Distinct patterns of intracerebral hemorrhage‐induced alterations in NF‐κB subunit, iNOS, and COX‐2 expression

Transcription factor nuclear factor‐κB (NF‐κB), plays a key role in regulating inflammation in brain pathologies. The goal of this study was to characterize temporal changes in NF‐κB activation, NF‐κB subunit expression, and expression of selected NF‐κB‐regulated gene products [inducible form of nitric oxide synthase (iNOS) and cyclooxygenase‐2], at the transcriptional and translational level in the brain after intracerebral hemorrhage (ICH). Employing the intrastriatal injection of autologous blood in rats to model ICH, we demonstrated, using NF‐κB–DNA binding assay, a robust and prolonged NF‐κB activation, starting as early as 15 min after the onset of ICH. Consequently, we demonstrated that the mRNA and protein for p50, p52, p65, c‐Rel, and RelB NF‐κB subunits, as well as IκBα were all up‐regulated, with a time course ranging from minutes to days following ICH, depending on the subunit. Using reverse transcription‐polymerase chain reaction to analyze mRNA and immunoblotting to analyze protein in ICH‐affected tissue, we found robust induction of iNOS at both mRNA and protein levels that followed a time‐course similar to changes in p65, p52, and RelB mRNA. Oddly, in contrast to iNOS, cyclooxygenase‐2 mRNA and protein following an early transient increase demonstrated significant reduction in response to ICH. In summary, NF‐κB activation occurs within minutes and persists for at least a week in response to ICH. This reaction utilizes different NF‐κB regulatory subunits and is associated with the expression of selected target genes.

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