蒙药槟榔十三味丸(高尤-13)止痛作用网络药理学机制研究

Mechanism of Mongolian Binglang Shisanwei Pill on Analgesia Based on Network Pharmacology

目的:应用网络药理学、体外实验技术,预测以及验证蒙药槟榔十三味丸的止痛作用机制。方法:TCMSP数据库及文献挖掘获取槟榔十三味丸的化学成分,SwissTargetPrediction及Uniprot数据库预测成分相关靶点信息,检索OMIM等疾病靶点数据库,收集疼痛、炎症疼痛、抑郁疼痛及神经疼痛的靶点,映射至槟榔十三味丸活性成分靶点基因,构建“活性成分-靶点”网络,STRING数据库获取蛋白相互作用关系网络信息,DAVID数据库进行GO生物过程富集分析和KEGG信号通路富集分析。HEK293细胞转染TRPV1、TRPA1质粒,制备hTRPV1/HEK293细胞和hTRPA1/HEK293细胞模型,采用全细胞膜片钳技术检测槟榔十三味丸水提取物干预转染细胞后对细胞跨膜电流的影响。结果:从槟榔十三味丸的13味药材中共得86个化学成分,筛选出805个复方治疗疼痛靶点、732个治疗炎症疼痛靶点、713个治疗抑郁疼痛靶点及793个治疗神经疼痛靶点,与热敏通道(TRPA1、TRPV1及TRPV2等)相关化合物21个,共涉及135条信号通路。体外实验结果表明,槟榔十三味丸水提取物能够激活TRPA1/HEK293和TRPV1/HEK293细胞诱导其产生明显的跨膜电流。结论:槟榔十三味丸可能通过作用于TRPA1、TRPV1等热敏靶点,从而调节神经组织中配体受体的交互刺激、炎性介质对色氨酸通道及cGMP-PKG等信号通路发挥止痛作用。

Objective:To explore the underlying analgesic mechanism of Mongolian Binglang Shisanwei pill(BSP)based on network pharmacology and in vitro experimental techniques.Methods:The chemical components of 13herbs in BSP were obtained from TCMSP database,and the targets of these chemical components were predicted by SwissTargetPrediction and Uniprot database.The targets of pain,inflammatory pain,depression pain and nerve pain were collected by retrieving OMIM database,and mapped to the target genes of the active components of BSP to construct the active component-target network.The protein-protein interaction(PPI)network with common targets was constructed by using the STRING.The common target genes were enriched in GO function and KEGG pathway through DAVID database.HEK293cells were transfected with TRPV1and TRPA1plasmids to prepare hTRPV1/HEK293cells and hTRPA1/HEK293cell models.Whole cell voltage clamp technique was used to record transmembra water extracts of BSP in HEK293cells which expressed human TRPV1(hTRPV1/HEK293)and TRPA1(hTRPA1/HEK293).Results:Among the 13herbs of BSP,86chemical components,805targets for pain,732for inflammatory pain,713for depression pain,and 793for nerve pain were screened out.21compounds related to thermosensitive targets(TRPA1,TRPV1and TRPV2,etc),involving 135signal pathways.The results of in vitro experiments showed that TRPV1/TRPA1agonist was found in water extracts of BSP.Conclusion:BSP can act on TRPA1,TRPV1and other thermosensitive targets through nine Mongolian herbs such as Semen Arecae,Fructus Piperis Longi,and Semen Myristicae,thereby regulating neuroactive ligand-receptor interaction,inflammatory mediator regulation of tryptophan channels,and cGMP-PKG signaling pathways for analgesia.