Suppression of LPS-induced NF-κB activity in macrophages by the synthetic aurone, (Z)-2-((5-(hydroxymethyl) furan-2-yl) methylene) benzofuran-3(2H)-one

Hyo S Park; David E Nelson; Zachary E Taylor; James B Hayes; Kirsten D Cunningham; Brock A Arivett; Rajarshi Ghosh; Larissa C Wolf; Kimberley M Taylor; Mary B Farone; Scott T Handy; Anthony L Farone

doi:10.1016/j.intimp.2016.12.004

Suppression of LPS-induced NF-κB activity in macrophages by the synthetic aurone, (Z)-2-((5-(hydroxymethyl) furan-2-yl) methylene) benzofuran-3(2H)-one

Int Immunopharmacol. 2017 Feb:43:116-128. doi: 10.1016/j.intimp.2016.12.004. Epub 2016 Dec 16.

Authors

Affiliations

¹ Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: hsp2g@mtmail.mtsu.edu.
² Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: david.e.nelson@mtsu.edu.
³ Middle Tennessee State University, Department of Chemistry, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: zet2d@mtmail.mtsu.edu.
⁴ Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: jhayes2582@gmail.com.
⁵ Middle Tennessee State University, Department of Chemistry, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: kdc7c@mtmail.mtsu.edu.
⁶ Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: baa2j@mtmail.mtsu.edu.
⁷ Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: rg3j@mtmail.mtsu.edu.
⁸ Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: lwolf4@uthsc.edu.
⁹ Middle Tennessee State University, Department of Chemistry, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: kmt4i@mtmail.mtsu.edu.
¹⁰ Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: mary.farone@mtsu.edu.
¹¹ Middle Tennessee State University, Department of Chemistry, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: scott.handy@mtsu.edu.
¹² Middle Tennessee State University, Department of Biology, 1301 East Main Street, Murfreesboro, TN 37132, USA. Electronic address: Anthony.farone@mtsu.edu.

PMID: 27988459
DOI: 10.1016/j.intimp.2016.12.004

Abstract

Suppressing cytokine responses has frequently been shown to have promising therapeutic effects for many chronic inflammatory and autoimmune diseases. However, the severe side effects associated with the long-term use of current treatments, such as allergic reactions and increased risk of stroke, have focused attention towards the targeting of intracellular signaling mechanisms, such as NF-κB, that regulate inflammation. We synthesized a series of non-natural aurone derivatives and investigated their ability to suppress pro-inflammatory signaling in human monocyte (THP-1) and murine macrophage-like (RAW 267.4) cell lines. One of these derivatives, (Z)-2-((5-(hydroxymethyl) furan-2-yl) methylene) benzofuran-3(2H)-one (aurone 1), was found to inhibit LPS-induced secretion of the pro-inflammatory cytokines, tumor-necrosis factor α (TNFα), interleukin 1β (IL-1β), and IL-8 by THP-1 cells. To investigate the mechanism, we probed the effect of aurone 1 on LPS-induced MAPK and NF-κB signaling in both THP-1 and RAW264.7. While aurone 1 pre-treatment had no effect on the phosphorylation of ERK, JNK, or p38 MAPK, it strongly suppressed activation of IKK-β, as indicated by attenuation of Ser176/180 phosphorylation, resulting in decreased phosphorylation of p65 (ser536) as well as phosphorylation (ser32) and degradation of IκBα. Consistent with this, aurone 1 significantly reduced LPS-stimulated nuclear translocation of p65-containing NF-κB transcription factors and expression of an mCherry reporter of TNFα gene transactivation in RAW264.7 cells. Inhibition of TNFα expression at the transcription level was also demonstrated in THP-1 by qRT-PCR. In addition to its effects on cytokine expression, aurone 1 pre-treatment decreased expression of iNOS, a bona fide NF-κB target gene and marker of macrophage M1 polarization, resulting in decreased NO production in RAW264.7 cells. Together, these data indicate that aurone 1 may have the potential to function as a pharmacological agent for the treatment of chronic inflammation disorders.

Keywords: Anti-inflammatory; Aurone; IKK-β; Macrophage; NF-κB; iNOS.

MeSH terms

Animals
Anti-Inflammatory Agents / therapeutic use*
Benzofurans / chemical synthesis
Benzofurans / pharmacology*
Cytokines / metabolism
Humans
Inflammation / drug therapy*
Inflammation Mediators / metabolism
Lipopolysaccharides / immunology
Macrophages / drug effects*
Macrophages / immunology
Mice
Monocytes / drug effects*
Monocytes / immunology
NF-kappa B / metabolism*
Nitric Oxide Synthase Type II / genetics
Nitric Oxide Synthase Type II / metabolism
RAW 264.7 Cells
Signal Transduction / drug effects
Tumor Necrosis Factor-alpha / genetics
Tumor Necrosis Factor-alpha / metabolism*

Substances

Anti-Inflammatory Agents
Benzofurans
Cytokines
Inflammation Mediators
Lipopolysaccharides
NF-kappa B
Tumor Necrosis Factor-alpha
aurone
Nitric Oxide Synthase Type II
Nos2 protein, mouse