Based On Network Pharmacology And Bioinformatics To Explore The Molecular Mechanism Of Naoshuming Prescription Against Cerebral Ischemia Injury

Objective:To explore the molecular mechanism of Nao Shuming decoction treat cerebral ischemic injury by network pharmacology and bioinformatics,then verify by animal experiments.Methods:1.First,download the original data of the disease gene chip through the Gene Expression Database(GEO)and Drugbank.Second,Traditional Chinese medicine systems pharmacology(TCMSP)was used to search for the active ingredients and targets of Nao Shuming decoction.The active ingredient-target network and protein interaction network diagrams were constructed with the String database,Perl software and Cytoscape software.Finally,gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)signaling pathway enrichment analysis were performed.It was found that Nao Shuming decoction might treat cerebral ischemia by inhibiting inflammatory reaction.2.The middle cerebral artery occlusion(MCAO)was used to duplicate the model of cerebral ischemia in rats.The rats were randomly divided into model group,normal group,sham operation group,low dose group,middle dose group and high dose group of Nao Shuming decoction.Each group was killed on the 3rd.7th and 14th day after the establishment of the model.The neurological damage was scored according to the longa 5points scoring method.The protein expression of NF-?B/p50?NF-?B/p65?TNF-? and IL-1? was detected by immunohistochemistry.The mRNA expression of NF-?B,TNF-? and IL-1? was detected by quantitative real time PCR(qRT-PCR).Results:1.It was initially found that the active components of Nao Shuming decoction are?-sitosterol,ginsenoside rh2 and naringin,the targets of Nao Shuming decoction may be PTGS1,CHRM1,SLC6A2,CHRM2,SLC6A4,BCL2,CASP3,CASP8,TNFSF15,NFKBIA,IL1B,ADCYAP1,NOS2,RELA,AKT1,MAPK3,MAPK1,BAD,GSR and other genes.2.It was found that Nao Shuming Decoction may be related to apoptosis pathway,TNF pathway,cAMP pathway,IL-17 pathway,AGE-RAGE pathway,Toll-like receptor signaling pathway and Neurotrophin signaling pathway.3.Compared with the sham group,at each time point,all of the MACO model had neurological dysfunction.But the neurological function scores will gradually improve over time.Compared with the model group,the neurological dysfunction in the high,middle and low groups of Nao Shuming decoction reduced at 3d,7d and 14d,the difference was statistically significant(P<0.05 or 0.01).4.Compared with the sham operation group,at each time point,the inflammation indexes of MACO model rats were significantly enhanced.But the inflammatory indexes will gradually improve over time.Compared with the model group,the neurological dysfunction,protein and mRNA expression levels of NF-?B?TNF-? and IL-1?3 in the high,middle and low groups of Nao Shuming decoction reduced at 3d,7d and 14d,the diffeerence was statistically significant(P<0.05 or 0.01).5.Compared with the model group,the mRNA expression levels of NF-?B?TNF-a and IL-1? in the high,middle and low groups of Nao Shuming decoction reduced at 3d,7d and 14d,the difference was statistically significant(P<0.05 or 0.01).Conclusions:1.Nao Shuming decoction has the characteristics of multi-component and multi-target synergistic effect against cerebral ischemic injury,and its mechanism may be closely related to the inhibition of inflammation.2.After cerebral ischemia,the brain's inflammatory response will be significantly enhanced and neurological dysfunction occurs.Nao Shuming decoction can reduce the expression of NF-?B,TNF-? and IL-1? in cerebral ischemic rats,and improve the neurological function.