In Vivo Validation Of Resveratrol Alleviating Intervertebral Disc Degeneration And Prediction Of Potential Targets And Mechanisms Based On Network Pharmacology

Objective: In this article,the effects of resveratrol on intervertebral disc degeneration were evaluated through the rat intervertebral disc degeneration model,and the potential targets and pathway mechanisms of resveratrol regulation of intervertebral disc degeneration were explored through network pharmacology,which laid a foundation for subsequent studies for the application of resveratrol on intervertebral disc degeneration.Methods: A rat model of intervertebral disc degeneration was established,and the degenerative segments were evaluated by X-ray and MRI after 4 and 8 weeks.Safranin O-fast green staining was used to evaluate the intervertebral disc tissue after 8 weeks.Disease-related targets of intervertebral disc degeneration was obtained and unified using Genecards,Dis Ge NET,OMIM databases.Swiss Target Prediction,TCMSP and TTD databases were used to obtain the drug targets of resveratrol,disease-related drug targets were then established.The common targets of resveratrol and intervertebral disc degeneration were obtained through the SRplot online platform and Venn diagrams were drawn.Common targets were imported into S TRING database to establish contact and eliminate free targets to construct protein interaction network.C ytoscape3.7.2 analysis was used for optimization,and the "Analyze Network" tool was used for topological analysis.The degree,betweenness centrality and closeness centrality were used as selection criteria to screen the key targets in the process of resveratrol treatment of intervertebral disc degeneration.The Metascape platform was used for G ene Ontology function and Kyoto Encyclopedia of Genes and Genomes pathways enrichment analysis,and the SRplot online platform was used for visualization of enriched pathways.3D structure of the protein was obtained from PDB database.Auto Dock Tools 1.5.7 was used for dehydration,hydrogenation,and computing charges.Then Autodock 4.2 was used to dock the ligands and receptors.The docking models of the lowest docking energy were visualized with Py MOL 2.5.0 and BIOVIA Discovery Studio 2020.Results: In vivo experiments showed that intervertebral disc degeneratio n could lead to the narrowing of intervertebral space height,the reduction of T2 WI signal intensity of nucleus pulposus,the reduction of nucleus pulposus and extracellular matrix,and the disorder of annulus fibrosus structure.Resveratrol treatment could effectively delay the changes caused by intervertebral disc degeneration.Network pharmacology was used to establish the resveratrol target library and the intervertebral disc degeneration target library.A total of 1172 targets for intervertebral disc degeneration and 203 targets for resveratrol were obtained,and 81 targets were common to both.Network pharmacological analysis was performed to 81 common targets,and the intervertebral disc degeneration-related targets of resveratrol included SRC,MAPK3,MAPK1,TP53,FOS,EGFRAKT1,STAT1,ESR1,C TNNB1,HIF1 A,PIK3CA,MAPK8,VEGFA,MYC,IL6,TGFB1,EDN1 and CDKN 1A,PPARG,CASP3,TNF,FGF2,MMP2,PRKCA,MTOR,BRCA1,etc.Molecular docking suggested that resveratrol had a high binding affinity with TP53 and MAPK3.Combination of the information of core targets and the results of enrichment analysis of cross targets suggests that PI3K-Akt,MAPK and p53 pathways are important pathways related to the mechanism of resveratrol alleviating intervertebral disc degeneration.Conclusion: Resveratrol can alleviate the progression of intervertebral disc degeneration possibly by activating PI3K-Akt pathway and inhibiting MAPK and p53 pathways.