Downregulation of the NHE3-Binding PDZ-Adaptor Protein PDZK1 Expression during Cytokine-Induced Inflammation in Interleukin-10–Deficient Mice 英文参考文献.docVIP

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Downregulation of the NHE3-Binding PDZ-Adaptor Protein PDZK1 Expression during Cytokine-Induced Inflammation in Interleukin-10–Deficient Mice 英文参考文献.doc

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Downregulation of the NHE3-Binding PDZ-Adaptor Protein PDZK1 Expression during Cytokine-Induced Inflammation in Interleukin-10–Deficient Mice 英文参考文献

DownregulationoftheNHE3-BindingPDZ-Adaptor ProteinPDZK1ExpressionduringCytokine-Induced InflammationinInterleukin-10–DeficientMice HenrikeLenzen1*,MariaLu¨nnemann1,AndreBleich2,MichaelPManns1,UrsulaSeidler1,AnneJo¨rns3,4 1DepartmentofGastroenterology,HepatologyandEndocrinology,HannoverMedicalSchool,Hannover,Germany,2InstituteofLaboratoryAnimalScience,Hannover MedicalSchool,Hannover,Germany,3InstituteofClinicalBiochemistry,HannoverMedicalSchool,Hannover,Germany,4CenterofAnatomy,HannoverMedicalSchool, Hannover,Germany Abstract Background:Impairedsaltandwaterabsorptionisanimportantfeatureinthepathogenesisofdiarrheaininflammatory bowel disease (IBD). We analyzed the expression of proinflammatory cytokines in the infiltrating immune cells and the + + function and expression of the Na /H exchanger isoform 3 (NHE3) and its regulatory PDZ-adaptor proteins NHERF1, NHERF2,andPDZK1inthecolonofinterleukin-10–deficient(IL-102/2 )mice. Methodology/Principal Findings: Gene and protein expression were analyzed by real-time reverse transcription polymerase chain reaction (qRT-PCR), in situ RT-PCR, and immunohistochemistry. NHE3 activity was measured fluorometrically in apical enterocytes within isolated colonic crypts. Mice developed chronic colitis characterized by a typical immune cell infiltration composed of T-lymphocytes and macrophages, with high levels of gene and protein expressionoftheproinflammatorycytokinesinterleukin-1bandtumornecrosisfactor-a.Inparallel,induciblenitricoxide synthaseexpressionwasincreasedwhileprocaspase3expressionwasunaffected. Interferon-cexpressionremainedlow. Although acid-activated NHE3 activity was significantly decreased, the inflammatory process did not affect its gene and protein expression or its abundance and localization in the apical membrane. However, expression of the PDZ-adaptor proteins NHERF2 and PDZK1 was downregulated. NHERF1 expression was unchanged. In a comparative analysis we observedthePDZK1downregulationalsointheDSS(de