Efficacy And NF-?B Signaling Pathway Of Extracts From The Leaves Of Li Yao Partridge In The Treatment Of Ischemic Stroke Nerve Injury

Object:Ischemic stroke belongs to the category of "adverse wind" in traditional Li medicine,which is one of the common diseases in Hainan Province,and also a difficult problem in modern medical research.Now,ischemic stroke has been a severe problem.It has great meaningful finding potential drugs for the treatment of ischemic stroke from traditional national drug resources.The Mallotus furetianus(ZGY)is a characteristic medicine in Hainan,which has been used for hundreds of years.Modern research showed that the chemical components of ZGY has neuroprotective effect but the related pharmacological research is still insufficient,the targets are still unknown.It is urgent to use advanced technology and sufficient pharmacological research to provide more experimental data for ZGY.In this paper,we clarified the potential targets of ZGY against ischemic stroke,and established in vivo and in vitro models of ischemic stroke to study the pharmacodynamic action and molec?Lar mechanism of ZGY on ischemic stroke.Method:1.Search the genome of ischemic stroke patients and the targets of ZGY from published literature and online databases and use the intersection to obtain common target genes.With network pharmacology technology,the component-target network,protein-protein interaction network,Go and KEGG enrichment analysis were established by to obtain the targets of ZGY against ischemic stroke.2.Bilateral carotid artery ligation was used to establish cerebral ischemia reperfusion model,which was divided into sham,model,EGb,ZGY high,middle and low dose group.Behavioural tests such as open field experiment and rotating rod experiment were carried out before and after administration.Serum biochemical indexes were detected after samples were taken.Brain tissue sections were stained with HE and Nissl,and pathological changes in cerebral cortex and hippocampus were observed.Histochemical preliminary study on the expression of p-NF-?B p65 protein in the brain tissue of mice in each group.3.Cell injury model was established by oxygen glucose deprivation/reperfusion.The total RNA and protein were extracted to determine the expression of p-NF-?B p65 protein,and the expression of key genes NF-?B and IKK? in the NF-?B signaling pathway,and the expression of inflammatory factors TNF-? and iNOS.Results:1.There were 10 common target genes of ZGY related ischemic stroke,and the main chemical components are ellagic acid and apigenin.The core protein network include JUN,HIF1A,RELA,NFKB1A,CDKN1 A,FOS.GO and KEGG enrichment analysis showed that ZGY can treat ischemic stroke through oxidative stress,immune response,and inflammation.2.ZGY can significantly improve cerebral ischemia reperfusion injury.First,improving the exercise ability and autonomous exploration ability of mice,reduce the stress response in the new environment.Secondly,it can regulate the activities of LDH,MDA and SOD to protect cells from free radical attack and reduce the oxidative stress.Finally,it improved the morphology,number and arrangement of neurons in the cortex and hippocampus of the brain.At the same time,ZGY can significantly down-regulate the expression of p-NF-?B p65.3.ZGY inhibited the expression of p-NF-?B p65 protein in OGD/R cells,improved cell survival.The results of RT-PCR showed that ZGY down-regulation the expression of NF-?B and IKK? genes and prevented the activation of NF-?B signaling pathway,at the same time,it can reduce the expression of inflammatory factors iNOS and TNF-? to reduce neuroinflammation.Conclusion:In this paper,network pharmacology technology predicted the targets of ZGY against ischemic stroke,which was supported by animal and cell experiments.On the one hand,ZGY can reduced ischemia damage,on the other hand,it proved that ZGY can inhibit the activation of NF-?B signaling pathway and reduce neuroinflammation,which is of great significance for the follow-up development of ZGY.