Bioinformatics Analysis Of Schizophrenia And Experimental Study Of Infection Model During Pregnancy

Object:Schizophrenia(SZ)is a serious and complex psychiatric syndrome.Several genes that may be involved in SZ have been identified previously,but they are still not completely clear.In this study,we used systematic bioinformatics analysis to integrate two expression datasets,attempting to further identify the related genes and their function along with the key signal pathways,offering references for future research.Methods:1.Two datasets(GSE 17612 and GSE21138)were acquired from the Gene Expression Omnibus(GEO)database.2.Batch normalize was conducted by sva package of R language software and differentially expressed genes(DEGs)were selected by limma package of R language software.3.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were conducted for DEGs.4.Protein-protein interaction(PPI)networks were assembled with Cytoscape software and important moduleswere analyzed by the Molecular Complex Detection(MCODE)algorithm.In addition,the drug-target network of DEGs from important modules was conducted to predict the therapeutic target.Finally,the genes with the most significant differential expression changesscreened out from the important modules were indentified as the hub-genes of schizophrenia.Results:1.259 DEGs were screened out,of which 107 genes were up-regulated(the top five significantly differentially expressed were SST?EGR1?IL-6?CRH and ZNF324)and 152 genes were down-regulated(the top five significantly differentially expressed were FOSB?NPAS4?SCG2?SLN and LBH).2.The DEGs were clustered by analyzing GO and KEGG pathway.In the biological process(BP),the DEGs were mainly enriched in inflammatory response,immune response,neutrophil chemotaxis,leukocyte migration,response to lipopolysaccharide.In cellular component(CC)group,the DEGs were mainly enriched extracellular region,extracellular space,integral component of plasma membrane,secretory granule lumen.In molecular function(MF)group,the DEGs were enriched in transcription factor activity,RNA polymerase ? core promoter proximal region sequence-specific binding,hyaluroniic acid binding,hormone activity,neuropeptide hormone activity.3.A protein-protein interaction(PPI)network of DEGs was constructed which consisted of 155 nodes and 620 edges.IL-6 connected to most node(57 nodes).4.Three most significant modules were selected from the PPI networks.In the three modules,the major biological functionswere involved in inflammatory response,G protein-coupled receptor signaling pathway and response to progesterone stimulation.5.Drug-target network analysis of DEGs showed that TF,IL-6 and S100A8 were the main target genes of drugs labeled in drug-Bank.6.The genes with the most significant differential expression changes were selected from the three important modules,finally,5 hub genes(SST,SCG2,IL-6,CRH,FOSB)were selected,among which IL-6 may be the most pivotal.Conclusion:1.259 DEGs were screened out from 2 microarray expression datasets by integrated bioinformatical analysis.The main biological function of the DEGs was:inflammatory response,immune response,neutrophil chemotaxis,leukocyte migration and response to lipopolysaccharide.2.Five hub genes were screened out,they might be used as the new risk genes of SZ.Especially,IL-6 could be the main potentially target gene for therapeutic drug.Objective:Immune imbalanceplays an important role in the occurrence and development of schizophrenia. A number of studies have confirmed the increased risk of mentalillness in the offspring of bacterial and viralinfections during maternal pregnancy. In the first part of the study,the results suggest that paticnts with immune inflammation and schizophzenia In this part, it is proposed to use the LPSmatemal immune activation model to observe the behavioral change of autonomous activity, learning and memory ability of puberty rats (PND45) and adulthood (PND60) through immune activation of pregnant rats.Thenit is compared with the bchavioral phenotypes of other types of schizophrenia models. In addition, we further observed whether LPS immuneactivation in early pregnancy mice affected the blood-brain barrier integrity of the offspring and the expression of inflammation-related proteins in the nervous system.Method:1.The effect of immune activation of rats in late pregnancy on the behavior of offspeing:First, establish a model of immune activation during pregnancy. SD rats were randomly injected with LPS or saline at 17-19 days of gestation to observe the behaviors of the two groups of rats in adolescence (PND45) and adulchood (PND60)(including open field experiments, new