Paeoniflorin-6'-O-benzene sulfonate ameliorates progression of adjuvant-induced arthritis by inhibiting interaction between Ahr and GRK2 of fibroblast-like synoviocytes / 中国药理学与毒理学杂志

OBJECTIVE Aryl hydrocarbon receptor (Ahr) is thought to be a crucial factor that regulates immune responses, which may be involved in the pathogenesis of autoimmune inflammation including rheumatoid arthritis (RA). The results of our group in recent years have shown that CP-25, a novel ester derivative of paeoniflorin, has a good effect on improving RA animal models. However, whether the anti-arthritis effect of CP-25 is related to Ahr remains unclear. METHODS CP-25 treatment ameliorated adjuvant-induced arthritis (AA), a mouse model of RA, by inhibiting Ahr-related activities in fibroblasts like synoviocytes (FLS). AA rats were treated with CP-25 or paroxetine from day 17 to 33 after immunization. RESULTS CP-25 alleviated arthritis symptoms and the pathological changes, decreased the expression of Ahr in the synovium and FLS of AA rats. Besides, treatment with CP-25 reduced the proliferation and migration of MH7A caused by Ahr activation. In addition, we also demonstrated that CP-25 down-regulated the co-expres?sion and co-localization of Ahr and G protein-coupled receptor kinase 2 (GRK2) in MH7A. CONCLUSION The data pre?sented here demonstrated that CP-25 suppressed FLS dysfunction in rats with AA, which were associated with reduced Ahr activation and the interaction between Ahr and GRK2.

Targeted inhibition of GRK2 kinase domain by CP-25 to reverse fibroblast-like synoviocytes dysfunction and improve collagen-induced arthritis in rats

Rheumatoid arthritis (RA) is an autoimmune disease and is mainly characterized by abnormal proliferation of fibroblast-like synoviocytes (FLS). The up-regulated cellular membrane expression of G protein coupled receptor kinase 2 (GRK2) of FLS plays a critical role in RA progression, the increase of GRK2 translocation activity promotes dysfunctional prostaglandin E4 receptor (EP4) signaling and FLS abnormal proliferation. Recently, although our group found that paeoniflorin-6'-

Inmunopatología de la brucelosis osteoarticular / Immunopathology of osteoarticular brucelosis

La brucelosis es una de las enfermedades zoonóticas más importantes a nivel mundial capaz de producir enfermedad crónica en los seres humanos. La localización osteoarticular es la presentación más común de la enfermedad activa en el hombre. Sin embargo, algunos de los mecanismos moleculares implicados en la enfermedad osteoarticular han comenzado a dilucidarse recientemente. Brucella abortus induce daño óseo a través de diversos mecanismos en los cuales están implicados TNF-α y RANKL. En estos procesos participan células inflamatorias que incluyen monocitos/macrófagos, neutrófilos, linfocitos T del tipo Th17 y linfocitos B. Además, B. abortus puede afectar directamente las células osteoarticulares. La bacteria inhibe la deposición de la matriz ósea por los osteoblastos y modifica el fenotipo de estas células para producir metaloproteinasas de matriz (MMPs) y la secreción de citoquinas que contribuyen a la degradación del hueso. Por otro lado, la infección por B. abortus induce un aumento en la osteoclastogénesis, lo que aumenta la resorción de la matriz ósea orgánica y mineral y contribuye al daño óseo. Dado que la patología inducida por Brucella afecta el tejido articular, se estudió el efecto de la infección sobre los sinoviocitos. Estos estudios revelaron que, además de inducir la activación de estas células para secretar quemoquinas, citoquinas proinflamatorias y MMPs, la infección inhibe la muerte por apoptosis de los sinoviocitos. Brucella es una bacteria intracelular que se replica en el retículo endoplásmico de los macrófagos. El análisis de los sinoviocitos infectados con B. abortus indicó que las bacterias también se multiplican en el retículo endoplasmático, lo que sugiere que la bacteria podría usar este tipo celular para la multiplicación intracelular durante la localización osteoarticular de la enfermedad. Los hallazgos presentados en esta revisión intentan responder a preguntas sobre los mediadores inflamatorios implicados en el daño osteoarticular causado por Brucella. (AU)

