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Anti-inflammatory effects of N-acylethanolamines in rheumatoid arthritis synovial cells are mediated by TRPV1 and TRPA1 in a COX-2 dependent manner.

Lowin T, Apitz M, Anders S, Straub RH - Arthritis Res. Ther. (2015)

Bottom Line: The effects of OEA and PEA on SFs were diminished by FAAH inhibition.N-acylethanolamines exert anti-inflammatory effects in SFs.A dual FAAH/COX-2 inhibitor, increasing N-acylethanolamine levels with concomitant TRP channel desensitization, might be a good candidate to inhibit the production of proinflammatory mediators of synovial cells and to reduce erosions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Experimental Rheumatology and Neuroendocrine Immunology, Department of Internal Medicine I, University Hospital Regensburg, Franz Josef Strauss Allee 11, 93042, Regensburg, Germany. torsten.lowin@ukr.de.

ABSTRACT

Introduction: The endocannabinoid system modulates function of immune cells and mesenchymal cells such as fibroblasts, which contribute to cartilage destruction in rheumatoid arthritis (RA). The aim of the study was to determine the influence of N-acylethanolamines anandamide (AEA), palmitoylethanolamine (PEA) and oleylethanolamine (OEA) on several features of arthritic inflammation in vitro (human material) and in vivo (a mouse model).

Methods: Immunofluorescence and western blotting were used to detect cannabinoid receptors and related enzymes. Cytokines and MMP-3 were measured by ELISA. Intracellular signaling proteins were detected by proteome profiling. Proliferation was quantified by CTB reagent. Adhesion was assessed by the xCELLigence system. After onset of collagen type II arthritis, mice were treated daily with the FAAH inhibitor JNJ1661010 (20 mg/kg) or vehicle.

Results: IL-6, IL-8 and MMP-3 (determined only in synovial fibroblasts (SFs)) were downregulated in primary synoviocytes and SFs of RA and OA after AEA, PEA and OEA treatment. In SFs, this was due to activation of TRPV1 and TRPA1 in a COX-2-dependent fashion. FAAH inhibition increased the efficacy of AEA in primary synoviocytes but not in SFs. The effects of OEA and PEA on SFs were diminished by FAAH inhibition. Adhesion to fibronectin was increased in a CB1-dependent manner by AEA in OASFs. Furthermore, elevation of endocannabinoids ameliorated collagen-induced arthritis in mice.

Conclusions: N-acylethanolamines exert anti-inflammatory effects in SFs. A dual FAAH/COX-2 inhibitor, increasing N-acylethanolamine levels with concomitant TRP channel desensitization, might be a good candidate to inhibit the production of proinflammatory mediators of synovial cells and to reduce erosions.

No MeSH data available.


Related in: MedlinePlus

Influence of anandamide (AEA) with or without fatty acid amide hydrolase (FAAH) inhibition. Effects are described on IL-6 (a, b), IL-8 (c, d) and TNF (e, f) production in RA mixed synovial cell cultures. Cytokine production was determined under normoxic (20 % O2) conditions. *p <0.01 vs. control (100 %). Mann-Whitney U test was used for comparisons between groups; paired t test was used for comparisons vs. control. All data are given as box and vertical dot blots. The boundary of the box closest to zero indicates the 25th percentile, the line within each box indicates the median and the upper boundary of the box the 75th percentile. Error bars below and above the box indicate the 10th and 90th percentiles. Blue color represents treatment with the FAAH inhibitor JNJ1661010 (1 μM). IL-6 interleukin-6, IL-8 interleukin-8, RA rheumatoid arthritis, TNF tumor necrosis factor
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Fig1: Influence of anandamide (AEA) with or without fatty acid amide hydrolase (FAAH) inhibition. Effects are described on IL-6 (a, b), IL-8 (c, d) and TNF (e, f) production in RA mixed synovial cell cultures. Cytokine production was determined under normoxic (20 % O2) conditions. *p <0.01 vs. control (100 %). Mann-Whitney U test was used for comparisons between groups; paired t test was used for comparisons vs. control. All data are given as box and vertical dot blots. The boundary of the box closest to zero indicates the 25th percentile, the line within each box indicates the median and the upper boundary of the box the 75th percentile. Error bars below and above the box indicate the 10th and 90th percentiles. Blue color represents treatment with the FAAH inhibitor JNJ1661010 (1 μM). IL-6 interleukin-6, IL-8 interleukin-8, RA rheumatoid arthritis, TNF tumor necrosis factor

Mentions: Under these conditions, AEA (at 10−6 M and 10−8 M) and concomitant FAAH inhibition with JNJ1661010 (1 μM) reduced the production of IL-6 and IL-8 by RA but not OA mixed synoviocytes (Fig. 1b, d). TNF production was augmented only by OA synoviocytes (at 10−8 M without FAAH inhibition and 10−10 M with FAAH inhibition) (Fig. 1e). Average control values for cytokines (dotted line at 100 % in Fig. 1) were 110 ± 44 ng/ml (OA) and 148 ± 49 ng/ml (RA) for IL-6, 103 ± 49 ng/ml (OA) and 171 ± 56 ng/ml (RA) for IL-8, and 66 ± 68 pg/ml (OA) and 163 ± 166 pg/ml (RA) for TNF.Fig. 1


Anti-inflammatory effects of N-acylethanolamines in rheumatoid arthritis synovial cells are mediated by TRPV1 and TRPA1 in a COX-2 dependent manner.

