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Differential distribution of annexins-I, -II, -IV, and -VI in synovium
Abstract and Figures
To examine the distribution of four annexins in non-inflamed rheumatoid arthritic and osteoarthritic synovial tissue.
Frozen sections were stained with monoclonal antibodies (MAb) specific for annexins-I, -II, -IV, and -VI, and for cell lineage related markers including CD68 and CD14 (macrophages), prolyl hydroxylase (fibroblasts), and CD3 (T cells).
Each of the annexins was present in synovial tissues in significant amounts in the three groups studied. Annexin-I was predominantly found within the synovial lining layer and double labelling showed it to be present predominantly in cells of the macrophage lineage. In rheumatoid specimens there was increased staining within the lining layer, perivascularly and on macrophages within the tissue stroma. Annexin-II was present in a distribution similar to that of annexin-I, but with more prominent perivascular staining. Annexins-IV and -VI were seen chiefly in association with areas of lymphocyte infiltration in rheumatoid tissue, whereas annexins-I and -II were absent from these areas. Endothelial cells stained weakly positive for annexins-I and -II, and more strongly for -IV and -VI.
This study demonstrates that annexins (particularly annexin-I, a putative mediator of the anti-inflammatory activities of glucocorticoids) are abundant in rheumatoid and non-rheumatoid synovial tissue, annexins-IV and -VI having a distribution distinct from that of -I and -II.
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... AnxA1 has been found to be expressed in human RA synovial tissue (56)(57)(58) and has been identified as an important endogenous anti-inflammatory mediator in several animal models of RA (59). ...
... Indeed, several authors described that among the three members of the N-formyl peptide receptor family, FPR2 could mediate anti-inflammatory effects (54), playing a role in the pathogenesis of RA. Moreover, AnxA1, expressed in human RA synovial tissue (56)(57)(58), has been recognized as a significant endogenous antiinflammatory mediator in several animal models of RA. ...
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by a progressive symmetric inflammation of the joints resulting in bone erosion and cartilage destruction with a progressive loss of function and joint deformity. An increased number of findings support the role of innate immunity in RA: many innate immune mechanisms are responsible for producing several cytokines and chemokines involved in RA pathogenesis, such as Tumor Necrosis Factor (TNF)-α, interleukin (IL)-6, and IL-1. Pattern recognition receptors (PRRs) play a crucial role in modulating the activity of the innate arm of the immune response. We focused our attention over the years on the expression and functions of a specific class of PRR, namely formyl peptide receptors (FPRs), which exert a key function in both sustaining and resolving the inflammatory response, depending on the context and/or the agonist. We performed a broad review of the data available in the literature on the role of FPRs and their ligands in RA. Furthermore, we queried a publicly available database collecting data from 90 RA patients with different clinic features to evaluate the possible association between FPRs and clinic-pathologic parameters of RA patients.
... In patients suffering from RA, discrete Anx-1 expression has been observed in monocytes of the synovial membrane [80]. In adjuvant-induced arthritis rat model, the endogenous Anx-1 mediates the anti-inflammatory effect of dexamethasone through the modulation of synovial TNF-a release and neutrophil recruitment [81]. ...
Inflammation is a physiological intrinsic host response to injury meant for removal of noxious stimuli and maintenance of homeostasis. It is a defensive body mechanism that involves immune cells, blood vessels and molecular mediators of inflammation. Glucocorticoids (GCs) are steroidal hormones responsible for regulation of homeostatic and metabolic functions of body. Synthetic GCs are the most useful anti-inflammatory drugs used for the treatment of chronic inflammatory diseases such as asthma, chronic obstructive pulmonary disease (COPD), allergies, multiple sclerosis, tendinitis, lupus, atopic dermatitis, ulcerative colitis, rheumatoid arthritis and osteoarthritis whereas, the long term use of GCs are associated with many side effects. The anti-inflammatory and immunosuppressive (desired) effects of GCs are usually mediated by transrepression mechanism whereas; the metabolic and toxic (undesired) effects are usually manifested by transactivation mechanism. Though GCs are most potent anti-inflammatory and immunosuppressive drugs, the common problem associated with their use is GC resistance. Several research studies are rising to comprehend these mechanisms, which would be helpful in improving the GC resistance in asthma and COPD patients. This review aims to focus on identification of new drug targets in inflammation which will be helpful in the resolution of inflammation. The ample understanding of GC mechanisms of action helps in the development of novel anti-inflammatory drugs for the treatment of inflammatory and autoimmune disease with reduced side effects and minimal toxicity.
