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LPS induced alterations in proinflammatory cytokine expression in mouse brain
Author(s) -
Datta Subhash C,
Opp Mark R
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.6.a1393-d
Subject(s) - cytokine , proinflammatory cytokine , lipopolysaccharide , tumor necrosis factor alpha , endocrinology , biology , messenger rna , interleukin 6 , medicine , chemistry , microbiology and biotechnology , inflammation , immunology , biochemistry , gene
Bacterial cell‐wall lipopolysaccharide (LPS) induces peripheral release of interleukin (IL)1β, IL6 and tumor necrosis factor (TNF)α. These cytokines may influence CNS mechanisms regulating sleep by activating neuronal pathways in brain. We previously demonstrated that LPS‐induced fever and increases in sleep are attenuated by inhibition of nuclear translocation of NFkB in mouse brain. To date, there are few studies of the impact of LPS on cytokine expression in discrete mouse brain regions involved in the regulation of sleep. In the present study, we determined cytokine mRNA and protein content in mouse brain after LPS challenge. Adult male mice (~25 g; C57BL/6J) were injected IP at light onset with 10μg LPS (E.coli serotype O111:B4) or vehicle (PFS) and sacrificed 2 or 4 h later. Discrete brain regions were dissected, and tissues frozen and stored at −80°C. RNA was extracted from the 2 h group ( LPS, n=8; PFS, n=8). QPCR (TaqMan; Applied Biosystems) and the comparative threshold cycle (ΔΔCT) method was used to determine cytokine mRNA. LPS increased IL1β, IL6 and TNFα mRNA in hypothalamus, hippocampus and brain stem by 2 to 7 fold. Protein was extracted from the 4 h group for Bio‐Plex assay (BioRad; LPS, n=6, ;PFS, n=6). Cytokine protein in these brain regions increased by 3 to 5 fold. Present results, and our previous data, suggest that LPS‐induced increases in cytokines in brain are involved in alterations in sleep after this challenge. Studies are underway to determine if inhibition of nuclear translocation of NFkB blocks LPS‐induced increases in cytokine protein in mouse brain.