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The p38 MAPK inhibitor SB203580 differentially modulates LPS-induced interleukin 6 expression in macrophages
Qinghai Shi1, Liping Cheng1, Zhengxiang Liu1
1Clinical Laboratory Diagnostic Center, Urumqi General Hospital of PLA, China.
Abstract:
The p38 mitogen-activated protein kinase (MAPK) plays a key role in lipopolysaccharide (LPS)-induced signal transduction pathways that lead to inflammatory cytokine synthesis in macrophages; however, whether the inhibition of p38 MAPK regulates LPS-induced inflammatory cytokine expression in different types of macrophages remains the subject of debate. Herein, we assessed whether the inhibition of p38 MAPK by SB203580 regulates LPS-induced expression of the inflammatory cytokines tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) in RAW264.7 and resident peritoneal macrophages. Lipopolysaccharide stimulation of RAW264.7 macrophages or mouse resident peritoneal macrophages significantly increased TNF-α and IL-6 production. The addition of SB203580 to cultures dramatically blocked LPS-induced TNF-α production in RAW264.7 and mouse resident peritoneal macrophages, and dramatically blocked LPS-induced IL-6 production in RAW264.7 macrophages, but not in mouse resident peritoneal macrophages. Additionally, high concentrations of SB203580 resulted in increased IL-6 production. However, LPS-stimulation significantly up-regulated the mRNA transcript levels of TNF-α and IL-6 in RAW264.7 and mouse resident peritoneal macrophages, whereas pretreatment with SB203580 dramatically down-regulated LPS-induced mRNA transcript levels of TNF-α and IL-6 in these cells. Our data show that SB203580 differentially modulates LPS-induced production of the inflammatory cytokine IL-6 in two different sources of macrophages, and that this course of regulation occurs at the IL-6 mRNA post-transcriptional stage.
Insights
p38 MAPK inhibition by SB203580 blocks inflammatory cytokine TNF-α production in macrophages. However, IL-6 regulation differs between macrophage types, occurring post-transcriptionally.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- p38 mitogen-activated protein kinase (MAPK) is crucial for inflammatory responses.
- Lipopolysaccharide (LPS) triggers inflammatory cytokine synthesis in macrophages via p38 MAPK signaling.
- The precise role of p38 MAPK inhibition in regulating LPS-induced cytokines across different macrophage types is debated.
Purpose of the Study:
- To investigate how p38 MAPK inhibition affects LPS-induced tumor necrosis factor α (TNF-α) and interleukin 6 (IL-6) expression.
- To compare the effects of SB203580, a p38 MAPK inhibitor, on RAW264.7 and resident peritoneal macrophages.
- To elucidate the regulatory stage of IL-6 expression modulated by p38 MAPK inhibition.
Main Methods:
- Stimulation of RAW264.7 and mouse resident peritoneal macrophages with LPS.
- Treatment with SB203580, a p38 MAPK inhibitor, at various concentrations.
- Quantification of TNF-α and IL-6 protein and mRNA levels using established assays.
- Analysis of post-transcriptional regulation mechanisms.
Main Results:
- LPS significantly increased TNF-α and IL-6 production and mRNA levels in both macrophage types.
- SB203580 effectively blocked LPS-induced TNF-α production and mRNA levels in both macrophage types.
- SB203580 blocked LPS-induced IL-6 production in RAW264.7 cells but not resident peritoneal macrophages.
- High SB203580 concentrations increased IL-6 production, and regulation occurred at the post-transcriptional level for IL-6.
Conclusions:
- SB203580 differentially regulates LPS-induced IL-6 production in RAW264.7 and resident peritoneal macrophages.
- p38 MAPK inhibition impacts inflammatory cytokine expression differently depending on macrophage source.
- Regulation of IL-6 by SB203580 occurs at the post-transcriptional level, highlighting complex signaling pathways.