... The expression of ANXA1 in subpopulations of T lymphocytes CD4+, CD8+ and Treg were assessed. ANXA1 is known to be constitutively expressed on leukocytes and epithelial cells [17,18,42,43]. Depending on the cell stimulus, ANXA1 expression may be increased endogenously in order to regulate the inflammatory processes [18,44]. ...
Paracrine factors secreted by mesenchymal stem cells (MSCs) reportedly modulate inflammation and reparative processes in damaged tissues and have been explored for knee osteoarthritis (OA) therapy. Although various studies have reported the effects of paracrine factors in knee OA, it is not yet clear which paracrine factors directly affect the regeneration of damaged cartilage and which are secreted under various knee OA conditions. In this study, we cultured MSCs derived from three types of tissues and treated each type with IL-1β and TNF-α or not to obtain conditioned medium. Each conditioned medium was used to analyse the paracrine factors related to cartilage regeneration using liquid chromatography-tandem mass spectrometry. Bone marrow-, adipose tissue-, and synovial membrane-MSCs (all-MSCs) exhibited expression of 93 proteins under normal conditions and 105 proteins under inflammatory conditions. It was confirmed that the types of secreted proteins differed depending on the environmental conditions, and the proteins were validated using ELISA. The results of Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis using a list of proteins secreted by all-MSCs under each condition confirmed that the secreted proteins were closely related to cartilage repair under inflammatory conditions. Protein-protein interaction networks were confirmed to change depending on environmental differences and were found to enhance the secretion of paracrine factors related to cartilage regeneration under inflammatory conditions. In conclusion, our results demonstrated that compared with knee OA conditions, the differential expression proteins may contribute to the regeneration of damaged cartilage. In addition, the detailed information on commonly secreted proteins by all-MSCs provides a comprehensive basis for understanding the potential of paracrine factors to influence tissue repair and regeneration in knee OA.
In inflamed joints, enhanced hyaluronic acid (HA) degradation is closely related to the pathogenesis of rheumatoid arthritis (RA). KIAA1199 has been identified as a hyaladherin that mediates the intracellular degradation of HA, but its extracellular function remains unclear. In this study, we found that the serum and synovial levels of secreted KIAA1199 (sKIAA1199) and low-molecular-weight HA (LMW-HA, MW < 100 kDa) in RA patients were significantly increased, and the positive correlation between them was shown for the first time. Of note, treatment with anti-KIAA1199 mAb effectively alleviated the severity of arthritis and reduced serum LMW-HA levels and cytokine secretion in collagen-induced arthritis (CIA) mice. In vitro, sKIAA1199 was shown to mediate exogenous HA degradation by attaching to the cell membrane of RA fibroblast-like synoviosytes (RA FLS). Furthermore, the HA-degrading activity of sKIAA1199 depended largely on its adhesion to the membrane, which was achieved by its G8 domain binding to ANXA1. In vivo, kiaa1199-KO mice exhibited greater resistance to collagen-induced arthritis. Interestingly, this resistance could be partially reversed by intra-articular injection of vectors encoding full-length KIAA1199 instead of G8-deleted KIAA119 mutant, which further confirmed the indispensable role of G8 domain in KIAA1199 involvement in RA pathological processes. Mechanically, the activation of NF-κB by interleukin-6 (IL-6) through PI3K/Akt signaling is suggested to be the main pathway to induce KIAA1199 expression in RA FLS. In conclusion, our study supported the contribution of sKIAA1199 to RA pathogenesis, providing a new therapeutic target for RA by blocking sKIAA1199-mediated HA degradation.
