基于网络药理学和实验验证探究金银花-黄连治疗咽炎的作用机制

Mechanism Research on Lonicerae Japonicae Flos and Coptidis Rhizoma on Pharyngitis Based on Network Pharmacology and Experimental Verification

目的通过网络药理学方法对金银花-黄连治疗咽炎的作用机制进行预测,并进行初步动物实验验证。方法通过TCMSP数据库筛选金银花与黄连的有效成分;输入Pharm Mapper数据库获得相应基因;检索GeneCards数据库获得咽炎的靶点基因;利用R语言3.6.1截取交集网络获得候选靶点;借助Cytoscape3.7.1将“药物-化合物-靶点-疾病”网络可视化处理;在STRING平台构建PPI网络;利用DAVID数据库对候选靶点进行GO富集分析和KEGG通路注释;使用AutoDock Vina 4.2.6,将关键靶点与关键成分进行分子对接,并使用PyMol进行可视化处理。最后通过急性咽炎大鼠实验模型验证金银花-黄连对两种靶点基质金属蛋白酶2(MMP-2)、白介素2(IL-2)的影响。结果共获得33个有效化合物,183个不重复的靶标信息,46个疾病治疗靶点;GO与KEGG富集分析分别得到48条目和48条信号通路,主要涉及雌激素信号通路、促性腺激素释放信号通路、Ras信号通路、T细胞受体信号通路。分子对接结果显示金银花-黄连中有效成分黄柏酮、掌叶二蒽酮A与关键靶点MMP-2、表皮生长因子受体(EGFR)、丝裂原活化蛋白激酶1(MAPK1)具有自发亲和力。动物实验证实金银花-黄连高、中、低剂量能降低急性咽炎大鼠血清中的MMP-2、IL-2水平。结论通过网络药理学和实验验证,初步预测了金银花-黄连治疗咽炎可能的药效物质基础及其分子机制,可为今后的研究提供依据和方向。

Objective To explore the action mechanism of Lonicerae Japonicae Flos and Coptidis Rhizoma on pharyngitis using the method of network pharmacology and preliminary animal experiment.Methods The active ingredients of Lonicerae Japonicae Flos and Coptidis Rhizoma were screened by TCMSP database,and the corresponding genes were obtained after information input of the key compounds into Pharm Mapper database.The target genes of pharyngitis were obtained by searching GeneCards database.The candidate targets were obtained by intercepting the intersection network using R language 3.6.1,and the“drug-compound-target-disease”network was visualized using Cytoscape 3.7.1.Then PPI network was constructed on STRING platform.GO enrichment analysis and KEGG pathway annotation on candidate targets were performed using DAVID database.AutoDock Vina 4.2.6 was used for molecular docking of key targets and key components,and PyMol was used for visual processing.Finally,the effects of Lonicerae Japonicae Flos and Coptidis Rhizoma on MMP-2 and IL-2 was verified by the experimental model of acute pharyngitis in rats.Results 33 effective compounds,183 nonrepeating target information,and 46 disease treatment targets were obtained.GO and KEGG enrichment analysis yielded 48 items and 48 signaling pathways,respectively,mainly involving the estrogen signaling pathway,gonadotropin release signaling pathway,Ras signaling pathway,and T cell receptor signaling pathway.The results of molecular docking showed that obacunone and palmidin A,the active components of Lonicerae Japonicae Flos and Coptidis Rhizoma,had spontaneous affinity with key targets MMP-2,EGFR,and MAPK1.Animal experiment confirmed that the high,medium and low dose of Lonicerae Japonicae Flos and Coptidis Rhizoma could reduce the levels of MMP-2 and IL-2 in serum of rats with acute pharyngitis.Conclusion Through network pharmacology and experimental verification,the possible pharmacodynamic material basis and molecular mechanism of Lonicerae Japonicae Flos and Coptidis Rhizoma in the treatment of pharyngitis are preliminarily predicted,so as to provide basis and direction for future research.