The Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 regulate the levels of intracellular chloride in hippocampal cells. Impaired chloride transport by these proteins is thought to be involved in the pathophys...The Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 regulate the levels of intracellular chloride in hippocampal cells. Impaired chloride transport by these proteins is thought to be involved in the pathophysiological mechanisms of mesial temporal lobe epilepsy. Imbalance in the relative expression of these two proteins can lead to a collapse of CI- homeostasis, resulting in a loss of gamma-aminobutyric acid-ergic inhibition and even epileptiform discharges. In this study, we investigated the expression of Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 in the sclerosed hippocampus of patients with mesial temporal lobe epilepsy, using western blot analysis and immunohistochemistry. Compared with the histologically normal hippocampus, the sclerosed hippocampus showed increased Na＋-K＋-Cl- cotransporter 1 expression and decreased K＋-CI- cotransporter 2 expression, especially in CA2 and the dentate gyrus. The change was more prominent for the Na＋-K＋-CI- cotransporter 1 than for the K＋-CI- cotransporter 2. These experimental findings indicate that the balance between intracellular and extracellular chloride may be disturbed in hippocampal sclerosis, contributing to the hyperexcitability underlying epileptic seizures. Changes in Na＋-K＋-CI-cotransporter 1 expression seems to be the main contributor. Our study may shed new light on possible therapies for patients with mesial temporal lobe epilepsy with hippocampal sclerosis.展开更多
Background Autophagy plays essential roles in the development and pathogenesis of mesial temporal lobe epilepsy(mTLE).In this research,we aim to identify and validate the autophagy-related genes associated with mTLE t...Background Autophagy plays essential roles in the development and pathogenesis of mesial temporal lobe epilepsy(mTLE).In this research,we aim to identify and validate the autophagy-related genes associated with mTLE through bioinformatics analysis and experimental validations.Methods We obtained the dataset GSE143272 and high-throughput sequencing results of mTLE from public data-bases.Potential differentially expressed autophagy-related genes related to mTLE were identified using R software.Subsequently,genomes pathway enrichment analysis,protein-protein interactions(PPIs),and the gene ontology(GO)enrichment were performed for the selected autophagy-related genes.The mRNA expression profiles of hub genes were then used to establish a least absolute shrinkage and selection operator(LASSO)model.Finally,seven hub candidate autophagy-related genes were confirmed in hippocampus using the lithium-pilocarpine chronic epilepsy model.Results A total of 40 differential expression genes(DEGs)among the core autophagy-related genes were identified.The analysis results of PPI revealed that interactions among these DEGs.KEGG pathway and GO analysis of selected candidate autophagy-related genes indicated that those enriched terms mainly focused on macroautophagy,regula-tion of autophagy,cellular response to extracellular stimulus and mitochondrion disassembly.The results suggested that SQSTM1,VEGFA,BNIP and WIPI2 were consistent with the bioinformatics analysis.The expression levels of SQSTM1 and VEGFA in epilepsy model samples were significantly higher than those in normal control,while BNIP and WIPI2 expression levels were notably decreased.The final hub gene-based LASSO regression model accurately predicted the occurrence of epilepsy(AUC=0.88).Conclusions Through bioinformatics analysis of public data,we identified 40 candidate autophagy-related genes associated with mTLE.SQSTM1,VEGFA,BNIP and WIPI2 may play significant roles in autophagy,influencing the onset and development of mTLE by regulating autophagy pathway.These findings deepen our understanding of mTLE,and may serve as sensitive and valuable indicators for the prognosis and diagnosis of this condition.展开更多
Background:This study was designed to characterize human PRRT2 gene and protein,in order to provide theoretical reference for research on regulation of PRRT2 expression and its involvement in the pathogenesis of parox...Background:This study was designed to characterize human PRRT2 gene and protein,in order to provide theoretical reference for research on regulation of PRRT2 expression and its involvement in the pathogenesis of paroxysmal kinesigenic dyskinesia and other related diseases.Method:Biological softwares Protparam,Protscale,MHMM,SignalP 5.0,NetPhos 3.1,Swiss-Model,Promoter 2.0,AliBaba2.1 and EMBOSS were used to analyze the sequence characteristics,transcription factors of human PRRT2 and their binding sites in the promoter region of the gene,as well as the physicochemical properties,signal peptides,hydrophobicity property,transmembrane regions,protein structure,interacting proteins and functions of PRRT2 protein.Results:(1)Evolutionary analysis of PRRT2 protein showed that the human PRRT2 had closest genetic distance from Pongo abelii.(2)The human PRRT2 protein was an unstable hydrophilic protein located on the plasma membrane.(3)The forms of random coil(67.65%)and alpha helix(23.24%)constituted the main secondary structure elements of PRRT2 protein.There were also multiple potential phosphorylation sites in the protein.(4)The results of ontology analysis showed that the cellular component of PRRT2 protein was located in the plasma membrane;the molecular function of PRRT2 included syntaxin-1 binding and SH3 domain binding;the PRRT2 protein is involved in biological processes of negative regulation of soluble NSF attachment protein receptor(SNAR E)complex assembly and calcium-dependent activation of synaptic vesicle fusion.(5)String database analysis revealed 10 proteins with close interactions with the human PRRT2 protein.(6)There were at least two promoter regions in the PRRT2 gene within 2000 bp upstream the 5'flank,a 304-bp CpG island in the promoter region and four GC boxes in the 5'regulatory region of PRRT2 gene and we found 13 transcription factors that could bind the promoter region of the PRRT2 gene.Conclusion:These results provide important information for further studies on the role of PRRT2 gene and identify their functions.展开更多
基金supported by the Science and Technology Foundation of Guangdong Province,No.2008B060600063the National Natural Science Foundation of China,No. 81071050the Natural Science Foundation of Guangdong Province,No. S2011020005483
文摘The Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 regulate the levels of intracellular chloride in hippocampal cells. Impaired chloride transport by these proteins is thought to be involved in the pathophysiological mechanisms of mesial temporal lobe epilepsy. Imbalance in the relative expression of these two proteins can lead to a collapse of CI- homeostasis, resulting in a loss of gamma-aminobutyric acid-ergic inhibition and even epileptiform discharges. In this study, we investigated the expression of Na＋-K＋-CI- cotransporter 1 and K＋-CI- cotransporter 2 in the sclerosed hippocampus of patients with mesial temporal lobe epilepsy, using western blot analysis and immunohistochemistry. Compared with the histologically normal hippocampus, the sclerosed hippocampus showed increased Na＋-K＋-Cl- cotransporter 1 expression and decreased K＋-CI- cotransporter 2 expression, especially in CA2 and the dentate gyrus. The change was more prominent for the Na＋-K＋-CI- cotransporter 1 than for the K＋-CI- cotransporter 2. These experimental findings indicate that the balance between intracellular and extracellular chloride may be disturbed in hippocampal sclerosis, contributing to the hyperexcitability underlying epileptic seizures. Changes in Na＋-K＋-CI-cotransporter 1 expression seems to be the main contributor. Our study may shed new light on possible therapies for patients with mesial temporal lobe epilepsy with hippocampal sclerosis.
