Bioinformatics Analysis Of The Sensitization Mechanisms Andmolecular Targets Of Echinococcus Granulosus

Objective: To analyze the sensitization mechanism and molecular target of Echinococcus granulosus by bioinformatics.Methods: Nine female BALB/c mice were randomly divided into three groups,each containing three mice.The first group was the healthy control group.At the beginning of the experiment,normal saline(0.2ml/animal)was injected intraperitoneally.After 3 months of feeding,the normal saline for injection was continued to be injected intraperitoneally(0.2ml/animal);The second group was the non-sensitized group with hydatid infection.At the beginning of the experiment,the original scolex suspension was injected intraperitoneally(0.2ml/animal),and the normal saline was injected intraperitoneally(0.2ml/animal)three months later;The third group is the hydatid infection sensitization group.At the beginning of the experiment,the same as the non-sensitized group,the original scolex suspension was used for intraperitoneal injection(0.2ml/animal).After 3 months,the crude capsule solution was extracted by intraperitoneal injection for sensitization(0.2ml/animal).Observe each group for 1 hour after the injection,and then take out the spleen tissue of each experimental group for RNA sequence analysis,find the differentially expressed genes through bioinformatics analysis,use edger(quantitative by RNA-Seq by expectation-Maximization RSEM)software for expression difference significance analysis,use String database and Cytoscape software for network interaction analysis of the differentially expressed genes,and set the score to>500;Use cyto Hubba to obtain the degree of nodes,and filter nodes with a degree greater than 3 for display;DAVID online enrichment analysis tool is used for enrichment analysis,and the analysis results are visualized using R language ggplot2.Results: Compared with the healthy control group,there were 743 differentially expressed genes in the infection sensitization group,including 568 up-regulated genes and 175 down-regulated genes,and 458 specific expressed genes in the two groups.Compared with the healthy control group,the infected non-sensitized group had 554 differentially expressed genes,including 393 up-regulated genes and 161 down-regulated genes,and 269 specifically expressed genes.There are 285 expressed genes in the three groups.Compared with the healthy control group,there are 458 unique genes in the three parts of the differential genes: the hydatid infection sensitization group;Compared with the healthy control group,there are 269 unique genes in the non-sensitized group infected with hydatid and 285 genes in the non-sensitized group infected with hydatid and the non-sensitized group infected with hydatid.The PPI network is constructed by using the string database,and the degree of each node is calculated by using cytohubba,and the nodes with a degree greater than 3 are screened.The larger the degree,the redder the color,indicating the more core in the network,as shown in Figure 2.In the PPI analysis results of the specific genes of the hydatid infection sensitization group and the healthy control group,the core positions include IL-10,IL-6,ptgs2,Ccl1,Cxcl1,Cxcl2,Cxcl3,etc;The non-sensitized group of hydatid infection and the healthy control group included Mpo,Elane,Cyp3a25,etc;The analysis results of the common genes of the hydatid infection sensitization group and the hydatid infection non-sensitization group include Mmp9,Timp1,IL-13,Arg1,Hp,Plg,etc.According to GO enrichment,147,10 and 43 genes were expressed in biological process,cell composition,molecular structure and other aspects of the 458 genes specific to the sensitization group and the control group;The co-expressed gene GO enrichment showed that 56,23 and 5 genes were expressed respectively;Compared with the healthy group,there were 51,13 and 18 genes expressed in the sensitized group.Compared with the healthy control group,the functions of the specific expression genes in the infection sensitization group mainly focus on inflammatory response,immune response,IL-1 cell response,etc;The functions of common genes mainly focus on neuropeptide signaling pathway,IL-1 cell response,and cell response to TNF-α And so on.Compared with the healthy control group,the functions of specific genes in the infected non-sensitized group mainly focus on the nucleosome assembly,the positive regulation of extracellular regulated protein kinase-1(ERKI)and ERK2 cascade.The specific genes of the sensitization group are mainly reflected in 32 pathways,including cytokine interaction,signal transduction and transcriptional activation,including 9 pathways including phosphatidylinositol 3-kinase-protein kinase B signal pathway,complement and coagulation cascade.The specific expression of the non-sensitized group is reflected in 12 pathways,including tyrosine metabolism and linoleic acid metabolism.By selecting the number of different gene pathways,the enrichment results of differentially expressed genes in the hydatid infection sensitized group and the infected non-sensitized group were displayed.Conclusion: 1)Inflammatory reaction plays an important role in the sensitization of echinococcus;2)The high expression of IL-6 may promote the occurrence of inflammation,while IL-10,as an anti-inflammatory factor,may antagonize the occurrence of inflammation;3)IL-13 may be the key gene for immune tolerance after hydatid infection;4)ERK signaling pathway may play an immunomodulatory role in the growth of hydatid;Cytokinecytokine receptor interaction pathway,phosphatidylinositol 3 kinase-protein kinase B signal pathway,and Jak-STAT signal pathway were significantly related to the sensitization of hydatid.