Integrated gut microbiota,plasma metabolomics and network pharmacology to reveal the anti-ischemic stroke effect of cooked rhubarb

Background:Cooked rhubarb(CR)is obtained by steaming raw rhubarb(Rheum palmatum L.,Rheum officinale Baill.,or Rheum tanguticum Maxim.ex Balf.)with millet wine.It is used as a traditional Chinese medicine for treating cerebral stroke,where patients often face severe constipation.This study explored the pharmacological effects and mechanisms of CR against ischemic stroke(IS)in rats.We used integrated analysis of gut microbiota,metabolomics,and network pharmacology.Methods:The compounds in CR were identified using LC-MS/MS.The impact of CR on rat middle cerebral artery occlusion/reperfusion(MCAO/R)-induced cerebral infarct size and brain tissue pathology was assessed through 2,3,5-triphenyl tetrazolium chloride and HE staining.Changes in hemorheology were measured by evaluating blood viscosity,and serum levels of pro-inflammatory cytokine were also determined.Gut microbiota composition was analyzed through 16S rRNA gene sequencing.Serum metabolites were examined using untargeted metabolomics and Spearman correlation analysis.The pseudo-germ-free test was used to determine whether tumour necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)improvement in IS are dependent on gut microbiota.Results:In vivo studies showed CR enhances neurobehavioral function in MCAO/R rats and reduces cerebral infarction area.CR also ameliorates brain and intestinal barrier damage caused by stroke.It also decreased inflammation and oxidative stress in IS rats.Furthermore,CR promotes the growth of Akkermansia and Verrucomicrobia,aiding in intestinal barrier repair.Notably,CR primarily influences the primary bile acid biosynthesis pathway.The pseudo-germ-free experiment and network pharmacology confirmed TNF-αand IL-1βas potential targets.CR relies on gut microbiota for its anti-inflammatory effects to improve IS.Conclusion:Cooked rhubarb offers neuroprotective benefits by enhancing beneficial bacteria abundance and regulating bile acid metabolism.It emerges as a potential therapeutic agent for IS.

Exploration of action mechanisms of Resina Draconis(Daemonorops draco Bl.)against pain by network pharmacology and molecular docking analysis

Background:Resina Draconis is a traditional Chinese medicine mainly used to treat pain.However,the pharmacological mechanisms and chemical composition of Resina Draconis are not clear yet.Methods:In this study,based on the 21 main active components of Resina Draconis previously analyzed by our group,the potential action targets of the active components were predicted and screened out by using the databases such as Swiss Target Prediction and Pharmapper.The genes corresponding to the related targets were retrieved by UniProt and GeneCards,and then the"component-target"network model was established using Cytoscape 3.9.1 software.The protein-protein interaction network was constructed by using STRING database for analysis.The STRING database was used for enrichment analysis of gene ontology and Kyoto Encyclopedia of Genes and Genome pathways to explore the underlying action mechanisms.Result:A total of 21 main analgesic active components of Resina Draconis and 77 intersecting targets of Resina Draconis and pain were screened out.PPI network analysis indicated that such targets as albumin(ALB),tumor necrosis factor(TNF),RAC-alpha serine/threonine-protein kinase 1(AKT1)and epidermal growth factor receptor(EGFR)might be the core targets of analgesia.Through gene ontology enrichment analysis,a total of 169 gene ontology entries were obtained(P<0.01),including 111 biological processes,31 molecular functions and 27 cellular components.Through enrichment analysis of KEGG pathways,a total of 112(P<0.01)signaling pathways were screened.Conclusion:Dracaenogenins A,Resveratrol and 7,4′-dihydroxyflavone in Resina Draconis may be the main material basis for analgesia,which can interact with multiple targets such as AlB,AKT1,TNF,and EGFR,and exerts analgesic effect through signaling pathways such as mitogen-activated protein kinase(MAPK)signaling pathway,PI3K-Akt signaling pathway,and Rap1 signaling pathway.