Proresolving and cartilage-protective actions of resolvin D1 in inflammatory arthritis
Lucy V. Norling, Sarah E. Headland, Jesmond Dalli, Hildur H. Arnardottir, Oliver Haworth, Hefin R. Jones, Daniel Irimia, Charles N. Serhan, Mauro Perretti
Lucy V. Norling, Sarah E. Headland, Jesmond Dalli, Hildur H. Arnardottir, Oliver Haworth, Hefin R. Jones, Daniel Irimia, Charles N. Serhan, Mauro Perretti
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Research Article Immunology Inflammation

Proresolving and cartilage-protective actions of resolvin D1 in inflammatory arthritis

Abstract

Rheumatoid arthritis (RA) is a debilitating disease characterized by persistent accumulation of leukocytes within the articular cavity and synovial tissue. Metabololipidomic profiling of arthritic joints from omega-3 supplemented mice identified elevated levels of specialized proresolving lipid mediators (SPM) including resolvin D1 (RvD1). Profiling of human RA synovial fluid revealed physiological levels of RvD1, which — once applied to human neutrophils — attenuated chemotaxis. These results prompted analyses of the antiarthritic properties of RvD1 in a model of murine inflammatory arthritis. The stable epimer 17R-RvD1 (100 ng/day) significantly attenuated arthritis severity, cachexia, hind-paw edema, and paw leukocyte infiltration and shortened the remission interval. Metabololipidomic profiling in arthritic joints revealed 17R-RvD1 significantly reduced PGE2 biosynthesis, while increasing levels of protective SPM. Molecular analyses indicated that 17R-RvD1 enhanced expression of genes associated with cartilage matrix synthesis, and direct intraarticular treatment induced chondroprotection. Joint protective actions of 17R-RvD1 were abolished in RvD1 receptor–deficient mice termed ALX/fpr2/3–/–. These investigations open new therapeutic avenues for inflammatory joint diseases, providing mechanistic substance for the benefits of omega-3 supplementation in RA.

Authors

Lucy V. Norling, Sarah E. Headland, Jesmond Dalli, Hildur H. Arnardottir, Oliver Haworth, Hefin R. Jones, Daniel Irimia, Charles N. Serhan, Mauro Perretti

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Figure 6

Intraarticular treatment with 17R-RvD1 protects from cartilage degradation during inflammatory arthritis.

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Intraarticular treatment with 17R-RvD1 protects from cartilage degradati...
(A) Arthritis was induced with arthritogenic serum (100 μl, days 0 and 2), mice were treated daily with vehicle (0.1% EtOH) or 17R-RvD1 (100 ng, i.p.), and paws were collected for gene expression analysis on day 8 (see Methods); n = 8–9 mice per group, *P < 0.05 with Mann-Whitney t test. Col2a1, collagen type II α 1; Acan, aggrecan. (B and C) Mice received arthritogenic serum (100 μl, i.p. days 0 and 2) and were treated locally on day 3 with vehicle (left knee; 5 μl PBS containing 0.1% EtOH) or 17R-RvD1 (right knee; 5 μl, 100 ng 17R-RvD1). On day 5, knee joints were collected and stained with toluidine blue; n = 7 mice per group. Naive mice were administered vehicle or 17R-RvD1 locally, and knees collected after 2 days; n = 6 mice per group. (B) Representative images (×20 magnification) of histological sections from naive and arthritic joints are shown. C, cartilage; m, meniscus. Loss of glycosaminoglycans indicated by arrow heads. (C) Cartilage integrity calculated from percentage area of cartilage positive for toluidine blue staining; **P < 0.01 with 2-tailed paired Student’s t test. (D and E) In vitro analyses of chondroprotection utilizing human C28/I2 micromasses. (D) Micromasses were treated with or without IL-1β (30 ng/ml) alone or with 17R-RvD1 (0.1–100 nM) and ECM accumulation evaluated (see Methods); n = 6–10 per group, **P < 0.01, ***P < 0.001 vs. IL-1β with 2-way ANOVA and Bonferroni post-test. Inset representative micromasses stained with Alcian blue prior to dye extraction. (E) FPR2/ALX receptor antagonist WRW4 (10 μM) was added to micromasses 10 minutes prior to 17R-RvD1 and ECM accumulation evaluated after 24 hours; n = 3–7 per group, ***P < 0.001 vs. vehicle, ###P < 0.001 vs. respective control with 2-way ANOVA and Bonferroni post-test. (F) Dependency of Fpr2/3 (ALX) receptor for 17R-RvD1 protection from inflammatory arthritis. Arthritis was induced in Fpr2/3-null (ALX-null) mice (see Methods), mice were treated daily with vehicle (PBS with 0.1% EtOH) or 17R-RvD1 (100 ng, i.p.), and arthritic score was assessed; n = 9–10 mice per group.