基金the National Natural Science Foundation of China(82071447,81571266,81771405)the Sanming Project of Medicine in Shenzhen(No.SZSM201911003).
文摘Background Autophagy plays essential roles in the development and pathogenesis of mesial temporal lobe epilepsy(mTLE).In this research,we aim to identify and validate the autophagy-related genes associated with mTLE through bioinformatics analysis and experimental validations.Methods We obtained the dataset GSE143272 and high-throughput sequencing results of mTLE from public data-bases.Potential differentially expressed autophagy-related genes related to mTLE were identified using R software.Subsequently,genomes pathway enrichment analysis,protein-protein interactions(PPIs),and the gene ontology(GO)enrichment were performed for the selected autophagy-related genes.The mRNA expression profiles of hub genes were then used to establish a least absolute shrinkage and selection operator(LASSO)model.Finally,seven hub candidate autophagy-related genes were confirmed in hippocampus using the lithium-pilocarpine chronic epilepsy model.Results A total of 40 differential expression genes(DEGs)among the core autophagy-related genes were identified.The analysis results of PPI revealed that interactions among these DEGs.KEGG pathway and GO analysis of selected candidate autophagy-related genes indicated that those enriched terms mainly focused on macroautophagy,regula-tion of autophagy,cellular response to extracellular stimulus and mitochondrion disassembly.The results suggested that SQSTM1,VEGFA,BNIP and WIPI2 were consistent with the bioinformatics analysis.The expression levels of SQSTM1 and VEGFA in epilepsy model samples were significantly higher than those in normal control,while BNIP and WIPI2 expression levels were notably decreased.The final hub gene-based LASSO regression model accurately predicted the occurrence of epilepsy(AUC=0.88).Conclusions Through bioinformatics analysis of public data,we identified 40 candidate autophagy-related genes associated with mTLE.SQSTM1,VEGFA,BNIP and WIPI2 may play significant roles in autophagy,influencing the onset and development of mTLE by regulating autophagy pathway.These findings deepen our understanding of mTLE,and may serve as sensitive and valuable indicators for the prognosis and diagnosis of this condition.
基金supported by the Sanming Project of Medicine in Shenzhen(No.SZSM201911003)Guangdong Province Medical Science and Technology Research Found project(A2018320)the National Nature Science Foundation of China(No.81571266,82071447 and 81771405).
文摘Background:This study was designed to characterize human PRRT2 gene and protein,in order to provide theoretical reference for research on regulation of PRRT2 expression and its involvement in the pathogenesis of paroxysmal kinesigenic dyskinesia and other related diseases.Method:Biological softwares Protparam,Protscale,MHMM,SignalP 5.0,NetPhos 3.1,Swiss-Model,Promoter 2.0,AliBaba2.1 and EMBOSS were used to analyze the sequence characteristics,transcription factors of human PRRT2 and their binding sites in the promoter region of the gene,as well as the physicochemical properties,signal peptides,hydrophobicity property,transmembrane regions,protein structure,interacting proteins and functions of PRRT2 protein.Results:(1)Evolutionary analysis of PRRT2 protein showed that the human PRRT2 had closest genetic distance from Pongo abelii.(2)The human PRRT2 protein was an unstable hydrophilic protein located on the plasma membrane.(3)The forms of random coil(67.65%)and alpha helix(23.24%)constituted the main secondary structure elements of PRRT2 protein.There were also multiple potential phosphorylation sites in the protein.(4)The results of ontology analysis showed that the cellular component of PRRT2 protein was located in the plasma membrane;the molecular function of PRRT2 included syntaxin-1 binding and SH3 domain binding;the PRRT2 protein is involved in biological processes of negative regulation of soluble NSF attachment protein receptor(SNAR E)complex assembly and calcium-dependent activation of synaptic vesicle fusion.(5)String database analysis revealed 10 proteins with close interactions with the human PRRT2 protein.(6)There were at least two promoter regions in the PRRT2 gene within 2000 bp upstream the 5'flank,a 304-bp CpG island in the promoter region and four GC boxes in the 5'regulatory region of PRRT2 gene and we found 13 transcription factors that could bind the promoter region of the PRRT2 gene.Conclusion:These results provide important information for further studies on the role of PRRT2 gene and identify their functions.