Aim:
To explore disease-associated molecules in rheumatoid arthritis (RA), we comprehensively analyzed phosphoproteins purified from RA synoviocytes.
Method:
Synoviocytes were obtained from three patients with RA and three patients with osteoarthritis (OA). Profiles of phosphoproteins purified from the synoviocytes were compared by two-dimensional differential gel electrophoresis (2D-DIGE) between the RA and OA groups. Protein spots with significantly different phosphorylation levels were identified by mass spectrometry. Recombinant protein of annexin A4 (ANXA4), one of the identified phosphoproteins, was transfected into synoviocytes from an OA patient to mimic RA synoviocytes and humoral factor secretion was compared between rANXA4-transfected and non-transfected synoviocytes under a tumor necrosis factor-α (TNFα)-stimulated condition.
Results:
In 2D-DIGE, 318 phosphoprotein spots were detected, of which 94 spots showed significantly different intensities between the two groups (P < 0.05). Among the 94 spots, 22 spots showed two-fold or higher intensity and one spot showed less than 1/2-fold intensity in the RA group compared to the OA group. From the 22 spots, 11 phosphoproteins were identified, which included kinases, carrier and chaperone proteins, cytoskeletal proteins, proteases and calcium-binding proteins. One of the identified calcium-binding proteins was ANXA4, an exocytosis-regulating protein. The transfected rANXA4 was found to be phosphorylated intracellularly, and secretion of chemokine (C-X-C motif) ligand 1 and interleukin-8 induced by TNFα stimulation was significantly suppressed by the transfection (P < 0.01).
Conclusion:
The phosphoprotein profile of RA synoviocytes was different from that of OA synoviocytes. This difference would reflect the different pathophysiologies of the diseases. ANXA4 may be one of therapeutic targets in RA.
These anti-inflammatory drugs that are used to treat acute gout are discussed in detail. Their administration, pharmacology, and toxicity are considered. Then, urate-lowering therapy is thoroughly described, again considering the administration, pharmacology, and toxicity of these agents. The widespread mismanagement of gout in general and even specialty medical practice makes this information important for patients and their physicians.
Molecular mediators that are responsible for the acute inflammatory reaction in gout and in other inflammatory processes are reviewed in detail. The acute attack is an example of inflammation mediated by the innate immune system and eventually involves a broad array of mediators. All of this serves as a guide for the therapy of the acute attack of gout.KeywordsSynovial FluidNADPH OxidaseAcute Inflammatory ResponseNeutrophil ChemotaxisAcute GoutThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
Malaria is one of the most widespread and serious parasitic diseases worldwide. Currently available antimalarial drugs have side effects, and many strains of Plasmodia have developed resistance to such drugs. The present review examines the use of annexins (ANXs) and of natural peptides from snake venom as a new class of anti-malarial agents, with the key property of reducing inflammation. Severe cases of malaria manifest elevated serum levels of liver enzymes, inflammation, fibrin deposition, apoptosis, and reduction in peripheral CD8+ T cells. The annexin-A1/5 (ANXA-1/5) proteins trigger inflammation via increased expression of diverse cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin- (IL-10), however, by shielding microbial phospholipids they prevent injury via damage-associated molecular patterns (DAMPs). Here, we also review an in silico -based bioengineering approach that may allow for a better design, synthesis and characterization of novel peptides from snake venom as a more effective approach for treatment due to their improved antimalarial activity.
