Resveratrol displays anti-inflammatory properties in an ex vivo model of immune mediated inflammatory arthritis

Abstract

Background: Resveratrol is a natural polyphenol found in berries, roots and wine that is well known to have anti-inflammatory and anti-oxidative properties. The anti-inflammatory effect has been reported for both immune cells and connective tissues, but only few studies have investigated effects on immune mediated inflammatory arthritis. None of which have studied this effect when combining resveratrol with methotrexate or adalimumab, two major drugs in the treatment of immune mediated inflammatory arthritis.We therefore aimed to investigate the anti-inflammatory effect of resveratrol alone and in combination with methotrexate or adalimumab in ex vivo models of immune mediated inflammatory arthritis. We furthermore aimed to describe any variations in this effect based on disease activity and cellular composition of the synovial fluid infiltrate.

Methods: Synovial fluid mononuclear cells from patients with rheumatoid arthritis (n = 7) and spondyloarthritis (n = 7) were cultured for either 48 h or 21 days. In both models, synovial fluid mononuclear cells were treated with resveratrol alone or in combination with methotrexate or adalimumab. Monocyte chemoattractant protein 1, matrix metalloproteinase 3 and tartrate resistant acidic phosphatase were measured to quantify inflammation, enzymatic degradation and osteoclast differentiation, respectively.

Results: Resveratrol reduced monocyte chemoattractant protein 1 production by synovial fluid mononuclear cells significantly (p = 0.005) compared to untreated controls. The effect of resveratrol was greatest in cultures from patients with low disease activity, i.e. DAS28CRP ≤ 3.2 (p = 0.022), and in cultures dominated by lymphocytes (p = 0.03). Further, the combination of methotrexate and resveratrol significantly reduced monocyte chemoattractant protein 1 levels compared with methotrexate alone in cultures from patients with low disease activity (p = 0.016), and in cultures with high lymphocyte count (p = 0.011). Resveratrol did not significantly affect matrix metalloproteinase 3 and tartrate resistant acidic phosphatase production.

Conclusion: Resveratrol has anti-inflammatory properties in our ex vivo model of immune mediated inflammatory arthritis. Results show an additive effect of resveratrol, when combined with methotrexate in samples dominated by lymphocytes and samples from patients with low disease activity. This suggests further investigations in vitro and whether this effect may also be present in a clinical setting.

Keywords: Anti-inflammatory agents; Low disease activity; MCP-1; Methotrexate; Monocyte chemoattractant protein 1; Resveratrol; Rheumatoid arthritis; Spondyloarthritis.

Fig. 1

Cell culture viability assessed in microscope (× 10) of 48 h- and 21 days SFMC cultures after incubation period. No immediate cell loss after RSV treatment (25 μM) compared to untreated (UT) cultures. 21 days SFMC cultures showed forming of multinucleated cells

Fig. 2

Concentrations of MCP-1 (a, n = 14 and b, n = 8), MMP3 (c, n = 9) and TRAP activity (d, n = 8) in untreated (UT) and resveratrol (RSV) treated cultures measured by ELISA and enzyme activity assay in their respective supernatants. UT is presented as a negative control to its in-patient corresponding RSV treated culture, * p < 0.05. RSV concentration was 25 μM

Fig. 3

Ratio of MCP-1 in resveratrol (RSV) treated cultures (n = 14) divided in groups of patients with rheumatoid arthritis (RA) and spondyloarthritis (SpA). MCP-1 was measured by ELISA. RSV concentration was 25 μM

Fig. 4

Ratio of MCP-1 in resveratrol (RSV) treated cultures (n = 11) plotted against DAS28CRP (a), and the percentage of monocytes (b) and lymphocytes (c) in synovial fluid of the donor. d-f depicts group comparisons as median and interquartiles. *p < 0.05 vs. negative control. MCP-1 was measured by ELISA. RSV concentration was 25 μM

Fig. 5

Ratios of MCP-1 (a, n = 14 and b, n = 8 (RSV and MTX + RSV) or n = 7 (MTX, ADA and ADA + RSV)) and MMP3 (c, n = 9) concentrations, and TRAP activity (d, n = 8) in supernatants from cultures with the noted substance or a combination of resveratrol (RSV), Ebetrex (MTX) and Adalimumab (ADA). Concentrations were: RSV; 25 μM, MTX; 0.5 μg/ml, Adalimumab; 5 μg/ml. MCP-1 and MMP3 were measured by ELISA and TRAP with an enzyme activity assay. Data is presented as a dot for each measurement; box and bars represent median and interquartile range. *p < 0.05 vs. negative control

Fig. 6

MCP-1 ratios as median and interquartiles in cultures treated with ebetrex (MTX) alone or in combination with resveratrol (RSV). Group comparisons were based on a) Disease Activity Score 28 (DAS28CRP), b) the percentage of monocytes and c) the percentage of lymphocytes in synovial fluid of the donor. *p < 0.05 vs. negative control. MCP-1 was measured by ELISA. Concentrations were: RSV = 25 μM and MTX = 0.5 μg/ml

Fig. 7

MCP-1 ratios as median and interquartiles in cultures treated with adalimumab (ADA) alone or in combination with resveratrol (RSV). Group comparisons were based on a) Disease Activity Score 28 (DAS28CRP), b) the percentage of monocytes and c) the percentage of lymphocytes in synovial fluid of the donor. *p < 0.05 vs. negative control. MCP-1 was measured by ELISA. Concentrations were: RSV = 25 μM and ADA = 5 μg/ml