The bio-active components of the Mongolian medicine Horcha-6 and therapeutic mechanism in the rat migraine model

Background:The active components of Horcha-6 were identified using liquid chromatography with tandem mass spectrometry.Also,we investigated the potential mechanisms that explain why Horcha-6 may be effective in treating migraines through the use of network pharmacology and a rat migraine model.Methods:After identifying the active components of Horcha-6,the corresponding genes of the active components’target were obtained from the Universal Protein database,and a“compound-target-disease”network was constructed using Cytoscape 3.9.0 software.For the in vivo experiments,nitroglycerin was injected intraperitoneally into rats to create a migraine model.Pre-treatment with Horcha-6 was administered orally for 14 days,and rats were subjected to migraine-related behavior tests.RNA sequencing was performed to identify the gene expression regulated by Horcha-6 in the trigeminal nerve.Results:A total of 903 chemical components of Horcha-6 have been collected in the liquid chromatography with tandem mass spectrometry.We discovered 55 of the Horcha-6 bio-active components that were evaluated based on their Percent Human Oral Absorption(≥30%)and DL values(≥0.185)on the traditional Chinese medicine systems pharmacology database.The“compound-target-disease”network contained 163 intersection targets with the migraine state.Gene Ontology analysis indicated that these components significantly regulated the immune response,vascular function,oxidative stress,etc.When Kyoto Encyclopedia of Genes and Genomes enrichment analysis was performed,we observed that most of the target genes were significantly enriched in the inflammation and neuro-related signaling pathway,toll-like receptor signaling pathway,neuroactive ligand-receptor interaction,etc.These predictions were further demonstrated via in vivo animal model experiments.The RNA sequencing results showed that 41 genes were down-regulated(P<0.05)and 86 genes were up-regulated(P<0.05)in the Horcha-6 treated group compared with the untreated group.Those genes were mainly involved in neuromodulation,vascular function,and hormone metabolism.Conclusion:The 55 bio-active components in Horcha-6 regulate inflammation,hormone metabolism,and neurotransmitters and have potential as a therapy to treat migraines.

Study on the Anti-inflammatory Mechanism of Hei-Yun-Xiang (黑云香) by Network Pharmacology and Molecular Docking

Objective:To identify the secondary metabolites of Hei-Yun-Xiang(黑云香),predict its target by network pharmacology,and the target binding activity was demonstrated by molecular docking to explore its mechanism in treating inflammatory diseases.Methods:The Untargeted Metabolomics was used to identify the total components of secondary metabolites in Hei-Yun-Xiang.The Hei-Yun-Xiang's active components were screened by the oral bioavailability>30%and drug-likeness>0.10 from the total component of Hei-Yun-Xiang.The TCMSP and standard chemical databases screened the main active components.The Swiss target prediction and the UniProt database predicted the potential targets of Hei-Yun-Xiang's active components.To obtain the potential targets of Hei-Yun-Xiang in the treatment of inflammatory diseases,we compare the above targets with OMIM,NCBI,and GENECARD.Combined with string analysis of Hei-Yun-Xiang's target protein interaction,the R4.0.3 software was used to enrich and analyze Hei-Yun-Xiang's potential targets'GO annotation and KEGG pathway.The"Mongolian medicine-active component-core target-pathway"network structure diagram of Hei-Yun-Xiang in anti-inflammatory effect was constructed using Cytoscape 3.8.0 software.AutoDock Vina 1.1.2 molecular docking software was used to target proteins and the active components interaction model.Results:22 highly active compounds were screened from Hei-Yun-Xiang,of which 19 were related to inflammation.324 targets related to inflammation were predicted and analyzed;targets were mainly composed of genes such as inflammatory biological processes induced by bacterial molecules such as lipopolysaccharide,cell membrane function,and serine/threonine kinase activation.Targets were enriched into the 179 KEGG-related pathways.Pathways mainly include lipid metabolism,arteriosclerosis,neuroactive receptor-ligand binding pathway,PI3K Akt signaling pathway.Molecular docking demonstrated that astrapterocarpan,chimonanthine,columbianetin_acetate,tretinoin,tussilagone,columbianetin_acetate,n-Feruloyltyramine had good binding to eight key proteins,and AKT1 and HSP90AA1 was the target protein with the best binding activity,suggesting that AKT1 and HSP90AA1 could be the essential mediator responsible for signaling transduction after Hei-Yun-Xiang administration.Conclusion:Hei-Yun-Xiang can inhibit the inflammatory reaction by its multi-component,multi-target,and multi-channel treatment of inflammatory disease,interfering with the inflammatory reaction induced by bacterial molecules such as lipopolysaccharide or by intervening in lipid and inflammation-related arteriosclerosis-related pathways.