Cuenca Jimena, Goren Nora, Prieto Patricia, Martín-Sanz Paloma, Boscá Lisardo
Instituto de Investigaciones Biomédicas Alberto Sols (Consejo Superior de Investigaciones Cientificas-Universidad Autónoma de Madrid, Spain.
Am J Pathol. 2007 Sep;171(3):820-8. doi: 10.2353/ajpath.2007.061076. Epub 2007 Aug 3.
The ability of neonatal and adult cardiomyocytes to activate the nuclear factor (NF)-kappaB pathway in response to lipopolysaccharide and interleukin-1beta challenge has been investigated and compared with that of peritoneal macrophages. The activation of the IkappaB kinase and the phosphorylation and degradation of IkappaBalpha and IkappaBbeta was much lower in adult cardiomyocytes than in the neonatal counterparts and macrophages. This restricted activation of the NF-kappaB pathway resulted in a significant reduction in the time of nuclear activation of NF-kappaB, as deduced by electrophoretic mobility shift assays and in the transcription of target genes, such as IkappaBalpha, cyclooxygenase-2 (COX-2) and nitric-oxide synthase-2 (NOS-2). Studies on chromatin immunoprecipitation showed binding of NF-kappaB proteins to the regulatory kappaB sites identified in the promoters of the IkappaBalpha, COX-2, and NOS-2 genes in macrophages and, to a lower extent, in neonatal cardiomyocytes. The binding to these kappaB sites in adult cardiomyocytes was observed only in the IkappaBalpha promoter and was minimal or absent in the COX-2 and NOS-2 promoters, respectively, suggesting a restricted activation of NF-kappaB-regulated genes in these cells. These data indicate that the function of the NF-kappaB pathway in adult cardiomyocytes is limited in time, which results in the expression of a reduced number of genes and provides a functional explanation for the absence of NOS-2 inducibility in these cells under proinflammatory conditions.
研究了新生和成年心肌细胞在脂多糖和白细胞介素-1β刺激下激活核因子(NF)-κB通路的能力,并与腹膜巨噬细胞进行了比较。成年心肌细胞中IκB激酶的激活以及IκBα和IκBβ的磷酸化和降解程度远低于新生心肌细胞和巨噬细胞。NF-κB通路的这种受限激活导致通过电泳迁移率变动分析推断的NF-κB核激活时间以及靶基因(如IκBα、环氧化酶-2(COX-2)和一氧化氮合酶-2(NOS-2))转录的显著减少。染色质免疫沉淀研究表明,NF-κB蛋白与巨噬细胞中IκBα、COX-2和NOS-2基因启动子中鉴定的调节性κB位点结合,在新生心肌细胞中的结合程度较低。仅在IκBα启动子中观察到成年心肌细胞与这些κB位点的结合,而在COX-2和NOS-2启动子中分别极少或未观察到结合,这表明这些细胞中NF-κB调节基因的激活受限。这些数据表明,成年心肌细胞中NF-κB通路的功能在时间上受到限制,这导致表达的基因数量减少,并为这些细胞在促炎条件下缺乏NOS-2诱导性提供了功能解释。