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This article is part of the supplement: 25th European Workshop for Rheumatology Research

Poster presentation

Proinflammatory role for AT1 receptors in the rat synovium

AG Price1, JC Lockhart1, JS McLean1 and WR Ferrell2

Author Affiliations

1 Biological Sciences, University of Paisley, UK

2 Centre for Rheumatic Diseases, Royal Infirmary, Glasgow, UK

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Arthritis Research & Therapy 2005, 7(Suppl 1):P74  doi:10.1186/ar1595


The electronic version of this article is the complete one and can be found online at:


Received:11 January 2005
Published:17 February 2005

© 2005 BioMed Central Ltd

Poster presentation

The increasing recognition that angiotensin II (Ang II) has proinflammatory roles suggests it may therefore contribute to chronic inflammatory conditions such as rheumatoid arthritis (RA), but this requires investigation. Walsh and colleagues demonstrated the presence and upregulation of AT1 receptors [1] and angiotensin-converting enzyme [2] in the synovium of RA patients. The aim of this study was to elucidate the role of Ang II in rat knee joint pathophysiology and to investigate the anti-inflammatory potential of AT1 receptor antagonist losartan in models of arthritis. The second phase of this study was to determine the mechanism of any anti-inflammatory action of losartan. In vivo losartan has been reported to undergo hepatic first-pass metabolism [3], yielding two metabolites, EXP-3174 (the active metabolite) and EXP-3179. The latter is structurally similar to the non-selective cyclooxygenase inhibitor indomethacin. Since the anti-inflammatory benefit of cyclooxygenase inhibitors is well documented, we aimed to determine whether any anti-inflammatory benefit associated with losartan was attributable to AT1 receptor antagonism or inhibitory action on the prostanoid system.

In vivo experiments

Experimental arthritis was induced under anaesthesia (halothane/O2/N2O) by intra-articular and peri-articular injection of Freund's complete adjuvant in adult male Wistar rats (350 ± 20 g body weight, mean ± SEM, n = 6). Knee joint swelling was measured using callipers and expressed as the percentage change from pre-induction values. The anti-inflammatory potential of losartan (Merck & Co., USA) was tested both prophylactically and therapeutically. Prophylactically, losartan (15 mg/kg; subcutaneous) was given 1 hour prior to induction of inflammation and every 48 hours thereafter. Therapeutically, 12 days after the induction of inflammation, losartan (15 mg/kg; subcutaneous) treatment was started and dosing continued every 48 hours. The mechanism of action of losartan was investigated using EXP 3174 (Merck & Co.), given prophylactically (15 mg/kg; subcutaneous, maintenance every 48 hours) 1 hour prior to induction of inflammation (Freund's complete adjuvant).

In vitro experiments

U937 human monocytes at a logarithmic phase of growth were harvested and adjusted to 106 cells/ml in the presence of 10 nM phorbol 12-myristate 13-acetate for 3 days to differentiate into adherent macrophages. Cells were then quiesced for 24 hours before experiments commenced. Cells were then stimulated with lipopolysaccharide (50 ng/ml) incubated in the presence of medium (control), Ang II (10-9 mol) or losartan (10-9 mol to 10-6 mol). Supernatants were collected and tumour necrosis factor alpha (TNF-α) levels measured by ELISA.

Prophylactic administration of losartan resulted in a significant reduction in knee joint swelling over 14 days (two-way analysis of variance [ANOVA], P < 0.00001; n = 6). Therapeutic losartan treatment reduced established knee joint swelling compared with vehicle control (two-way ANOVA, P < 0.00001; n = 7). Prophylactic administration of EXP-3174 also resulted in a reduced percentage increase in knee joint diameter, again found to be significant (two-way ANOVA, P < 0.00001; n = 6–8). Losartan inhibited TNF-α production from U937 cells in a dose-dependent manner (cf. LPS-treated cells; n = 3).

The anti-inflammatory benefit of both prophylactic and therapeutic losartan treatment suggests both an important role for Ang II in arthritis, and a novel therapeutic application for AT1 receptor antagonists. This anti-inflammatory action may be due to inhibition of TNF-α release from macrophages.

Acknowledgements

This study was funded by University of Paisley Research Funds and Merck & Co, USA.

References

  1. Walsh DA, Suzuki T, Knock GA, Blake DR, Polak JM, Wharton J: AT1 receptor characteristics of angiotensin analogue binding in human synovium.

    Br J Pharmacol 1994, 112:435-442. PubMed Abstract OpenURL

  2. Walsh DA, Mapp PI, Wharton J, Polak JM, Blake DR: Neuropeptide degrading enzymes in normal and inflamed human synovium.

    Am J Pathol 1993, 142:1610-1621. PubMed Abstract OpenURL

  3. Krämer C, Sunkomat J, Witte J, Luchtefeld M, Walden M, Schmidt B, Böger RH, Forssmann W-G, Drexler H, Schieffer B: Angiotensin II receptor-independent antiinflammatory and antiaggregatory properties of losartan: role of the active metabolite EXP3179.

    Circ Res 2002, 90:770-776. PubMed Abstract | Publisher Full Text OpenURL