Brucellosis is one of the most important zoonotic diseases that can produce chronic disease in humans worldwide. Osteoarticular involvement is the most common presentation of human active disease. The molecular mechanisms implicated in bone damage have started to be elucidated. B. abortus induces bone damage through diverse mechanisms in which TNF-α and RANKL are implicated. These processes are driven by inflammatory cells, including monocytes/macrophages, neutrophils, Th17 lymphocytes and B cells. Also, Brucella abortus (B. abortus) can directly affect osteoarticular cells. The bacterium inhibits bone matrix deposition by osteoblast and modifies the phenotype of these cells to produce matrix methalloproteinases (MMPs) and cytokine secretion that contribute to bone matrix degradation. B. abortus also affects osteoclast increasing mineral and organic bone matrix resorption and contributing to bone damage. Since the pathology induced by Brucella species involves joint tissue, experiments conducted in sinoviocytes revealed that besides inducing the activation of these cells to secrete chemokines, proinflammatory cytokines and MMPS, the infection also inhibits sinoviocyte apoptosis. Brucella is an intracellular bacterium that replicate in the endoplasmic reticulum of macrophages. The analysis of B. abortus infected sinoviocytes indicated that bacteria also replicate in their reticulum suggesting that the bacterium could use this cell type for intracellular replication during the osteoarticular localization of the disease. The findings presented in this review try to answer key questions about the inflammatory mediators involved in osteoarticular damage caused by Brucella. (AU)

The study of the effect of Focal adhesion kinase and p53 in rheumatoid arthritis fibroblast like synoviocytes on cell proliferation / 中华风湿病学杂志

Objective To investigate the expression of Focal adhesion kinase (FAK) and p53 in rheumatoid arthritis (RA) Fibroblast Like Synoviocytes and to explore the pathogenesis of RA.This study also studied the effect of FAK and p53 on the proliferation of Fibroblast Like Synoviocytes in RA on order to identify new target for the treatment of RA.Methods Synovial tissue from RA patients were cultured in vitro;FLS were cultured in TNF-α (10 ng/ml) and different density of protease inhibitor (MG-132) (1,5,10,20 μmol/L)for 48 h.Then the level of mRNA of both FAK and p53 in each group was tested by real time polymerase chain reaction (RT-PCR).FLS were cultured with different density of protease inhibitor (MG-132) (1,5,10,20μmol/L) for 24 h,48 h,72 h,96 h then the cell proliferation of each group were tested.ANOVA was used to compare the means between groups.Results ① After TNF-α stimulating,the level of FAK mRNA was higher than the control group [(1.48±0.09 vs(1.03±0.33),F=7.807,P＜0.05).Compared with the control group,the level of p53 mRNA decreased [(0.97:±0.03) vs (1.30±0.39),F=19.933,P＞0.05).② After stimulated with different density of MG-132,the level of p53 mRNA was higher than the control group [(3.12±0.72) vs (1.30±0.39),F=19.933,P＜0.05),and the 5μ mol/L group was the highest of them.Compared with the control group,the level of FAK mRNA decreased [(0.93±0.20) vs (1.03±0.33),F=7.807,P＞0.05).③ After stimulated with different density of MG-132,the proliferation of FLS was slower than the control group (24 h F=16.654,P＜0.05;48 hF=13.652,P＜0.05;72 h F=72.999,P＜0.05;96 h F=51.533,P＜0.05).Conclusion In vitro,after stimulated with TNF-α,the level of mRNA of Focal adhesion kinase is increased,while the level of that decreases after MG-132 stimulation.Focal adhesion kinase is involved in the pathogenesis of rheumatoid arthritis.In vitro,after MG-132 stimulating,the level of mRNA of p53 is increased.p53 inhibits the excessive proliferation of RA synovial fibroblast cells and plays an important role in the pathogenesis of rheumatoid arthritis.

Influencing factors on the primary cultur e of mouse fibroblast-like synov iocyt es / 医学研究生学报

Obej ctive Knockout mice are widely used in the studies of joint diseases .This article investigated the effects of joint processing methods , collagenase types ,and collagenase digestion time on the number of primary fibroblast -like synoviocytes (FLSs) obtained from mice. Methods The hind legs of 6 of the 12 male mice were cut open from the hip joints , but not those of the other 6.FLSs were isolated using the type-Ⅳcollagenase digestion method and purified by differential digestion .Cell morphology was observed under the inverted microscope .The type, viability, and purity of the cells were determined by flow cytometry . Rse ults Significantly fewer FLSs were obtained from the mice with the hind legs cut open ( 19 133 ±115 ) than from those without (24 933 ± 503) (P<0.05).The numbers of FLSs collected from the cell suspension at 1, 2, 3, 4, 5,6 , and 7 hours after digestion were 700 ±300 , 600 ±100 , 15 200 ±900 , 5100 ±800 ,2700 ±300 , 900 ±200, and 300 ±100, respectively, the highest at 3 hours. There were statistically significant differences in the total number of FLSs obtained by type-Ⅳ and type-Ⅱ collagenase digestions (24900 ±500v s 18 100 ±400, P<0.05). Conclusion For in virt o culture of primary mouse FLSs, it is recommended that the hip joints be not cut open, and type-Ⅳcollagenase be used with cell sus-pension at 2-6hours after digestion .

