基于网络药理学及分子对接技术探究北沙参-玉竹治疗卵巢早衰的作用机制

Exploration on the Mechanism of the Treatment of Premature Ovarian Failure by Coastal Glehnia Root-Polygonatum Odoratum Based on Network Pharmacology and Molecular Docking

目的基于网络药理学及分子对接技术,预测清肺养阴药对北沙参-玉竹改善卵巢早衰(Premature ovarian failure,POF)的关键靶点及作用通路,并通过动物实验验证有关结果。方法通过人类在线孟德尔遗传数据库(Online Mendelian Inheritance in Man,OMIM)、Drugbank、GeneCards数据库收集卵巢早衰相关靶点;通过中药系统药理学数据库与分析平台(Traditional Chinese Medicine Database and Analysis Platform,TCMSP)筛选北沙参-玉竹药对的有效活性成分,SwisstargetPredicition数据库中预测作用靶点。利用STRING平台对药对-卵巢早衰交集靶点进行蛋白质相互作用分析,根据拓扑参数筛选关键靶点。基于Metascape数据库、Cluego插件进行基因本体(Gene Ontology,GO)分析和基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)信号通路富集分析。通过Autodock vina 1.1.2软件对关键靶点和有效活性成分进行半柔性对接。使用雷公藤多苷片诱导小鼠卵巢早衰疾病模型进行实验验证。结果得到药对16个有效活性成分,协同作用于38个靶点,参与血管内皮细胞迁移、磷脂酶活性的正调控,自噬、细胞凋亡的负调控,以及肽基-丝氨酸磷酸化的调节、雌激素代谢等44个生物学过程,涉及PI3K/Akt/mTOR、Foxo、HIF-1、TNF、IL-17、MAPK、NF-κB等51条信号通路。分子对接结果显示关键靶点与有效成分间均有较强的结合能力。动物实验结果证明,北沙参-玉竹可通过上调蛋白激酶B(Akt)、磷脂酰肌醇3-激酶(PI3K),下调磷酸酶-张力蛋白基因(PTEN)、核转录因子-κB(NF-κB)的表达改善卵巢功能(P<0.01),验证了网络药理学部分预测结果。结论北沙参-玉竹可通过上调Akt、PI3K,下调PTEN、NF-κB的表达改善卵巢功能,初步证实北沙参-玉竹治疗卵巢早衰的部分作用靶点,为其临床应用及作用机制研究提供了思路和依据。

Objective Based on network pharmacology and molecular docking,the key targets and action pathways of couplet medicinals for clearing lung and nourishing yin(Coastal glehnia root-Polygonatum odoratum)in improving premature ovarian failure(POF) were predicted, and related results were verified through animal experiments.Methods The related targets of POF were collected by Online Mendelian Inheritance in Man(OMIM),Drugbank,and GeneCards database. The active components of Coastal glehnia root-Polygonatum odoratum couplet medicinals were screened by traditional Chinese medicine database and analysis platform(TCMSP), and the targets were predicted by SwisstargetPredicition database. The protein interaction analysis of herb pairs-POF intersection target was performed by using the STRING platform,and the key target was selected in accordance with the topological parameters. The Gene Ontology(GO) analysis and signal pathway enrichment analysis of Kyoto Encyclopedia of Genes and Genomes(KEGG)were carried out based on Metascape database and cluego plug-in. The semi-flexible docking of the key target to the active component was conducted by Autodock vina1.1.2 software. Mice model with tripterygium glycosides-induced POF was used to verify the simulated results.Results Sixteen active components of the herb pairs, which synergistically acting on 38 targets were obtained. They mediated 44 biological processes,including the positive regulation of vascular endothelial cell migration and phospholipase activity, the negative regulation of autophagy and apoptosis,and the regulation of peptide-serine phosphorylation and estrogen metabolism.51 signaling pathways, including PI3K/Akt/mTOR, Foxo, HIF-1, TNF, IL-17, MAPK, NF-kB etc. were involved. The results of molecular docking showed that there was strong binding capacity between the key targets and the active components. The results of animal experiments suggested that Coastal glehnia root-Polygonatum odoratum can improve ovarian function(P<0.01)by up-regulation of Akt and PI3K,down-regulation of PTEN and NF-κB,which verified partial prediction results of the network pharmacology.Conclusion Coastal glehnia root-Polygonatum odoratum can improve ovarian function by up-regulation of Akt, PI3K and down-regulation of PTEN, NF-κB.Partial targets for the action of Coastal glehnia root-Polygonatum odoratum in the treatment of POF had been preliminarily confirmed. The study provides ideas and basis for its clinical application and mechanism research.