Abstract：
OBJECTIVE: To investigate the potential mechanism of treating varicocele-associated male infertility with Jujing pill using network pharmacology and molecular docking technology.
METHODS: The TCMSP and BATMAN databases were used to search for the Chinese medicine components of the Jujing pill and obtain the corresponding targets. The databases GeneCards, DISGENET, OMIM, and HPO were searched for \'varicocele\' and \'male infertility\' to identify the potential targets of varicocele-associated male infertility. Wayne diagrams were drawn using the jvenn tool to determine the intersection targets of the Chinese medicine targets and disease targets. The intersecting targets were further analyzed to identify the components and Chinese medicine corresponding to them. A Chinese medicine-active ingredient-target network map was constructed in Cytoscape 3.8.2. The protein-protein interaction (PPI) network of the intersecting targets was constructed using the STRING platform. The intersecting targets were imported into the DAVID database for GO enrichment analysis and KEGG-based pathway enrichment analysis. The KEGG database was used to select the most relevant pathway to the topic, and a KEGG pathway map was constructed using the mapper tool. The top 15 pathways with FDR values and their related targets and components were used to draw a core ingredient-target-pathway map. Finally, molecular docking was performed to verify the protein receptors and small molecule ligands of the core genes, and the results were visualized using AutoDock and PyMol software.
RESULTS: A total of 207 ingredients and 1103 predicted targets of Jujing pill were screened. Additionally, 285 targets of varicocele were also identified. By using a Venn diagram, 86 common targets were obtained. The analysis of Gene Ontology (GO) results revealed significant enrichment in various biological processes (BP) such as positive regulation of gene expression, positive regulation of transcription, positive and negative regulation of apoptotic processes, response to hypoxia, response to estradiol, and positive regulation of nitric oxide biosynthesis processes. Furthermore, significant enrichment in cellular components (CC) was observed in macromolecules, cytoplasm, nucleus, and phosphatidylinositol 3-kinase complex. In terms of molecular function (MF), enrichment was found in enzyme binding, identical protein binding, transcriptional co-activator binding, and others. KEGG analysis demonstrated enrichment in pathways related to cancer, AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, FoxO signaling pathway, and more. Molecular docking results indicated that the core ingredients exhibited a strong binding capacity with the key targets. Conclusion： The effective active ingredients of Jujing pill exert their therapeutic effects on varicocele-associated male infertility through multiple targets and pathways. These findings provide a theoretical basis for future cell and animal experiments to verify the mechanism of action of Jujing pill in treating varicocele-associated male infertility.
METHODS: The TCMSP and BATMAN databases were used to search for the Chinese medicine components of the Jujing pill and obtain the corresponding targets. The databases GeneCards, DISGENET, OMIM, and HPO were searched for \'varicocele\' and \'male infertility\' to identify the potential targets of varicocele-associated male infertility. Wayne diagrams were drawn using the jvenn tool to determine the intersection targets of the Chinese medicine targets and disease targets. The intersecting targets were further analyzed to identify the components and Chinese medicine corresponding to them. A Chinese medicine-active ingredient-target network map was constructed in Cytoscape 3.8.2. The protein-protein interaction (PPI) network of the intersecting targets was constructed using the STRING platform. The intersecting targets were imported into the DAVID database for GO enrichment analysis and KEGG-based pathway enrichment analysis. The KEGG database was used to select the most relevant pathway to the topic, and a KEGG pathway map was constructed using the mapper tool. The top 15 pathways with FDR values and their related targets and components were used to draw a core ingredient-target-pathway map. Finally, molecular docking was performed to verify the protein receptors and small molecule ligands of the core genes, and the results were visualized using AutoDock and PyMol software.
RESULTS: A total of 207 ingredients and 1103 predicted targets of Jujing pill were screened. Additionally, 285 targets of varicocele were also identified. By using a Venn diagram, 86 common targets were obtained. The analysis of Gene Ontology (GO) results revealed significant enrichment in various biological processes (BP) such as positive regulation of gene expression, positive regulation of transcription, positive and negative regulation of apoptotic processes, response to hypoxia, response to estradiol, and positive regulation of nitric oxide biosynthesis processes. Furthermore, significant enrichment in cellular components (CC) was observed in macromolecules, cytoplasm, nucleus, and phosphatidylinositol 3-kinase complex. In terms of molecular function (MF), enrichment was found in enzyme binding, identical protein binding, transcriptional co-activator binding, and others. KEGG analysis demonstrated enrichment in pathways related to cancer, AGE-RAGE signaling pathway in diabetic complications, HIF-1 signaling pathway, FoxO signaling pathway, and more. Molecular docking results indicated that the core ingredients exhibited a strong binding capacity with the key targets. Conclusion： The effective active ingredients of Jujing pill exert their therapeutic effects on varicocele-associated male infertility through multiple targets and pathways. These findings provide a theoretical basis for future cell and animal experiments to verify the mechanism of action of Jujing pill in treating varicocele-associated male infertility.
