大学中药学院,广东广州510006

出　　处：《中药新药与临床药理》2024年第9期1376-1388,共13页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：广东省中医药局课题(20222009)。

摘　　要：目的基于网络药理学、分子对接和体内实验方法探讨感冒清热片抗肺损伤的作用机制。方法以中药系统药理学数据库和分析平台(TCM-SP)、Genecards等数据库,筛选感冒清热片治疗肺损伤的潜在作用靶点。采用Cytoscape 3.9.0软件构建“中药-有效成分-靶点”网络,并用生物信息云平台对潜在作用靶点进行基因本体(GO)功能与京都基因和基因组百科全书(KEGG)通路富集分析。建立蛋白互作(PPI)网络,并与“中药-有效成分-靶点”网络进行交集分析,筛选关键靶蛋白。将关键靶蛋白和有效成分进行分子对接。建立肺损伤小鼠模型验证感冒清热片对关键靶标蛋白的作用。结果共筛选出感冒清热片治疗肺损伤的作用靶点707个,对应11味中药中107种化合物。利用构建的“中药-有效成分-靶点”网络发现感冒清热片可能通过豆甾醇、芦丁、金合欢素等有效成分对前列腺素G/H合成酶1(PTGS1)、雄激素受体(AR)、乙酰胆碱酯酶(ACHE)等作用靶点进行调控。利用PPI网络发现SRC酪氨酸激酶(SRC)、表皮生长因子受体(EGFR)、信号转导因子和转录激活因子3(STAT3)等核心靶点。通过PPI网络与“中药-有效成分-靶点”网络交集分析,筛选出关键靶标蛋白周期蛋白依赖性激酶1(CDK1)、CDK2、EGFR、雌激素受体1(ESR1)和SRC。分子对接研究显示,感冒清热片中的豆甾醇、芦丁、金合欢素分别与CDK1、CDK2、EGFR、ESR1、SRC有较好的结合能力。体内实验发现感冒清热片可剂量依赖性地减轻肺损伤小鼠的肺组织损伤、炎性浸润,降低TNF-α、IL-1β、IL-6的含量,增强CDK1、CDK2的表达,降低EGFR、ESR1和SRC的表达。结论感冒清热片对肺损伤的治疗作用可能通过其所含的豆甾醇、芦丁、金合欢素等关键活性成分,作用于CDK1、CDK2、EGFR、ESR1、SRC等关键靶标蛋白,调控神经活性配体-受体相互作用、癌症途径、钙信号通路等关键信号通路�Objective To investigate the mechanism of Ganmao Qingre Pills(GQP)against lung injury based on network pharmacology,molecular docking and in vivo experiments.Methods The potential targets of GQP in the treatment of lung injury were screened through traditional Chinese medicine systems pharmacology database and analysis platform(TCM-SP)and Genecards.A“Chinese medicine-active ingredients-targets”network was constructed using Cytoscape 3.9.0 software,then gene ontology(GO)function and Kyoto encyclopaedia of genes and genomes(KEGG)pathway enrichment analysis for potential targets were conducted using a bioinformatics cloud platform.We established a protein-protein interaction(PPI)network,which was intersected with“Chinese medicine-active ingredients-targets”network to obtain core targets.The molecular docking between key target proteins and active ingredients was performed.The effect of GQP on these key target proteins was verified by using a mouse model of lung injury.Results A total of 707 targets for the treatment of lung injury by GQP were identified,corresponding to 107 active ingredients in 11 Chinese medicines.It was found that GQP might regulate targets such as PTGS1,AR,and ACHE through active ingredients including stigmasterol,luteolin,and acacetin using the"Chinese medicine-active ingredients-targets"network analysis.Core targets such as SRC,EGFR,and STAT3 were discovered by using the PPI network.Key target proteins,including CDK1,CDK2,EGFR,ESR1 and SRC,were screened through the intersection analysis of the PPI network and“Chinese medicine-active ingredients-targets”network.Molecular docking study showed that stigmasterol,luteolin and acacetin had good binding effects with CDK1,CDK2,EGFR,ESR1,and SRC,respectively.In vivo experiments revealed that GQP dose-dependently attenuated lung injury and inflammatory infiltration,reduced the release of pro-inflammatory factors TNF-α,IL-1βand IL-6,increased the expression of CDK1 and CDK2,and decreased the expression of EGFR,ESR1 and SRC in lung injury mi

关 键 词：感冒清热片 肺损伤 宣肺止咳 网络药理学 分子对接 实验验证 小鼠 

分 类 号：R285.5[医药卫生—中药学]