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Published on: May 31, 2018
Vasopressin inhibits mitogen-activated protein kinases and activated protein-1 in macrophages
Yu-Long Chen1, Ya-Ying Chang2, Ming-Chang Kao2
1Department of Emergency Medicine, Taipei Tzu Chi Hospital, Taipei, Taiwan.
Objectives:
We have previously shown that vasopressin could inhibit the upregulation of inflammatory mediators. Expression of inflammatory mediators is tightly regulated by the upstream transcriptional pathway mitogen-activated protein kinases (MAPKs) and activated protein-1 (AP-1). In this study, we elucidated whether vasopressin could inhibit the upregulation of MAPKs/AP-1.
Methods:
Murine macrophages (RAW264.7 cells) randomly received lipopolysaccharide (LPS; 100 ng/mL) or LPS plus vasopressin (1000 pg/mL) (designated as the LPS and the LPS+V groups, respectively). Control groups were run simultaneously. For MAPKs, cells were harvested at 0 minutes, 15 minutes, 30 minutes, 45 minutes, and 60 minutes after reaction. For AP-1, cells were harvested at 60 minutes after reaction. Between-group differences in MAPKs (i.e., extracellular regulated kinase, c-Jun N-terminal kinase, and p38 MAPK) and AP-1 expressions were compared.
Results:
Immunoblotting assay data revealed that extracellular regulated kinase concentrations of the LPS+V group that harvested at 45 minutes and 60 minutes, but not at 15 minutes and 30 minutes, were significantly lower than those of the LPS group (p=0.005 and p=0.013). C-Jun N-terminal kinase concentrations of the LPS+V group that harvested at 15 minutes, 30 minutes, 45 minutes, and 60 minutes were also significantly lower than those of the LPS group (all p<0.001). Concentrations of p38 MAPK of the LPS+V group that harvested at 15 minutes, 30 minutes, and 45 minutes, but not at 60 minutes, were also significantly lower than those of the LPS group (all p<0.001). In addition, immunohistochemistry assay revealed that the AP-1 fluorescence signals of the LPS+V group were weaker than those of the LPS group.
Conclusion:
Vasopressin inhibits MAPKs and AP-1 in endotoxin-activated macrophages.
Insights
Vasopressin effectively inhibits the upregulation of mitogen-activated protein kinases (MAPKs) and activated protein-1 (AP-1) in macrophages stimulated with lipopolysaccharide (LPS). This finding suggests vasopressin
Area of Science:
- Immunology
- Cell Biology
- Pharmacology
Background:
- Inflammatory mediator expression is regulated by mitogen-activated protein kinases (MAPKs) and activated protein-1 (AP-1).
- Vasopressin has demonstrated potential in inhibiting inflammatory mediator upregulation previously.
Purpose of the Study:
- To investigate whether vasopressin can inhibit the upregulation of MAPKs and AP-1 in activated macrophages.
- To elucidate the molecular mechanisms underlying vasopressin's anti-inflammatory effects.
Main Methods:
- Murine macrophages (RAW264.7 cells) were stimulated with lipopolysaccharide (LPS) alone or with LPS plus vasopressin.
- Cells were harvested at various time points (0-60 minutes) for analysis of MAPKs (ERK, JNK, p38) and AP-1 expression.
- Immunoblotting and immunohistochemistry assays were employed to quantify protein and signal expressions.
Main Results:
- Vasopressin significantly reduced extracellular regulated kinase (ERK) levels at 45 and 60 minutes post-LPS stimulation.
- C-Jun N-terminal kinase (JNK) concentrations were significantly decreased by vasopressin at all measured time points (15-60 minutes).
- p38 MAPK levels were significantly lower in the vasopressin-treated group at 15, 30, and 45 minutes, and AP-1 signals were weaker.
Conclusions:
- Vasopressin demonstrates an inhibitory effect on MAPKs and AP-1 pathways in macrophages activated by endotoxin.
- These findings highlight vasopressin as a potential therapeutic agent for inflammatory conditions mediated by these pathways.
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