Application of Induced Pluripotent Stem Cells in Rheumatology

Since induced pluripotent stem cell (iPSC) was first introduced by Yamanaka in 2006, it took only six years to win a Nobel Prize for his pioneering work. It is unusual to win a Nobel Prize for such recent research with a short history. Many scientists and clinicians are interested in iPSC for its potential application. Significant progression in this field has been made, while there remain many hurdles to overcome for application of iPSC technique in real clinics. In this review, the concept of reprogramming and the basic techniques of iPSC generation will be discussed for the reader's convenience, followed by discussion of recent progress, followed by the topics of "disease modeling" and "cell therapy" with iPSC in the second half of this article. Several examples of rheumatologic application of iPSC will be provided in the main text. If rheumatologists could understand the merits and potentials of iPSC, opportunities for innovative research and therapy can be expanded.

Mechanism of TWEAK on the synthesis of MMP-9 in fibroblast-like synoviocytes of rheumatoid arthritis / 中华微生物学和免疫学杂志

Objective To discuss the role of p38MAPK signal pathway in the process of TWEAK inducing rheumatoid arthritis(RA) fibroblast-like synoviocyte(FLS) to synthesize MMP-9 and look for a new target for RA treatment. Methods RA FLS were primarily cultured and stimulated with TWEAK. Western blot was used to detect the expression of p-p38MAPK and p65 in RA FLS. FLS were pretreated by SB203580 or not. ELISA was used to detect the concentration of MMP-9 in cell-cultured fluid. The mRNA expression of MMP-9 was measured by RT-PCR. Results TWEAK( 100 ng/ml) can make p38MAPK phosphorylated and increased the expression of p65 protein in the cell nucleus. SB203580 can partially inhibit the expression of MMP-9 and MMP-9 mRNA produced by RA FLS which is induced by TWEAK. Conclusion TWEAK induced RA FLS to synthesize MMP-9, in that process, the p38MAPK signal trausduction pathway was in active state, and induced the expression of NF-κB.

Study on the Apoptosis of Rheumatoid Arthritis Synoviocyte Induced by Arsenic Trioxide / 中国康复理论与实践

@# Objective To study the apoptosis of rheumatoid arthritis synoviocyte induced by arsenic trioxide.Methods Human rheumatoid arthritis fibroblast-like synoviocytes (HFLS-RA) were divided into control group and arsenic trioxide group. After 72 hours, pathology changes were observed and MTT tests were used to investigate apoptosis levels induced by arsenic trioxide.Results Arsenic trioxide can induce the apoptosis of rheumatoid arthritis synoviocyte, depending on time and dose.Conclusion Arsenic trioxide may induce apoptosis of rheumatoid arthritis synoviocyte.

beta ig-h3-Mediated Adhesion of Fibroblast-Like Synoviocytes in Rheumatoid Arthritis / 대한류마티스학회지

OBJECTIVE: beta ig-h3 is an extracellular matrix protein, which is overexpressed in synovial tissues of rheumatoid arthritis (RA) similar to adhesive glycoproteins. We sought to evaluate the compensatory role of beta ig-h3 with adhesive glycoproteins in mediating the adhesion of fibroblast- like synoviocytes (FLS) and to confirm the inhibitory effect of YH18 peptide of the 2nd fas-1 domain in beta ig-h3-mediated adhesion. METHODS: The adhesion of FLS isolated from synovial tissues of RA, was evaluated in 96 well microtiter plate coated with matrix proteins. Inhibitory effect of YH18 peptides from the 2nd and 4th fas-1 domains was estimated in beta ig-h3-mediated adhesion of FLS. RESULTS: The adhesion of FLS on beta ig-h3 was weaker than that of fibronectin and vitronectin. The beta ig-h3-mediated adhesion was enhanced by the stimulation with phorbol myristate acetate (PMA), but not by cytokines and growth factors. Combination of fibronectin with beta ig-h3 synergistically enhanced the adhesion of FLS, in contrast to the additive effect of vitronectin combined with beta ig-h3. YH18 peptide of the 2nd fas-1 domain did not block the beta ig-h3-mediated adhesion of FLS. CONCLUSION: Our results reveal that beta ig-h3 may regulate the adhesion of FLS through the interaction with adhesive glycoproteins and confirm that the essential motifs mediating adhesion on beta ig-h3 are different according to the type of cells.

