Ann Dermatol.  2018 Oct;30(5):566-574. 10.5021/ad.2018.30.5.566.

The Suppressive Effect of Butyrate and Bromopyruvate on Inflammatory Cytokine Production and Short Chain Fatty Acid Receptor Expression by Blood Mononuclear Cells in Patients with Behçet's Disease

• Yun SJ
• Kim K
• Lee ES
• Park S

• KMID: 2419747
• DOI: http://doi.org/10.5021/ad.2018.30.5.566

Abstract

BACKGROUND
Controlling inflammation is a therapeutic goal of various autoimmune/autoinflammatory diseases including Behçet's disease (BD). The immunomodulatory effect of metabolites or metabolic analogs such as butyrate and 3-bromopyruvate has been observed in animal disease models.
OBJECTIVE
We attempted to evaluate the effect of butyrate and 3-bromopyruvate on the inflammatory cytokine production by peripheral blood mononuclear cells (PBMCs) isolated from patients with mucocutaneous involvement of BD.
METHODS
PBMCs isolated from 11 patients with BD and 10 healthy controls were stimulated with lipopolysaccharide in the presence of butyrate or 3-bromopyruvate. Butyrate receptor and cytokine messenger ribonucleic acid (mRNA) expression was analyzed by real-time reverse transcription polymerase chain reaction. Cytokine secretion was assessed by enzyme-linked immunosorbent assay. PBMCs survival was analyzed by flow cytometry.
RESULTS
Bromopyruvate or butyrate treatment suppressed inflammatory cytokine production in PBMCs from all our subjects. Bromopyruvate also reduced PBMCs survival while butyrate did not. As the effect of butyrate was slightly greater in BD patients than in healthy controls, we analyzed butyrate receptor expression and found that lipopolysaccharide-induced free fatty acid receptor 2 mRNA level in PBMCs was higher in BD patients than in controls.
CONCLUSION
We propose bromopyruvate and butyrate as supplementary therapeutic candidates to control inflammation in patients with BD.

Keyword

Autoimmune diseases; Butyrates; Glycolysis; Inflammation

MeSH Terms

Autoimmune Diseases
Butyrates*
Disease Models, Animal
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Glycolysis
Humans
Inflammation
Polymerase Chain Reaction
Reverse Transcription
RNA
RNA, Messenger
Butyrates
RNA
RNA, Messenger

Figure

• 

  Fig. 1 The effect of bromopyruvate and butyrate on the cytokine mRNA expression and the survival of THP-1 cells. Cytokine mRNA levels in lipopolysaccharide (LPS)-stimulated THP-1 cells in the presence or absence of bromopyruvate (BrPA) or butyrate (Buty) were analyzed by reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Relative mRNA level to the mRNA level at the beginning of culture (A). The percent mRNA level against the mRNA amount in the absence of a drug (B~E). Cell survival (F). Data are presented as median with range of 6 independent experiments with duplication or triplication. IL: interleukin, mRNA: messenger ribonucleic acid, TNF: tumor necrosis factor. *p<0.05, **p<0.01, ***p<0.001.

• 

  Fig. 2 The effect of bromopyruvate and butyrate on the cytokine expression and the survival of peripheral blood mononuclear cells (PBMCs). PBMCs of patients with Behçet's disease (BD) and healthy controls (HC) were stimulated with lipopolysaccharide (LPS) in the presence or absence of bromopyruvate (BrPA) or butyrate (Buty). The indicated cytokine mRNA (A, C, E) and protein levels (B, D, F) were determined by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Enzyme-linked immunosorbent assay, respectively. The relative mRNA levels in the LPS-stimulated PBMCs against the average mRNA level in unstimulated HC PBMCs (A). The percent mRNA level in the LPS/BrPA or Buty - treated PBMCs against the mRNA level in the LPS-stimulated PBMCs (C, E). The percent protein level in the LPS/BrPA or Buty - treated PBMCs against the protein level in the LPS-stimulated PBMCs (D, F). Cell survival was analyzed by flow cytometry (G). Data are presented as median with range. Data from more than 5 independent experiments with duplication or triplication. IL: interleukin, mRNA: messenger ribonucleic acid, TNF: tumor necrosis factor, Inac. BD: patients with inactive BD, Ac. BD: patients with active BD. *p<0.05, **p<0.01, ***p<0.001.

• 

  Fig. 3 Free fatty acid receptor 2 (FFAR2) and FFAR3 mRNA expression in peripheral blood mononuclear cells (PBMCs). The relative mRNA level in ex-vivo PBMCs of each subject against the average mRNA amount of healthy controls (HC) (A). The relative mRNA levels in the lipopolysaccharide (LPS)-stimulated PBMCs against the average mRNA level in unstimulated HC PBMCs (B). The percent mRNA level in the LPS/bromopyruvate (BrPA) or butyrate - treated PBMCs against the mRNA level in the LPS-stimulated PBMCs (C, D). Data are presented as median with range of more than 5 independent experiments with duplication or triplication. mRNA: messenger ribonucleic acid, Inac. BD: patients with inactive BD, Ac. BD: patients with active BD. *p<0.05, **p<0.01, ***p<0.001.

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