[Objective]To explore the anti-cerebral ischemia mechanism of Rhubarb root and rhizome by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)combined with network pharmacology and in vitro experiment.[Methods]The active components of Rhubarb root and rhizome in rat serum samples were identified by UPLC-Q-TOF/MS.The potential targets of Rhubarb root and rhizome in blood components were obtained by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Swiss Target Prediction.The related targets of ischemic stroke(IS)were obtained from Genome Annotation Database Platform(GeneCards),Online Mendelian Inheritance in Man(OMIM)database and Therapeutic Target Database(TTD).The intersection targets were obtained by using Venny 2.1.0 software and imported into the STRING database for protein-protein interaction(PPI)network analysis.Cytoscape 3.7.2 software was employed to visualize and identify the core targets.Gene ontology(GO)enrichment function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway enrichment analysis were conducted to predict the mechanism of action.Molecular docking was performed by AutoDock Vina.Finally,the network pharmacological prediction results were verified by in vitro experiments.[Results]A total of 15 active components were identified in serum containing Rhubarb root and rhizome(SR),and there were 87 intersection targets between Rhubarb root and rhizome and IS.KEGG enrichment analysis showed that Rhubarb root and rhizome may alleviate IS through phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)and nuclear factor-κB(NF-κB)signaling pathways.Molecular docking results showed that the core components had a strong binding ability to the key targets of Toll-like receptor 4(TLR4)/myeloid differentiation primary response gene 88(MyD88)/NF-κB signaling pathway.In vitro experiment demonstrated that SR reduced the production of reactive oxygen species(ROS)induced by oxygen-glucose deprivation/reoxygenation(OGD/R)in BV2 cells.It also increased the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),while decreasing the levels of malondialdehyde(MDA)and nitric oxide(NO).Additionally,SR reduced the levels of pro-inflammatory factors,increased the levels of anti-inflammatory factors in BV2 cells induced by OGD/R,and decreased the protein expression of TLR4,MyD88 and NF-κB in BV2 cells.[Conclusion]Rhubarb root and rhizome protects BV2 microglia from OGD/R-induced injury by inhibiting oxidative stress and inflammatory response,and its mechanism may be related to regulation of TLR4/MyD88/NF-κB signaling pathway.

[Objective]To explore the anti-cerebral ischemia mechanism of Rhubarb root and rhizome by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)combined with network pharmacology and in vitro experiment.[Methods]The active components of Rhubarb root and rhizome in rat serum samples were identified by UPLC-Q-TOF/MS.The potential targets of Rhubarb root and rhizome in blood components were obtained by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Swiss Target Prediction.The related targets of ischemic stroke(IS)were obtained from Genome Annotation Database Platform(GeneCards),Online Mendelian Inheritance in Man(OMIM)database and Therapeutic Target Database(TTD).The intersection targets were obtained by using Venny 2.1.0 software and imported into the STRING database for protein-protein interaction(PPI)network analysis.Cytoscape 3.7.2 software was employed to visualize and identify the core targets.Gene ontology(GO)enrichment function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)signal pathway enrichment analysis were conducted to predict the mechanism of action.Molecular docking was performed by AutoDock Vina.Finally,the network pharmacological prediction results were verified by in vitro experiments.[Results]A total of 15 active components were identified in serum containing Rhubarb root and rhizome(SR),and there were 87 intersection targets between Rhubarb root and rhizome and IS.KEGG enrichment analysis showed that Rhubarb root and rhizome may alleviate IS through phosphatidylinositol 3-kinase/protein kinase B(PI3K/AKT)and nuclear factor-κB(NF-κB)signaling pathways.Molecular docking results showed that the core components had a strong binding ability to the key targets of Toll-like receptor 4(TLR4)/myeloid differentiation primary response gene 88(MyD88)/NF-κB signaling pathway.In vitro experiment demonstrated that SR reduced the production of reactive oxygen species(ROS)induced by oxygen-glucose deprivation/reoxygenation(OGD/R)in BV2 cells.It also increased the activity of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),while decreasing the levels of malondialdehyde(MDA)and nitric oxide(NO).Additionally,SR reduced the levels of pro-inflammatory factors,increased the levels of anti-inflammatory factors in BV2 cells induced by OGD/R,and decreased the protein expression of TLR4,MyD88 and NF-κB in BV2 cells.[Conclusion]Rhubarb root and rhizome protects BV2 microglia from OGD/R-induced injury by inhibiting oxidative stress and inflammatory response,and its mechanism may be related to regulation of TLR4/MyD88/NF-κB signaling pathway.