object recognition, Morris water maze).2. Behavioral differences of rats in different schizophrenia models: the social isolation (Isoation, Iso) model,phencyclidine(PCP) model, Mdrl gene knockout model and the previous LPS modelwere used to compare whether the behavioral performance of different model animals varies.3.Detection of blood-brain barrier and inflammatory related proteins in mice with immune activation in early pregnancy C57BL/6 mice were randomly injected intraperitoneally with L.PS or saline on days 7and 8 of pregnancy. The offspring were divided into LPS group and control group (CON) aecoeding to their parental sources. On the postnatal day 7(PND7) and postnatal day 21(PND21), Evans blue staining was used to observe the blood-brain barrier permeability of the two groups; At PND21,western blot analysis was used to analyze the expression of the blood-brain barrier-related proteins Occludin andPDLIM1 in the prefrontal cortex and hippocampus,and the expression of IL-6, CD4and CCL2 in prefrontal cortex was analyzed.Results:1. Behavioral results of each model(1) The total distance of the open field experiment: the total distance of the LPS group during adolescence and adulthood reduced compared with the CON group (P<0.05);the total distance of various types of schizophrenia models (LPS, Iso, Mdrl, PCP) was less than that of CON groups (P<0.05), and there was no significant difference in the total distance of each type of schizophrenia model;(2) Recognition index of new object recognition experiment: in adolescence,the recognition index of LPS group is less than CON group (P=0.016);but in adult,there is no significant difference between these two groups(P=0.3); the recognition index in cach schizophrenia models has no significant difference;(3) Place navigationin water maze: In adolescence, the escape latency of CON and LPS shortened with the extension of days (P<0.05) during the learning stage,but cscape latency of LPS in adultwas not significantly shortened with the extension of days (P> 0.05); There was no significant difference in escape latency between the two groups;and no significant difference in escape latency between each schizophrenia groups [F(4, 30) ? 1.59,P=0.2]. Afterwards, the escape latency in the LPS group was longer than the CON ( P=0.029) by the LSD(least significant difference)pairwise comparison.Spatial probe testin watermaze: the adolescent LPS group stayed in the original quadrant of platform less than the CON group (P=0.013); the adult LPS group residence time in the original quadrant of platform was not statistically different from the CON group (P>0.05); there was no statistical difference in duration time between each model groups.2. Effect of LPS immune activation in carly pregnancy on the expression of BBB and related proteins in the offspring(1) PND7 and PND2I brains in the CON group did not show blue staining; PND7brains in the LPS group had blue staining, while PND21 brains did not show blue staining;(2) At PND21, the expeession of Occludin protein in the prefrontal cortex of the LPS group was higher than that of the CON group (P<0.05), and there was no significant difference in the expression of Occludin in the hippocampus between the two groups; the expression of PDLIM] in the prefrontal cortex and hippocampus between LPS and CON groups showed no significant difference; In the prefrontal cortex, there was no statistically significant difference in the expression of IL-6, CD4, and CCL2 in the LPS group and the CON group (P=0.57, P=0.083,P=0.059), but there is a rising trendencyof CD4 and CCL2 in the LPS group.Conclusion:1. The offspring of LPS immune activation model rats has behavioral abnormalities, especially in adolescence. During adolescence, reduction is observed inthe spontaneous activity, the exploration for the new things and the memory retention ability of the spatial position; During adulthood,it showed that the spoataneous activity and the learning ability decreased; however, there are no changes in BBB permeability of the offspring.2. The LPS immune activation model and other types of schizophrenia models can induce behavioral changes in rats, but the LPS immune activation model rats seems to be more obviously damaged in short-term memory and spatial location cognitive memory;3. The offspring of mice infected with LPS in early pregnancy showed an increase in blood-brain barrier permeability at PND7; no obvious changes in permeability were found at PND21, but the expressicn of Occludin protein in the prefrontal cortex the prefrontal cortex was notstatistically different between the two groups, but CD4and CCL2 tended to increase in the LPS group.