Based On Network Pharmacology, Research On The Mechanism Of Action Of The Multi-active Ingredients Of Panax Notoginseng To Protect The Neurovascular Unit Of Cerebral Ischemia

Objective: The related genes and pathways of neurovascular unit(NVU)affected by ischemic stroke were analyzed;The differentially expressed genes(DEGs)of Panax notoginseng(PN)on middle cerebral artery occlusion(MCAO)rats were analyzed by RNAseq,And explore the potential mechanism of the neuroprotection of PN;Network pharmacology analysis of PN active ingredients,multi-target regulation and multi-channel protection of the key genes and pathways of NVU in ischemic stroke.Methods:(1)Through the GEO database mining,we integrate the GEO gene expression chip of the main components of NVU(neurons,astrocytes,vascular endothelial cells and peripheral cells),and analyze the key genes and pathways that affect the ischemic stroke.(2)The DEGs between the intervention group and the model control group were identified by RNA-seq.The possible mechanism of PN involved in the regulation of ischemic stroke was analyzed.(3)The main active components of PN were selected from the authoritative TCMSP database,and the targets of the active components were identified with multiple databases,and integrated with the related genes of the first two parts of the study to analyze the mechanism of PN protecting the neurovascular unit of ischemic stroke.Results:(1)The neuron GSE54037 chip screened 32 target genes of ischemic stroke,including 6 up-regulated genes and 26 down-regulated genes.The GSE3045 chip of astrocytes screened 130 target genes of ischemic stroke,90 of which were up-regulated genes and 40 of which were down-regulated genes.GSE76739 chip of vascular endothelial cells screened 69 target genes of ischemic stroke,48 of which were up-regulated genes and 21 of which were down-regulated genes.24 target genes of ischemic stroke were screened by GSE109233.236 NVU targets were obtained by integrating the target genes of 4 chips.Go analysis showed that these target genes are mainly related to biological processes such as biological adhesion,immune system process,cell proliferation,response to stimulation,metabolic process,cell killing and synapse.The signal pathways of KEGG enrichment were PI3K-Akt,HIF-1,complement coagulation cascade,cytokine receptor interaction pathway and so on.Through the topological analysis of the PPI network by the software of Cytoscape,the connectivity of VEGFA,IL6,CXCL8,and SRC in the whole network is more than 30,which is the key gene of this study.(2)Compared with the model group,the neurological deficit score of MCAO rats treated with PN powder decreased significantly(P < 0.01).RNA-seq identified 817 DEGs(390 up-regulated genes and 427 down-regulated genes)in PN/MCAO group.The GO analysis of DEGs is helpful to reveal the biological process of PN's anti-ischemic effect,including the regulation of cell cycle metabolism,vascular development,and neurotransmitter transport.Molecular functions include nucleic acid binding,ion binding,and DNA binding.Cell components mainly include synapse,postsynaptic,glutamatergic synapse and chloride channel complex.KEGG pathway analysis mainly involves PI3K-Akt signaling pathway,calcium signaling pathway,CGMP-PKG signaling pathway,AMPK signaling pathway,etc.(3)Through multi database mining,12 active components of PN were identified corresponding to 1755 targets,420 of which were involved in the regulation mechanism of PN against ischemic stroke,and 236 targets of NVU were analyzed together.48 key targets were identified,including IL6,VEGFA,and CXCL8,which were mainly involved in the regulation of NF-?B signaling pathway(CXCL8,SYK,PLAU,VCAM1,PARP1),p53 signaling pathway(CDK2,IGF1,IGFBP3,Seri1),PI3K-Akt signaling pathway(IL6,MMP2,SPP1,VEGFA,IGF1,CDK2,SYK),Toll-like receptor signaling pathway(CXCL8,IL6,SPP1).Conclusion:(1)The key genes(VEGFA,IL6,CXCL8,SRC)and regulatory pathways(PI3K-Akt signaling pathway,HIF-1 signaling pathway,complement coagulation cascade signaling pathway)that may be involved in the mechanism of ischemic stroke were obtained through the integrated analysis of gene chip of NVU.(2)PN powder can improve the symptoms of neurological deficits in rats with ischemic stroke.The potential neuroprotective mechanism may be to regulate PI3K-Akt signal pathway,calcium signal pathway,CGMP-PKG signaling pathway and AMPK signal pathway to play a role.(3)PN has multiple active components,which act on NVU network through multiple targets and pathways.IL6,VEGFA,CXCL8 are potential key genes,and quercetin may play an important role in the pharmacological action of Panax notoginseng.