The Protective Effect And Regulation Mechanism Of Oxyresveratrol On LPS-induced Neuroinflammation

Objective:In this article,network pharmacology was used to study the potential targets and mechanistic pathways of oxyresveratrol in regulating neuroinflammation and combined with the cell and animal models of lipopolysaccharide(LPS)-induced neuroinflammation to explore the protective effects and the relevant mechanism of oxyresveratrol on neuroinflammation.The final results will provide a more comprehensive scientific basis for the development and application of oxyresveratrol in the field of neurodegenerative diseases.Methods:1.Use network pharmacology simulation to calculate the relevant antineuroinflammation targets and mechanism of oxyresveratrol.The specific steps are as follows:Use Gene Cards,Swisstarget Prediction,Targetnet,Uniprot,Venny,String,and other network pharmacology-related databases and tools to collect neuroinflammation-related targets and oxyresveratrol regulatory targets to establish oxyresveratrol Targets library and neuroinflammation-related targets library.The Venny database is used to analyze the oxyresveratrol-neuroinflammation target intersection.The STRING database is used to establish the connection between the targets,and Cytoscape 3.7.2 is used to analyze the protein binding degree between the target and the target.The degree value,betweenness centrality value,and closeness centrality value are analyzed by the Network Analyze function in the Cytoscape software.Select the target with a score greater than the median value,and summarize it as the potential anti-neuroinflammation targets of oxyresveratrol.GO enrichment analysis and KEGG pathway annotation are used to obtain the mechanism pathway enrichment map of oxyresveratrol in regulating neuroinflammation,and analyze the possible mechanism of oxyresveratrol in regulating neuroinflammation;2.Establish LPS-induced BV2 cells to simulate in vitro neuroinflammation models.determine the safe concentration of oxyresveratrol through cytotoxicity experiments.the NO content in the cell supernatant after culturing cells with different concentrations of oxyresveratrol is measured to evaluate the effect of oxyresveratrol on NO secretion induced by LPS;3.Establish a mouse model of LPS-induced neuroinflammation to study the protective effect and regulation mechanism of oxyresveratrol on LPS-induced neuroinflammation.The mice were fed for 3 days after the adaptation period,and the administration period was 14 days.After 7 days of intraperitoneal injection of LPS for modeling,the Morris water maze experiment was carried out to record the performance of the mice in the directional navigation experiment and space exploration experiment.After the behavioral test was completed,the mice were dislocated and sacrificed.The molecular mechanism of veratrol regulating neuroinflammation was further analyzed by Nissl staining experiment,Western blotting,immunohistochemistry,enzyme-linked immunosorbent assay,real-time quantitative PCR,and tissue immunofluorescence.Results:1.Use network pharmacology simulation to establish oxyresveratrol target library(214targets)and neuroinflammation target library(1204 targets).69 common targets were obtained by cross-comparison.The network pharmacological analysis of 69 common targets was performed to obtain the anti-neuroinflammation targets of oxyresveratrol,including SRC,EGFR,ESR1,PIK3 CA,TNF,PTGS2,APP,MMP9,IGF1 R,MAP2K1,RELA,MCL1,RAC1,MAPT,PTPN1,GRIN2 B,PTK2,PRKACA,KIT,RAF1,HNF4 A,SYK.The Mechanism pathways enrichment analysis results of the 69 common targets show that the PI3K-Akt pathway and the NF-κB pathway may be the related mechanisms of oxyresveratrol in regulating neuroinflammation.2.The results of cytotoxicity experiments confirmed that the concentration range of oxyresveratrol is 200 μM and below.Oxyresveratrol at a concentration of 25 μM and above can significantly inhibit the expression of NO-induced by LPS,and oxyresveratrol inhibited the secretion of NO in a dose-dependent manner.3.In the mouse model of neuroinflammation induced by LPS,oxyresveratrol significantly improved cognition impairment and alleviated the neuronal apoptosis caused by LPS.The protective effect of oxyresveratrol on neuroinflammation is mainly through inhibiting the phosphorylation process of the PI3K/Akt/NF-κB pathway to downregulate the expression of TNF-α,IL-1β,i NOS,COX2,MMP9.The oxidative stress and the excessive activation of microglia and astrocytes was suppressed in the brain to alleviate neuroinflammation.Conclusion:Oxyresveratrol has a protective effect on LPS-induced neuroinflammation.Its regulatory mechanism is basically consistent with the predicted results of network pharmacology.Oxyresveratrol inhibited the phosphorylation of PI3K/Akt/NF-κB pathway and downregulated the expression of downstream inflammatory mediators.At the same time,Oxyresveratrol activation of the glial cells and oxidative stress induced by LPS,thereby inhibiting neuronal apoptosis and restoring cognitive impairment and memory impairment.