Role of tristetraprolin phosphorylation in paediatric patients with inflammatory bowel disease  

作　　者：Alessia Di Silvestre Marianna Lucafo Letizia Pugnetti Matteo Bramuzzo Gabriele Stocco Egidio Barbi Giuliana Decorti 

机构地区：[1]PhD School in Science of Reproduction and Development,University of Trieste,Trieste 34127,Italy [2]Institute for Maternal and Child Health,IRCCS“Burlo Garofolo”,Trieste 34137,Italy [3]Department of Life Sciences,University of Trieste,Trieste 34127,Italy [4]Department of Medicine,Surgery and Health Sciences,University of Trieste,Trieste 34127,Italy

出　　处：《World Journal of Gastroenterology》2019年第39期5918-5925,共8页世界胃肠病学杂志（英文版）

基　　金：Supported by the Italian Ministry of Health projects Ricerca Corrente1/17 and 21/17(Institute for Maternal and Child Health IRCCS Burlo Garofolo)

摘　　要：BACKGROUND Intestinal inflammation and epithelial injury are the leading actors of inflammatory bowel disease(IBD),causing an excessive pro-inflammatory cytokines expression.Tristetraprolin(TTP),an mRNA binding protein,plays a role in regulating the inflammatory factors,recognizing specific sequences on the 3’untranslated region of cytokine mRNAs.TTP activity depends on its phosphorylation state:the unphosphorylated TTP degrades pro-inflammatory cytokine mRNAs;on the contrary,the phosphorylated TTP fails to destabilize mRNAs furthering their expression.The phospho-TTP forms a complex with the chaperone protein 14-3-3.This binding could be one of the factors that promote intestinal inflammation as a cause of disease progression.AIM To assess if TTP phosphorylation has a role in paediatric IBD.METHODS The study was carried out on a cohort of paediatric IBD patients.For each patient enrolled,a specimen of inflamed and non-inflamed colonic mucosa was collected.Furthermore,the experiments were conducted on macrophages differentiated from blood samples of the same patients.Macrophages from healthy donors’blood were used as controls.Co-immunoprecipitation assay and immunoblotting analyses were performed to observe the formation of the phospho-TTP/14-3-3 complex.In the same samples TNF-αexpression was also evaluated as major factor of the pro-inflammatory activity.RESULTS In this work we studied indirectly the phosphorylation of TTP through the binding with the chaperone protein 14-3-3.In inflamed and non-inflamed colon mucosa of IBD paediatric patients immunoblot assay demonstrated a higher expression of the TTP in inflamed samples respect to the non-inflamed;the coimmunoprecipitated 14-3-3 protein showed the same trend of expression.In the TNF-αgene expression analysis higher levels of the cytokine in inflamed tissues compared to controls were evident.The same experiments were conducted on macrophages from IBD paediatric patients and healthy controls.The immunoblot results demonstrated a high expression of both TBACKGROUND Intestinal inflammation and epithelial injury are the leading actors of inflammatory bowel disease(IBD), causing an excessive pro-inflammatory cytokines expression. Tristetraprolin(TTP), an m RNA binding protein, plays a role in regulating the inflammatory factors, recognizing specific sequences on the 3’ untranslated region of cytokine m RNAs. TTP activity depends on its phosphorylation state: the unphosphorylated TTP degrades pro-inflammatory cytokine m RNAs; on the contrary, the phosphorylated TTP fails to destabilize m RNAs furthering their expression. The phospho-TTP forms a complex with the chaperone protein 14-3-3. This binding could be one of the factors that promote intestinal inflammation as a cause of disease progression.AIM To assess if TTP phosphorylation has a role in paediatric IBD.METHODS The study was carried out on a cohort of paediatric IBD patients. For each patient enrolled, a specimen of inflamed and non-inflamed colonic mucosa was collected.Furthermore, the experiments were conducted on macrophages differentiated from blood samples of the same patients. Macrophages from healthy donors’ blood were used as controls. Co-immunoprecipitation assay and immunoblotting analyses were performed to observe the formation of the phospho-TTP/14-3-3 complex. In the same samples TNF-α expression was also evaluated as major factor of the pro-inflammatory activity.RESULTS In this work we studied indirectly the phosphorylation of TTP through the binding with the chaperone protein 14-3-3. In inflamed and non-inflamed colon mucosa of IBD paediatric patients immunoblot assay demonstrated a higher expression of the TTP in inflamed samples respect to the non-inflamed; the coimmunoprecipitated 14-3-3 protein showed the same trend of expression. In the TNF-α gene expression analysis higher levels of the cytokine in inflamed tissues compared to controls were evident. The same experiments were conducted on macrophages from IBD paediatric patients and healthy controls. The immunoblot results demonstrat

关 键 词：INFLAMMATORY BOWEL disease TRISTETRAPROLIN 14-3-3 protein PHOSPHORYLATION CYTOKINES INFLAMMATION 

分 类 号：R57[医药卫生—消化系统]