摘要：
目的：探讨聚精丸应用网络药理学和分子对接技术治疗精索静脉曲张相关男性不育症的可能机制。
方法：利用TCMSP和BATMAN数据库检索聚精丸的中药成分并获得相应的靶点。GeneCards数据库,DISGENET,OMIM,和HPO搜索“精索静脉曲张”和“男性不育症”，以确定精索静脉曲张相关男性不育症的潜在目标。使用jvenn工具绘制Wayne图以确定中药靶标和疾病靶标的交叉靶标。对相交的靶标进行了进一步分析，以确定与之相对应的成分和中药。在Cytoscape3.8.2中构建了中药-有效成分-靶标网络图。使用STRING平台构建了相交靶标的蛋白质-蛋白质相互作用（PPI）网络。将相交的靶标导入到DAVID数据库中进行GO富集分析和基于KEGG的途径富集分析。KEGG数据库用于选择与主题最相关的途径，并使用映射器工具构建了KEGG路径图。使用具有FDR值的前15个途径及其相关靶标和组分来绘制核心成分-靶标-途径图。最后,进行分子对接以验证核心基因的蛋白质受体和小分子配体，并使用AutoDock和PyMol软件对结果进行可视化。
结果：共筛选出207个成分和1103个预测目标的聚精丸。此外,还确定了285个精索静脉曲张目标。通过使用维恩图，共获得86个目标。对基因本体论(GO)结果的分析揭示了在各种生物过程(BP)中的显著富集,如基因表达的正调控,转录的正向调节，凋亡过程的正向和负向调节，对缺氧的反应，对雌二醇的反应，和一氧化氮生物合成过程的正向调节。此外，在大分子中观察到细胞成分（CC）的显着富集，细胞质，核，和磷脂酰肌醇3-激酶复合物。就分子功能（MF）而言，在酶结合中发现了富集，相同的蛋白质结合，转录共激活因子结合，和其他人。KEGG分析显示与癌症相关的通路富集，AGE-RAGE信号通路在糖尿病并发症中的作用,HIF-1信号通路,FoxO信号通路,还有更多.分子对接结果表明，核心成分表现出与关键靶标的强结合能力。结论：聚精丸有效活性成分通过多靶点、多途径发挥对精索静脉曲张相关男性不育的治疗作用。这些发现为今后验证聚精丸治疗精索静脉曲张相关男性不育的作用机制的细胞和动物实验提供了理论依据。
方法：利用TCMSP和BATMAN数据库检索聚精丸的中药成分并获得相应的靶点。GeneCards数据库,DISGENET,OMIM,和HPO搜索“精索静脉曲张”和“男性不育症”，以确定精索静脉曲张相关男性不育症的潜在目标。使用jvenn工具绘制Wayne图以确定中药靶标和疾病靶标的交叉靶标。对相交的靶标进行了进一步分析，以确定与之相对应的成分和中药。在Cytoscape3.8.2中构建了中药-有效成分-靶标网络图。使用STRING平台构建了相交靶标的蛋白质-蛋白质相互作用（PPI）网络。将相交的靶标导入到DAVID数据库中进行GO富集分析和基于KEGG的途径富集分析。KEGG数据库用于选择与主题最相关的途径，并使用映射器工具构建了KEGG路径图。使用具有FDR值的前15个途径及其相关靶标和组分来绘制核心成分-靶标-途径图。最后,进行分子对接以验证核心基因的蛋白质受体和小分子配体，并使用AutoDock和PyMol软件对结果进行可视化。
结果：共筛选出207个成分和1103个预测目标的聚精丸。此外,还确定了285个精索静脉曲张目标。通过使用维恩图，共获得86个目标。对基因本体论(GO)结果的分析揭示了在各种生物过程(BP)中的显著富集,如基因表达的正调控,转录的正向调节，凋亡过程的正向和负向调节，对缺氧的反应，对雌二醇的反应，和一氧化氮生物合成过程的正向调节。此外，在大分子中观察到细胞成分（CC）的显着富集，细胞质，核，和磷脂酰肌醇3-激酶复合物。就分子功能（MF）而言，在酶结合中发现了富集，相同的蛋白质结合，转录共激活因子结合，和其他人。KEGG分析显示与癌症相关的通路富集，AGE-RAGE信号通路在糖尿病并发症中的作用,HIF-1信号通路,FoxO信号通路,还有更多.分子对接结果表明，核心成分表现出与关键靶标的强结合能力。结论：聚精丸有效活性成分通过多靶点、多途径发挥对精索静脉曲张相关男性不育的治疗作用。这些发现为今后验证聚精丸治疗精索静脉曲张相关男性不育的作用机制的细胞和动物实验提供了理论依据。
