基于网络药理学联合分子对接探析强精片治疗男性不育症的分子机制

Exploring the Molecular Mechanism of Qiangjing Tablets in the Treatment of Male Infertility Based on Network Pharmacology and Molecular Docking

目的通过网络药理学及分子对接技术探析强精片治疗男性不育症的有效成分和潜在分子机制,并对不育症模型小鼠进行相关作用靶点的实验验证。方法使用中药系统药理学分析平台(TCMSP)收集强精片的活性成分和潜在靶点,利用Genecard数据库筛选男性不育症的疾病靶点;然后将疾病靶点与药物预测靶点取交集,借助Cytoscape3.7.2软件构建交集靶点网络以及蛋白质-蛋白质相互作用网络,利用Bioconductor平台和R语言进行GO和KEGG富集分析。采用氯化镉注射造模不育症小鼠模型,采用HE染色、电镜观察睾丸结构,qPCR检测Pi3kcb、Akt1表达量,探讨强精片治疗不育症的潜在机制。并将药物活性成分与疾病靶点进行分子对接,探讨潜在作用成分及靶点机制。结果共获得109个强精片活性成分及286个可用于后续分析的靶点,476个男性不育症靶点,映射得出69个药物-疾病共同靶点。GO分析提示主要与营养、抗氧化、对类固醇激素反应等生物过程及功能相关。KEGG通路分析提示主要与PI3K-Akt、TNF、内分泌抵抗等信号通路相关,PI3K-Akt是核心信号通路。强精片可以改善不育症模型小鼠睾丸结构,提高Pi3kcb、Akt1 mRNA表达。强精片中活性成分槲皮素、木犀草素、山奈酚、β-谷固醇、花生四烯酸与靶标蛋白MAPK1、AKT1、HSP90AA1、IL6、ESR1结合稳定。结论本研究初步明确了强精片治疗男性不育症的主要靶标和通路,为后续深入研究提供一定参考。

Objective To explore the effective ingredients and potential molecular mechanisms of Qiangjing Tablets in the treatment of male infertility through network pharmacology and molecular docking technology,and to carry out experimental verification of the relevant targets of infertility model mice.Methods We used the TCMSP to collect the active ingredients and potential targets of Qiangjing Tablets,used the Gene Card database to screen the disease targets of male infertility,then took the intersection of the disease targets and the drug prediction targets.We integrated the intersection target network and protein-protein interaction network through Cytoscape 3.7.2 software.We used the Bioconductor platform and R language for GO and KEGG enrichment analysis.A mouse model of infertility was modeled by cadmium chloride injection.The testicular structure was observed by HE staining and electron microscope,and the expression of Pi3kcb and Akt1 was detected by qPCR to explore the potential mechanism of Qiangjing Tablets in the treatment of infertility.The active ingredients of the drug were molecularly docked with the disease target.Results A total of 109 active ingredients of Qiangjing Tablets and 286 targets for subsequent analysis,476 targets for male infertility were obtained,and 69 common drug-disease targets were mapped out.GO analysis suggested that it was mainly related to biological processes and functions such as nutrition,anti-oxidation,and response to steroid hormones.KEGG pathway analysis suggested that it was mainly related to PI3K-Akt,TNF,endocrine resistance and other signaling pathways.PI3K-Akt was the core signaling pathway.Qiangjing Tablets improved the testis structure of infertility model mice,and increased the expression of Pi3kcb and Akt1 mRNA.The active ingredients in Qiangjing Tablets were quercetin,luteolin,kaempferol,β-sitosterol,arachidonic acid and the target protein MAPK1,AKT1,HSP90AA1,IL6,and ESR1 were stable in combination.Conclusion This study preliminarily clarifies the main targets and pathways of Qiangjing Tablets in the treatment of male infertility,and provided a certain reference for follow-up in-depth research.