Molecular and cellular pharmacologyMethyl (E)-(3-(3,4-dihydroxyphenyl)acryloyl)tryptophanate can suppress MCP-1 expression by inhibiting p38 MAP kinase and NF-κB in LPS-stimulated differentiated THP-1 cells
Graphical abstract
Introduction
Monocyte chemotactic factor-1 (MCP-1) is a small but potent chemotactic factor belonging to the subfamily of CC chemokines (Deshmane et al., 2009, Leonard and Yoshimura, 1990). Its expression is reported to be ubiquitous in many different cell types and is often up-regulated by various stimuli including lipopolysaccharide (LPS), IL-1, TNF-alpha, and 2-O-tetradecanoylphorbol 13-acetate (TPA) (Kunkel et al., 1991, Martin et al., 1997). Because MCP-1 has a strong chemotactic activity on inflammatory cells, it can recruit blood monocytes into the vessel wall, thereby leading to systemic vascular inflammation commonly observed in cardiovascular disease (CVD) and other diseases (Fox and Kahn, 2005, Christodoulidis et al., 2014, Lin et al., 2014, Ruster and Wolf, 2008). Although the regulation of MCP-1 expression is considered complex and cell-specific, p38 MAP kinase and NF-κB are considered major regulatory molecules in the production of MCP-1 in cells (Sutcliffe et al., 2009, Yang et al., 2014, Saklatvala, 2004, Lee et al., 1999). Therefore, the inhibition of these molecules has been suggested as an effective means for suppressing MCP-1 production (Sheryanna et al., 2007, Wong et al., 2005).
Methyl (E)-(3-(3,4-dihydroxyphenyl)acryloyl)tryptophanate (MHAT) is an O-methyl ester of javamide-II which is a phenolic amide found in coffee (Park, 2016). During the course of our study, MHAT was found to have stronger anti-inflammatory activity than the parent chemical javamide-II, speculating its potential to inhibit the expression of inflammation-related molecules including MCP-1 in cells. However, there is currently little information about the capability of MHAT to inhibit p38 MAP kinase, NF-κB and MCP-1 expression in monocytic/macrophage-like cells. Therefore, in this study, MHAT was synthesized, and its potential effect on the expression of MCP-1 was investigated in LPS-stimulated differentiated THP-1 cells by examining its effects on p38 MAP kinase/ATF-2 phosphorylation and NF-κB phosphorylation/transcriptional activity. Additionally, the potential effect of MHAT on RANTES was investigated in the same cells because p38 MAP kinase and NF-κB are also significantly involved in the production of RANTES in the cells (Wong et al., 2005).
Section snippets
Materials
Tryptophan, cinnamic acid, dichloromethane, N,N-dimethylformamide, SB203580, Phorbol 12-myristate 13-acetate (PMA) and other chemicals were purchased from Sigma Chemical Co. (St. Louis, MO). THP-1 cells were purchased from ATCC (Manassas, VA). Alpha-tubulin (Catalog no. 2144), ATF-2 (Catalog no. 9226) and phospho-ATF-2 (Catalog no. 5112) antibodies were purchased from Cell Signaling (Danvers, MA, USA)
Chemical synthesis
The synthesis of MHAT was performed using a method described previously (Park, 2012). Briefly,
Results
MHAT (methyl (E)-(3-(3,4-dihydroxyphenyl)acryloyl)tryptophanate) (Fig. 1) was synthesized using the method described in Section 2,and the product was purified by HPLC (Waters, Milford, MA) as described previously (Park, 2012). To confirm the identities of the synthesized products, the sample was analyzed using NMR spectroscopic methods as described in Section 2, (see Supplementary data for NMR data). Based on the NMR data, the structure of the product was determined as that of methyl
Discussion
In this study, MHAT was found to inhibit MCP-1 production in LPS-stimulated differentiated THP cells by inhibiting p38 kinase/ATF-2 phosphorylation as well as NF-κB. MCP-1 is a significant inflammatory chemokine involved in progressive chronic vascular/interstitial inflammation in CVD, chronic kidney disease and other diseases (Christodoulidis et al., 2014, Lin et al., 2014). There are several isoforms (p38α, β, γ and δ) of p38 MAP kinase that are activated by a variety of cellular stresses
Conflict of interest statement
There are no disclosures to make about financial, consulting, and personal relationships linked to this paper.
Acknowledgements
This study was funded by the USDA (Project no. 8040-51000-057-00D).
References (35)
- et al.
Effect of the p38 MAPK inhibitor SB-239063 on lipopolysaccharide-induced psychomotor retardation and peripheral biomarker alterations in rats
Eur. J. Pharmacol.
(2011) - et al.
Effects of p38 mitogen-activated protein kinase inhibition on vascular and systemic inflammation in patients with atherosclerosis
JACC Cardiovasc. Imaging
(2012) - et al.
Tumor necrosis factor alpha (TNF-alpha)-induced RANTES chemokine expression via activation of NF-kappaB and p38 MAP kinase: roles of TNF-alpha in alcoholic liver diseases
J. Hepatol.
(2003) - et al.
p38 mitogen-activated protein kinase inhibitors--mechanisms and therapeutic potentials
Pharmacol. Ther.
(1999) - et al.
Human monocyte chemoattractant protein-1 (MCP-1)
Immunol. Today
(1990) The p38 MAP kinase pathway as a therapeutic target in inflammatory disease
Curr. Opin. Pharmacol.
(2004)- et al.
Molecular mechanisms underlying chemopreventive activities of anti-inflammatory phytochemicals: down-regulation of COX-2 and iNOS through suppression of NF-kappa B activation
Mutat. Res.
(2001) - et al.
Minocycline modulates cytokine and chemokine production in lipopolysaccharide-stimulated THP-1 monocytic cells by inhibiting IκB kinase α/β phosphorylation
Transl. Res.
(2013) - et al.
High glucose accelerates MCP-1 production via p38 MAPK in vascular endothelial cells
Biochem. Biophys. Res. Commun.
(2003) - et al.
The kinetic mechanism of the dual phosphorylation of the ATF2 transcription factor by p38 mitogen-activated protein (MAP) kinase alpha. Implications for signal/response profiles of MAP kinase pathways
J. Biol. Chem.
(2001)