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Objective:To screen differentially expressed proteins(DEPs)in the hippocampus of epileptic rats,pro-viding ideas for further exploring the pathogenesis and drug therapeutic targets of epilepsy.Methods:The epileptic rat model(PTZ group)induced by pentylenetetrazol(PTZ).The protein profiles of hippocampal tissues of the PTZ group and control groups were detected by isobaric tag for relative and absolute quantitation(iTRAQ)combined with LC-MS/MS technology,and the fold changes in the protein expression of the PTZ group versus the control group was considered to be more than 1.5 or less than 0.67,and P＜0.05 was taken as the criteria for screening DEPs.Then DEPs were subjected to bioinformatics analyses such as Gene Ontology(GO)functional annotation and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis.Results:A total of 80 proteins showed significantly different expression,which included 39 up-regulated and 41 down-regulated.GO analysis showed that the up-regulated DEPs were mainly involved in biological processes such as cellular response to nerve growth factor stimulus,axon develop-ment,cell surface receptor signaling pathway,neuron migration,actin filament depolymerization,and signal transduc-tion,while down-regulated DEPs were mainly involved in biological processes such as tricarboxylic acid cycle,citrate metabolism process,acetyl-CoA biosynthetic process from pyruvate,oxaloacetate metabolism process,and regulation of focal adhesion assembly.KEGG pathway enrichment analysis showed that up-regulated DEPs were mainly involved in five signaling pathways,including AMPK signaling pathway,regulation of actin cytoskeleton,sphingolipid signaling pathway,phenylalanine metabolism,and insulin signaling pathway;The down-regulated DEPs were mainly involved in six signaling pathways,including the citric acid cycle,carbon metabolism,acetate and dicarboxylate metabolism,meta-bolic pathways,amino acid biosynthesis and amyotrophic lateral sclerosis signaling pathways.Conclusion:DEPs from epileptic hippocampal tissues were screened by iTRAQ proteomics,and the metabolic pathways enriched by bioinformat-ics methods for DEPs may be closely related to the pathogenesis of epilepsy.
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Objective To investigate the expression of neuron-specific enolase(NSE) and bone morphogenic protein 4(BMP4) in different hippocampal areas of pentylenetetrazol(PTZ) kindled epilepsy rats and explore their relationship with the pathogenesis of epilepsy and brain injury.Methods Fifty male SD rats were divided into experimental group(n=40) and control group(n=10).The rats in experimental group were kindled into epilepsy by chemical method,and according to the kindling process,subdivided into four groups(grade Ⅰ,Ⅲ,Ⅳ,Ⅴ).Immunohistochemistry,in situ hybridization labeled with Dig-oligonucleotide probe and the image analyzing system were used to observe the expressions of NSE and BMP4 in rat hippocampus.Results In PTZ kindled epilepsy rats,the number of cells positive for NSE and BMP4 was increased in many regions of hippocampal formation.Compared with control group,the expressions of NSE and BMP4 in CA3 and DG was elevated obviously in the grade Ⅲ group and grade Ⅳ group(P
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Objective To study the role of GABA (? aminobutyric acid) transporters in kindled.Methods The rat's GABA model following PTZ kindled was made.The expression of GABA transporter subtype in the hippocampus(GAT 1 mRNA) was measured after PTZ (pentylenetetrazol) kindled seizures by using semiquantitative technique,reverse transcription polymerase chain reaction (RT PCR).Results GAT 1 mRNA was significantly increased in the hippocampus at 0 hour,48 hours,and 1 week after PTZ kindled seizures. Up regulation of expression of GAT 1 mRNA returned to the control levels after 1 month. No changes in the expression of GAT 1 mRNA were observed after 60 days of PTZ kindled seizures.Conclusion The expression of GAT 1 mRNA with up regulation may be associated with the epilepsy susceptibility.