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Evidence for inhibition of nitric oxide and inducible nitric oxide synthase in Caco-2 and RAW 264.7 cells by a Maillard reaction product [5-(5,6-dihydro-4H-pyridin-3-ylidenemethyl)furan-2-yl]-methanol

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Abstract

We have recently isolated and characterized the chemical structure of a bioactive Maillard reaction product, [5-(5,6-dihydro-4H-pyridin-3-ylidenemethyl)furan-2-yl]-methanol (F3-A), from an aqueous glucose (Glc) and lysine (Lys) Maillard reaction (MR) model system. Here, we investigate further the mechanisms for anti-inflammatory effects of this product in Caco-2 and RAW 264.7 cells. The anti-inflammatory capacity of F3-A recovered from Glc–Lys MR mixture and a synthesized product were compared with those of the specific inducible nitric oxide synthase (iNOS) inhibitor, aminoguanidine (AG), and the nuclear factor-kappa B (NF-κB) inhibitor, pyrrolidine dithiocarbamate (PDTC). F3-A produced a dose-dependent inhibition of extracellular nitric oxide (NO) production and iNOS translation in Caco-2 cells induced with interferon gamma (IFN-γ) and phorbol 12-myristate 13-acetate (PMA), and these effects were more potent than those obtained with AG. Moreover, a combination of F3-A and AG to attenuate intestinal inflammation was additive. However, F3-A inhibited only intracellular NO production in RAW 264.7 cells and did not inhibit COX-2 or NF-κB in either cell line. We conclude that the anti-inflammatory properties of F3-A are cell specific, working through different mechanism between macrophages and intestinal cells.

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Abbreviations

AG:

Aminoguanidine

COX-2:

Cyclooxygenase-2

DAF-2:

4,5-Diaminofluorescein

DAF-2DA:

4,5-Diaminofluorescein diacetate

DAF-2T:

Triazolofluorescein

DCM:

Dichloromethane

DMEM:

Dulbecco’s modified Eagle’s medium

FBS:

Fetal bovine serum

F3-A:

[5-(5,6-Dihydro-4H-pyridin-3-ylidenemethyl)furan-2-yl]-methanol

F3-AC19:

Isolated F3-A purified by flash chromatography

F3-AS:

Synthesized F3-A

Glc:

Glucose

HMF:

Hydroxymethylfurfural

HMFA:

5-Hydroxymethyl-2-furoic acid

IBD:

Inflammatory bowel disease

IECs:

Intestinal epithelial cells

IFN-γ:

Interferon gamma

IL-6:

Interleukin-6

IL-8:

Interleukin 8

iNOS:

Inducible nitric oxide synthase

LPS:

Lipopolysaccharide

Lys:

Lysine

3-MCP:

3-Methyl-1,2-cyclopentanedione

MEM:

Minimum essential medium

MR:

Maillard reaction

MRPs:

Maillard reaction products

MTT:

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

NF-κB:

Nuclear factor-kappa B

NMR:

Nuclear magnetic resonance

NO:

Nitric oxide

NO2 − :

Nitrite

NO3 − :

Nitrate

NOD:

Nitric oxide dioxygenase

PDTC:

Pyrrolidine dithiocarbamate

PMA:

Phorbol 12-myristate 13-acetate

Rf:

Retention factor

RT-PCR:

Reverse transcription polymerase chain reaction

TLC:

Thin layer chromatography

TBST:

Tris-buffered saline-Tween

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Acknowledgments

This study was supported by a NSERC-discovery grant to DDK. We also thank Yongting Chen for help with protein and mRNA extraction.

Compliance with Ethical Standards

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Food, Nutrition, and Health, Faculty of Land and Food Systems, University of British Columbia, 2205 East Mall, Vancouver, BC, V6T 1Z4, Canada

    Xiu-Min Chen & David D. Kitts

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  1. Xiu-Min Chen
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  2. David D. Kitts
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Correspondence to David D. Kitts.

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Chen, XM., Kitts, D.D. Evidence for inhibition of nitric oxide and inducible nitric oxide synthase in Caco-2 and RAW 264.7 cells by a Maillard reaction product [5-(5,6-dihydro-4H-pyridin-3-ylidenemethyl)furan-2-yl]-methanol. Mol Cell Biochem 406, 205–215 (2015). https://doi.org/10.1007/s11010-015-2438-7

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  • DOI: https://doi.org/10.1007/s11010-015-2438-7

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