Aim: To investigate effect of bizhongxiao decoction (BZXD) on changes of protein in synovitis of rats with collagen-induced arthritis (CIA) and analyse mechanism of BZXD on treating rheumatoid arthritis (RA). Methods: The experiment was completed in the Institute of Combined Traditional Chinese Medicine and Western Medicine, Xiangya Hospital of Central South University and Key Laboratory of Tumor Protein of Ministry of Public Health between May 2004 and February 2005. Totally 70 SD rats were divided randomly into 3 groups. Ten rats were regarded as normal group, and other 60 rats were modeled by type II collagen and complete Freunds adjuvant. Successful rat models were selected and divided into BZXD group and model group. BZXD was consisted of 15 kinds of Chinese herbs such as baihuashe shecao 15g, zhonjiefeng 30 g, danshen 15 g, luoshiteng 20 g, gusuibu 15 g, yimi 30 g, etc., and fried with double distilled water for twice with 30 minutes each time. Then, liquid was mixed, filtered with G4 filter and concentrated in water bath at 60°C with 116 g raw materials in each milliliter. Rats in BZXD group were perfused with 10 mL/kg BZXD (equal to 62.1 g raw materials) twice a day; rats in model group were perfused with 10 mL/kg saline twice a day; rats in normal group were drunk water freely. The total proteins of synovial tissue of rats in normal group, model group and BZXD group were separated by two-dimensional gel electrophoresis (2-DE), respectively. Then gets were stained by Coomassie brilliant blue, scanned by Image Scanner. Relative abundance of the protein spots was calculated by comparing spots density volume on the gel. The differentially expressed protein spots were identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching, then some of them were validated by western blotting. Results: Among 70 rats, 7 were lost because of unsuccessful modeling, and totally 63 rats, 26 in BZXD group, 27 in model group and 10 in control group, entered the final analysis. 1 Reproducible 2DE maps of synovial tissue were established in normal group, experimental arthritis model group and BZXD group respectively, the average spots of protein were 947±39, 994±41 and 1 031±52 respectively, and the match rates were 92%, 91% and 94.2%. 2 Totally 55 protein spots were twice than expression of synovial tissue in normal group, experimental arthritis model group and BZXD group. This suggested that the functions of these proteins involved in metabolism, energy generation, transportation, anti-oxidation, signal transduction and protein folding etc. Then the different expressions of annexin 1 and aldolase A in normal group, experimental arthritis model group and BZXD group by western blotting were similar to those of proteomic results. Conclusion: Synovium of joint of rheumtoid arthritis is a complex process which is involved by multiple proteins. Histological analysis of proteins in synovium tissue is one of methods for screening related protein of lesion. Annexin 1 and aldolase A are related to CIA synovitis, and BZXD has a good effect on CIA by regulating protein expression of synovial tissue of rats.
Clinical and experimental observations revealed that glucocorticoid-deficient states are associated with an enhanced inflammatory response. The antiinflammatory response of pharmacological doses of glucocorticoids has been tentatively attributed to the induction of lipocortin-I. To determine whether glucocorticoid deficiency causes lipocortin-I down-regulation, the expression of lipocortin-I mRNA and protein was quantified in rats with and without adrenalectomy (ADX). The mRNA of lipocortin-I was quantified by polymerase chain reaction, using a constant amount of modified lipocortin-I cDNA transcript as an internal standard. The lipocortin-I mRNA was decreased by 56 +/- 14% in lung tissue of ADX rats. This down-regulation of lipocortin-I mRNA was not due to a nonspecific effect of ADX, since the mRNA levels of other proteins (c-fos, c-myc, c-erbA beta, and metallothionein-II) remained unchanged. The decrease in lipocortin-I mRNA in ADX rats was reflected by a corresponding decrease in tissue (lung, spleen, liver, and kidney) lipocortin-I protein content, as assessed by quantitative Western blot analysis. Thus, ADX causes a decline in lipocortin-I message and protein, an observation compatible with the increased susceptibility to inflammatory reactions in glucocorticoid deficiency.
