Construction Of Mi Rna Regulatory Network In Hippocampal Neuroinflammation Of Niemann-pick Disease Type C1 Mouse

BackgroundNeuroinflammation is one of the important factors that cause the nervous system damage of Nieman-Pick disease type C1(NPC1),and plays an important role in the occurrence and development of NPC1.Activation of microglia and astrocytes was observed in the brains of NPC1 patients and Npc1-/-mice.The activated glial cells released and secreted a large number of inflammatory factors to participate in the development of neuroinflammation,which eventually caused neuron damage.mi RNAs can participate in the control of neuroinflammation by regulating the expression of inflammation-related genes.However,it is unclear whether mi RNA plays a regulatory role in neuroinflammation of NPC1.ObjectiveAnalysis the changes of neuroinflammatory factors and mi RNA expression in the hippocampus of Npc1-/-mice,constructed a regulatory network of mi RNA-m RNA,to explore the regulatory effect of mi RNA on neuroinflammation,provides a theoretical basis for further elucidating the pathogenesis and immune mechanism of NPC1 and finding the target of drug treatment.MethodsUsing protein chip(RayBiotech,QM-INF-1-1)and Small RNA sequencing to analyze hippocampal tissues of P40 Npc1-/-mice(n=3)and Npc1+/+ mice(n=3),ascreening for differentially expressed inflammatory factors in Npc1-/-mice.Two-way prediction and negative correlation analysis of differentially expressed inflammatory factors and differential mi RNAs were performed by mi RWalk.Finally,a network of mi RNAs to regulate neuroinflammation was constructed through Cytoscape.Results1.The protein chip analysis screened out 12 differentially expressed inflammatory factors(|log2(fold change)|?1,P<0.05),of which 5 were up-regulated and 7 were down-regulated.Protein interaction diagrams showed that there exist interactions among inflammatory factors.A total of 1610 target mi RNAs with 12 differentially expressed inflammatory factors were predicted by mir Walk,among which 5 inflammatory factors were up-regulated.In this study,5 differentially expressed inflammatory factors with up-regulated expression were predicted by mi RWalk to obtain target 932 potential mi RNAs,and 7 down-regulated differential inflammatory factors to target 678 potential mi RNAs.2.Small RNA sequencing identified 83 differentially expressed mi RNAs(|log2(fold change)|?1,P <0.05),of which 42 were up-regulated and 41 were down-regulated.mi RWalk reversely predicts the target m RNA corresponding to 83 mi RNAs differentially expressed in Small RNA results,a total of 26932 target m RNAs,of which 42 differentially up-regulated mi RNAs regulate 15561 target genes,41 differentially down-regulated differential mi RNAs regulate 11374 target genes.3.Draw Venn Diagram database was used to analyze the predicted target mi RNA and Small RNA,target m RNA and protein chip results respectively.As a result,4inflammatory factors up-regulated,4 inflammatory factors down-regulated,and 17up-regulated mi RNA,12 down-regulated mi RNAs.4.Cytoscape constructed 17 regulatory networks that up-regulated mi RNAs regulate4 down-regulated inflammatory factors and 12 down-regulated mi RNAs regulate 4up-regulated inflammatory factors.Important target genes regulated by them include Eotaxin-2,GM-CSF,IFN-gamma,IL-1 beta,ICAM-1,MCP-5,MIP-1 gamma,and TNF RII.ConclusionThrough the combined analysis of protein chip and Small RNA sequencing,a mi RNA network regulating neuroinflammation was constructed,and it was found that mi R-34 a regulates ICAM-1,mi R-219a-2-3p,and mi R-532-3p regulate MCP-5 as regulation The key node of the network,this will provide new ideas and drug treatment targets for NPC1 treatment.