Lowin T, Apitz M, Anders S, Straub RH - Arthritis Res. Ther. (2015)

Influence of anandamide (AEA) with or without fatty acid amide hydrolase (FAAH) inhibition. Effects are described on IL-6 (a, b), IL-8 (c, d) and TNF (e, f) production in RA mixed synovial cell cultures. Cytokine production was determined under normoxic (20 % O2) conditions. *p <0.01 vs. control (100 %). Mann-Whitney U test was used for comparisons between groups; paired t test was used for comparisons vs. control. All data are given as box and vertical dot blots. The boundary of the box closest to zero indicates the 25th percentile, the line within each box indicates the median and the upper boundary of the box the 75th percentile. Error bars below and above the box indicate the 10th and 90th percentiles. Blue color represents treatment with the FAAH inhibitor JNJ1661010 (1 μM). IL-6 interleukin-6, IL-8 interleukin-8, RA rheumatoid arthritis, TNF tumor necrosis factor
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4644337&req=5

Fig1: Influence of anandamide (AEA) with or without fatty acid amide hydrolase (FAAH) inhibition. Effects are described on IL-6 (a, b), IL-8 (c, d) and TNF (e, f) production in RA mixed synovial cell cultures. Cytokine production was determined under normoxic (20 % O2) conditions. *p <0.01 vs. control (100 %). Mann-Whitney U test was used for comparisons between groups; paired t test was used for comparisons vs. control. All data are given as box and vertical dot blots. The boundary of the box closest to zero indicates the 25th percentile, the line within each box indicates the median and the upper boundary of the box the 75th percentile. Error bars below and above the box indicate the 10th and 90th percentiles. Blue color represents treatment with the FAAH inhibitor JNJ1661010 (1 μM). IL-6 interleukin-6, IL-8 interleukin-8, RA rheumatoid arthritis, TNF tumor necrosis factor
Mentions: Under these conditions, AEA (at 10−6 M and 10−8 M) and concomitant FAAH inhibition with JNJ1661010 (1 μM) reduced the production of IL-6 and IL-8 by RA but not OA mixed synoviocytes (Fig. 1b, d). TNF production was augmented only by OA synoviocytes (at 10−8 M without FAAH inhibition and 10−10 M with FAAH inhibition) (Fig. 1e). Average control values for cytokines (dotted line at 100 % in Fig. 1) were 110 ± 44 ng/ml (OA) and 148 ± 49 ng/ml (RA) for IL-6, 103 ± 49 ng/ml (OA) and 171 ± 56 ng/ml (RA) for IL-8, and 66 ± 68 pg/ml (OA) and 163 ± 166 pg/ml (RA) for TNF.Fig. 1

Bottom Line: The effects of OEA and PEA on SFs were diminished by FAAH inhibition.N-acylethanolamines exert anti-inflammatory effects in SFs.A dual FAAH/COX-2 inhibitor, increasing N-acylethanolamine levels with concomitant TRP channel desensitization, might be a good candidate to inhibit the production of proinflammatory mediators of synovial cells and to reduce erosions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Experimental Rheumatology and Neuroendocrine Immunology, Department of Internal Medicine I, University Hospital Regensburg, Franz Josef Strauss Allee 11, 93042, Regensburg, Germany. torsten.lowin@ukr.de.

ABSTRACT

Introduction: The endocannabinoid system modulates function of immune cells and mesenchymal cells such as fibroblasts, which contribute to cartilage destruction in rheumatoid arthritis (RA). The aim of the study was to determine the influence of N-acylethanolamines anandamide (AEA), palmitoylethanolamine (PEA) and oleylethanolamine (OEA) on several features of arthritic inflammation in vitro (human material) and in vivo (a mouse model).

Methods: Immunofluorescence and western blotting were used to detect cannabinoid receptors and related enzymes. Cytokines and MMP-3 were measured by ELISA. Intracellular signaling proteins were detected by proteome profiling. Proliferation was quantified by CTB reagent. Adhesion was assessed by the xCELLigence system. After onset of collagen type II arthritis, mice were treated daily with the FAAH inhibitor JNJ1661010 (20 mg/kg) or vehicle.

Results: IL-6, IL-8 and MMP-3 (determined only in synovial fibroblasts (SFs)) were downregulated in primary synoviocytes and SFs of RA and OA after AEA, PEA and OEA treatment. In SFs, this was due to activation of TRPV1 and TRPA1 in a COX-2-dependent fashion. FAAH inhibition increased the efficacy of AEA in primary synoviocytes but not in SFs. The effects of OEA and PEA on SFs were diminished by FAAH inhibition. Adhesion to fibronectin was increased in a CB1-dependent manner by AEA in OASFs. Furthermore, elevation of endocannabinoids ameliorated collagen-induced arthritis in mice.

Conclusions: N-acylethanolamines exert anti-inflammatory effects in SFs. A dual FAAH/COX-2 inhibitor, increasing N-acylethanolamine levels with concomitant TRP channel desensitization, might be a good candidate to inhibit the production of proinflammatory mediators of synovial cells and to reduce erosions.

No MeSH data available.


Related in: MedlinePlus