知柏地黄汤抗HSV-2的作用机制

The Mechanism of Zhibai Dihuang Decoction Against HSV-2

目的运用网络药理学和蛋白质组学方法探讨知柏地黄汤抗HSV-2的作用机制。方法利用中药系统药理学数据库与分析平台(TCMSP)获取知柏地黄汤的活性成分及其作用靶点。通过GeneCards和OMIM数据库筛选HSV-2的相关靶点,通过韦恩图获取二者交集靶点,构建交集靶点PPI网络并利用Cytoscape3.9.1进行可视化分析,通过DAVID数据库对交集靶点进行GO及KEGG分析,对主要有效成分与关键靶点进行分子对接。构建HSV-2感染神经元细胞(SH-SY5Y)的细胞模型,用知柏地黄汤含药血清干预HSV-2感染的人神经母细胞瘤细胞,提取细胞蛋白,运用Label free非标记定量蛋白组学技术检测蛋白表达差异,并对这些差异蛋白进行GO及KEGG富集分析。结果网络药理学共筛选出69个活性成分,213个药物靶点,282个HSV-2的靶点,37个药物靶点与疾病靶点的交集靶点。知柏地黄汤核心成分为槲皮素、山奈酚、豆甾醇等,关键靶点为IL-6、IL-10、TNF等,GO及KEGG分析结果显示主要参与炎症反应等生物过程。分子对接结果显示,主要有效成分和靶点之间具有较好的结合性。蛋白组学结果显示共有1188个蛋白参与了知柏地黄汤干预神经细胞内HSV-2的复制,显著性差异蛋白KEGG通路富集显示知柏地黄汤所涉及的通路主要有Toll信号通路、IL-17信号通路等。结论通过网络药理学和蛋白质组学研究显示知柏地黄汤主要通过白介素、Toll样受体、TNF等信号通路发挥抗HSV-2作用,通过多成分、多靶点、多通路协同发挥抗HSV-2的作用。

Objective To explore the anti-HSV-2 mechanism of Zhibai Dihuang decoction by network pharmacology and proteomics.Methods The active components and action targets of Zhibai Dihuang decoction were obtained using a traditional Chinese medicine system pharmacology database and analysis platform(TCMSP).The related targets of HSV-2 were screened by GeneCards and OMIM database,the intersection targets were obtained by Venn diagram,the PPI network of intersection targets was constructed and visualized by Cytoscape3.9.1,the intersection targets were analyzed by GO and KEGG through DAVID database,and the main effective components were docked with key targets.The cell model of neuronal cells(SH-SY5Y)infected by HSV-2 was constructed,and the human neuroblastoma cells infected by HSV-2 were treated with the serum containing Zhibai Dihuang decoction.The cellular proteins were extracted,the protein expression differences were detected by label-free quantitative proteomics technique,and GO and KEGG concentration analyses were performed for these differentially expressed proteins.Results A total of 69 active components,213 drug targets,282 HSV-2 targets and 37 intersection targets between drug targets and disease targets were screened by network pharmacology.The core components of Zhibai Dihuang decoction were quercetin,kaempferol and stigmasterol,and the key targets were IL-6,IL-10,and TNF,mainly involved in biological processes such as inflammation as found by GO and KEGG analyses.The results of molecular docking showed a good binding between the main active components and the target.Proteomic results showed that a total of 1188 proteins were involved in the intervention of Zhibai Dihuang decoction in the replication of HSV-2 in nerve cells.The enrichment of the KEGG pathway showed that the main pathways involved in Zhibai Dihuang decoction were Toll and IL-17 signaling pathways.Conclusion Employing network pharmacology and proteomics,we found that Zhibai Dihuang decoction can exert its anti-HSV-2 effect through multiple components and pathways,primary involving interleukin,Toll-like receptor,TNF and other signaling pathways.