Expression and Function of Calcineurin in Inflammatory Arthritis

BACKGROUND: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. METHODS: Immunohistochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1beta and TNF-alpha in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. RESULTS: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proinflammatory cytokines such as IL-1beta and TNF-alpha. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. CONCLUSION: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

Regulation of betaig-h3 Production in Rheumatoid Synovitis by Inflammatory Mediators / 대한류마티스학회지

OBJECTIVE: To investigate the expression pattern of transforming growth factor-beta-inducible gene-h3 (betaig-h3) within rheumatoid synovial tissue and the regulation of betaig-h3 synthesis in fibroblast-like synoviocyte (FLS). METHODS: Synovial tissues obtained from patients with rheumatoid arthritis and osteoarthritis were obtained during joint replacement surgery. betaig-h3 expression was evaluated with immunohistochemical stain. FLS was isolated from synovial tissues and stimulated with cytokines including TGF-beta, TNF-alpha, IL-1beta, IFN-gamma, IL-6, IL-4, and IL-10. betaig-h3 synthesis was measured using semiquantitative RT-PCR, ELISA, immunofluorescence stain, and flow cytometry. RESULTS: Expression of betaig-h3 was diffuse and abundant in both lining and sublining layers of rheumatoid synovium, which was more prominent than those of osteoarthritis. Production of betaig-h3 in FLS was regulated by TGF-beta1 in a dose-dependent manner and was highest at 5 ng/mL of TGF-beta1. TNF-alpha and IL-1beta upregulated the production of betaig-h3 from FLS synergistically with TGF-beta1 but other cytokines such as IL-4, IL-6, IL-10 did not affect. betaig-h3 synthesis was efficiently inhibited by dexamethasone at higher dose (100 nM) but not by cyclosporine-A. CONCLUSION: Production of betaig-h3, which is highly upregulated in rheumatoid synovitis, is differentially regulated by inflammatory cytokines.

Reciprocal Activation of Fibroblast-like Synoviocyte and Type II Collagen-reactive T cell in Rheumatoid Arthritis / 대한류마티스학회지

OBJECTIVE: To investigate the interaction between type II collagen (CII)-reactive T cell and fibroblast-like synoviocyte in rheumatoid arthritis (RA). METHODS: Peripheral blood T cells from RA patients were cultured with bovine CII and analyzed by flow cytometry. After co-culture with CII-reactive T cells and fibroblast-like synoviocytes (FLS), the expression of cytokines (IL-15 and TNF-alpha from FLS, IFN-gamma and IL-17 from CII-reactive T cells) were determined by ELISA and RT-PCR. RESULTS: CII-reactive T cells expressed CD69, one of the early activation markers, and produced significant amount of IFN-gamma, and proliferated. IL-15 and TNF-alpha expression from FLS were significantly elevated when co-culture with CII-reactive T cells and inhibited by physical interruption of cell-to-cell contact or anti-CD40 antibody. IFN-gamma and IL-17 expression from CII-reactive T cells were also significantly elevated when co-culture with FLS and inhibited by anti-IL-15 monoclonal antibody. CONCLUSIONS: CII-reactive T cells can activate FLS to secret proinflammatoy cytokines and interactions between these two cells drive further activation of each other. These data suggest that CII-reactive T cell may play a important role in pathogenesis of RA.

Highly Efficient Gene Expression in Rabbit Synoviocytes Using EBV-Based Plasmid

BACKGROUND: Rheumatoid arthritis (RA) is an autoimmune disorder characterized by chronic synovial inflammation which leads to joint destruction. Gene therapy of RA targets the players of inflammation or articular destruction. However, viral vectors have safety problems and side effects, while non-viral vectors suffer from inefficient gene transfer and fast loss of gene expression. To overcome the limits of non-vial vectors, an EBV-based plasmid which is known to exert prolonged high level gene expression can be used. METHODS: pEBVGFP, pEBVIL-10, and pEBVvIL-10 were constructed by cloning GFP, IL-10, and vIL-10 genes into an EBV-based plasmid, respectively. The pGFP was used as a control plasmid. Each constructs were lipofected into HIG-82 rabbit synoviocytes. The expression of GFP was monitored by FACS and confocal microscopy. IL-10 and vIL-10 expressions were measured by ELISA. RESULTS: GFP expression 2 days after transfection was achieved in 33.2% of cells. GFP-expressing cells transfected with pGFP decreased rapidly from 4 days after transfection and disappeared completely by 11 days. Cells transfected with pEBVGFP began to decrease slowly from 4 days. But GFP expression was detected for over 35 days. In addition, HIG-82 cells transfected with pEBVIL-10 (44.6+/-1.5 ng/ml) or pEBVvIL-10 (51.0+/-5.7 ng/ml) secreted these cytokines at high levels. High level cytokine production by hygromycin selection was maintained at least for up to 26 days after transfection. CONCLUSION: These results suggest that the EBV-based plasmid has a potential to improve non-viral gene transfer system and may be applicable to treat RA without the drawbacks of viral vectors.

