formazans and Arthritis--Rheumatoid

To investigate whether protease-activated receptor 1 (PAR-1) and/or PAR-2 promotes the invasiveness/proliferation of synovial fibroblasts (SFs) and to determine the signaling mechanisms of these pathways.. SFs were isolated from the synovial tissue of patients with rheumatoid arthritis (RA), patients with osteoarthritis (OA), and PAR-1- or PAR-2-knockout (KO) mice. Expression of PAR-1 and PAR-2 was detected by immunofluorescence and Western blotting. The invasion and proliferation of SFs were measured by invasion assay and MTT assay, respectively. Matrix metalloproteinase 2 (MMP-2) and MMP-9 were detected by zymography, and cytokines were measured by enzyme-linked immunosorbent assay.. PAR-1 and PAR-2 were colocalized with SFs in RA and OA synovium and, to a considerably lesser extent, in normal synovium. Inhibition of PAR-2 by small interfering RNA (siRNA) inhibited RASF invasion and proliferation, whereas blocking of PAR-1 by siRNA had the reverse effects. SFs from PAR-2-KO mice exhibited slower rates of proliferation and invasion. SFs from PAR-1-KO mice produced less MMP-2 and, in response to tumor necrosis factor α (TNFα) stimulation, had increased MMP-9 secretion when compared to SFs from wild-type and PAR-2-KO mice. Inhibition of PAR-1, but not PAR-2, stimulated the secretion of interleukin-17 (IL-17) and TNFα by RASFs. Furthermore, PAR-1 and PAR-2 had opposing effects on the activation of ERK, p38, and NF-κB.. Activation of PAR-1 stimulates MMP-2 secretion, inhibits RASF growth and invasion, and decreases production of IL-17 and TNFα by RASFs, whereas activation of PAR-2 stimulates RASF growth and invasion and increases production of TNFα. Thus, although PAR-1 and PAR-2 are coexpressed by RASFs, PAR-2 alone appears to be responsible for the aggressive properties of RASFs and is likely to contribute to the pathologic progression of RA.

Topics: Aged; Animals; Arthritis, Rheumatoid; Cell Movement; Cell Proliferation; Cell Survival; Female; Fibroblasts; Formazans; Humans; Male; Matrix Metalloproteinase 9; Mice; Mice, Inbred C57BL; Mice, Knockout; Osteoarthritis; Osteoarthritis, Knee; Receptor, PAR-1; Receptor, PAR-2; RNA Interference; RNA, Small Interfering; Synovial Membrane; Tetrazolium Salts; Transfection; Tumor Necrosis Factor-alpha

Estrogen plays an important modulatory role in the immune system, and is concerned with the pathophysiology of autoimmune diseases such as rheumatoid arthritis (RA), although the mechanism has not yet been clarified. Oral tolerance, a form of specific peripheral tolerance, which is recognized as a new therapeutic strategy, is related to the function of gut-associated lymphoid tissue.. In this study, using collagen-induced arthritis as an animal model of RA, the effects of 17beta-estradiol (E2) on oral tolerance induction were investigated. For induction of oral tolerance, mice were fed 60 microg type II collagen (CII) for 10 consecutive days prior to each CII immunization. Mice in the E2 treatment groups were injected with 5 microg (low dose) or 500 microg (high dose) E2 three times during the induction of oral tolerance.. Oral tolerance induction suppressed the occurrence of arthritis, the proliferative response of splenocytes to CII and the specific DTH response. However, E2 treatment abrogated the suppression, which might be connected with a change in function of Peyer's patch (PP) lymphocytes.. These results suggest that oral tolerance induction might be affected by estrogen treatment through alteration of intestinal immune responses.

Topics: Animals; Apoptosis; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Division; Estradiol; Formazans; Hypersensitivity, Delayed; Immune Tolerance; Immunoglobulin G; Immunohistochemistry; In Situ Nick-End Labeling; Lymphocytes; Male; Mice; Mice, Inbred DBA; Peyer's Patches; Tetrazolium Salts