The revised criteria for the classification of rheumatoid arthritis (RA) were formulated from a computerized analysis of 262 contemporary, consecutively studied patients with RA and 262 control subjects with rheumatic diseases other than RA (non-RA). The new criteria are as follows: 1) morning stiffness in and around joints lasting at least 1 hour before maximal improvement; 2) soft tissue swelling (arthritis) of 3 or more joint areas observed by a physician; 3) swelling (arthritis) of the proximal interphalangeal, metacarpophalangeal, or wrist joints; 4) symmetric swelling (arthritis); 5) rheumatoid nodules; 6) the presence of rheumatoid factor; and 7) radiographic erosions and/or periarticular osteopenia in hand and/or wrist joints. Criteria 1 through 4 must have been present for at least 6 weeks. Rheumatoid arthritis is defined by the presence of 4 or more criteria, and no further qualifications (classic, definite, or probable) or list of exclusions are required. In addition, a “classification tree” schema is presented which performs equally as well as the traditional (4 of 7) format. The new criteria demonstrated 91–94% sensitivity and 89% specificity for RA when compared with non-RA rheumatic disease control subjects.
Annexin self-association was studied with 90-degrees light scattering and resonance energy transfer between fluorescein (donor) and eosin (acceptor) labeled proteins. Synexin (annexin VII), p32 (annexin IV), and p67 (annexin VI) self-associated in a Ca2+ -dependent manner in solution. However, this activity was quite labile and, especially for p32 and p67, was not consistently observed. When bound to chromaffin granule membranes, the three proteins consistently self-associated and did so at Ca2+ levels (pCa 5.0-4.5) approximately 10-fold lower than required when in solution. Phospholipid vesicles containing phosphatidylserine and phosphatidylethanolamine (1:1 or 1:3) were less effective at supporting annexin polymerization than were those containing phosphatidylserine and phosphatidylcholine (1:0, 1:1, or 1:3). The annexins bound chromaffin granule membranes in a positively cooperative manner under conditions where annexin self-association was observed, and both phenomena were inhibited by trifluoperazine. Ca2+ -dependent chromaffin granule membrane aggregation, induced by p32 or synexin, was associated with intermembrane annexin polymerization at Ca2+ levels less than pCa 4, but not at higher Ca2+ concentrations, suggesting that annexin self-association may be necessary for membrane contact at low Ca2+ levels but not at higher Ca2+ levels where the protein may bind two membranes as a monomer.
The mechanisms that regulate inflammatory responses, and their dysfunction in chronic inflammatory diseases, are poorly understood. Lipocortin 1 is a potential mediator of the anti-inflammatory effects of glucocorticoid hormones. Following the discovery of putative receptors on phagocytes for lipocortin 1, Nick Goulding and Paul Guyre here offer one explanation for the self-limitation of inflammation and the impairment of the mechanism in chronic inflammatory disease.
Autoantibodies to the antiinflammatory protein lipocortin-1 have been found in patients with rheumatoid arthritis (RA) receiving oral glucocorticoids. The highest antibody titers correlated with a requirement for high maintenance doses of steroid (> 7.5 mg/day prednisolone). Forty-two patients with RA were grouped according to high or low autoantibody titer. In 18 patients, peripheral blood leukocyte counts and phenotypic analysis were performed before and 4 h after a single intravenous (iv) dose of 100 mg hydrocortisone. The group with low titer antibody exhibited a normal poststeroid peripheral blood lymphopenia, but the response in the group with high antibody titer was considerably blunted. In a 2nd study, 24 patients received 3 separate doses of 1000 mg iv methylprednisolone. After 8 weeks the group with the high titer antibody had shown no improvements in clinical or laboratory variables observed in the group with low titer antibody. Thus, the presence of high titer antilipocortin-1 antibody is associated with impaired responses to glucocorticoid therapy both in terms of clinical efficacy and effects on the immune system. This could explain the relative glucocorticoid resistance reported in a proportion of patients with RA.
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