An Immuno -Electron Microscopic Study for Perlecan in the Synovium of Degenerative Arthritis of Knee / 대한해부학회지

This study was designed to observe the expression of perlecan in the normal and degenerative arthritic synovial membrane. By using the immunohistochemical staining and immuno -electron microscopical gold labeling techniques, we observed five materials of normal and degenerative arthritic synovia each. The results were as follows. 1. By the immunohistochemical methods, perlecan -positive staining was seen on the 1 ~2 cell layers of the normal synovial membrane. But, a weaker staining compared to that seen in the normal synovial membrane was found in the degenerative arthritic synovial membrane. 2. Under the electron microscopic observation, perlecan was largely distributed in the rough endoplasmic reticulum of the secretory synovial cell, and in the vacuoles of the phagocytic synovial cell on the normal synovium of the human knee joint. It was also found in the extracellular matrix of the synovial membrane. 3. Perlecan -positive cells were also identified on the degenerative arthritic synovium of the human knee joint. However, fewer perlecan was observed here than that found in the normal synovium. In conclusion, perlecan is synthesized by the secretory synovial cells and degraded by the phagocytic synovial cells. And it, known as a major component of the basement membrane, also proven to exist in the extracellular matrix of the synovial membrane having no basement membrane. From the fact that less perlecan was observed in the degenerative arthritis, perlecan is might to play a major role in the degenerative process.

EFFECT OF TOTAL FLAVONOIDS OF CHRYSANTHEMUM INDICUM ON THE APOPTOSIS OF ADJUVANT ARTHRITIS RAT SYNOVIOCYTES AND THE EXPRESSION OF CASPASE-3 / 解剖学报

Objective To study the effect of the extract of total flavonoids of Chrysanthemum indicum(TFC) on adjuvant arthritis(AA) synoviocytes.Methods Totally 0.1ml of the complete Freund's adjuvant was subcutaneously injected into the right hind feet pads of 20 SD rats.24 days after immunity synoviocytes in the knee joint were treated with TFC.Cell morphology was examined with electron microscopy.Protein level of caspase-3 cleaved fragments was analyzed by Western blotting.The annexin V stain assay was applied to explore the effect of caspase-3 inhibitor on the amelioration in synovial cells apoptosis of AA rats.Results Typical morphology and biochemical feature of apoptosis in synovial cells of AA rats were observed with TFC.The protein level of caspase-3 cleaved fragments increased obviously and was related with the concentration of TFC in synovial cells of AA rats.The apoptotic cells positively stained with annexin V were markedly reduced by caspase-3 inhibitor.Conclusion TFC can induce apoptosis in AA rats synoviocytes,which may achieve therapeutical effects in AA.The activation of caspase-3 may be one of the main causations.

In Vitro Effects of Ultrasound on Fibroblast Like Synoviocytes from Rheumatoid Arthritis

OBJECTIVE: Ultrasound has been therapeutically applied for pain control in rheumatoid arthritis although little physiologic effects of sonication on rheumatoid tissue were known. This investigation was conducted to determine the effects of sonication on the cell proliferation and matrix metalloproteinase (MMP) production of cultured fibroblast like synoviocytes (FLS) derived from synovial tissues of rheumatoid arthritis. METHOD: Pulsed ultrasound (1.0 MHZ, 20 msec on, 80 msec off) with varying intensities (0, 0.1, 0.25, 0.5, 0.75, 1.0 W/cm2) was applied to experimental cell groups growing as monolayers in culture plates for varying durations (0, 30, 90, 180 seconds) in the presence and absence of interleukin-1beta (IL-1beta). RESULTS: There were no significant differences in thymidine incorporation between 0, 30, 90 and 180 second sonication groups with 0.5 W/cm2 after 1 day and 2 days. There were no significant differences in thymidine incorporation between 0, 0.1, 0.25, 0.5, 0.75, 1.0 W/cm2 sonication groups 1 day and 2 days after 90 second sonication. There were significant increase in MMP-1 (p=0.025) and MMP-3 production (p=0.000) of FLS after sonication in the absence of IL-1beta but there were no significant differences in MMP-1 and MMP-3 production in the presence of IL-1beta. And MMP-1 and MMP-3 production were increased significantly in the presence of IL-1beta but not than in the absence of IL-1beta. CONCLUSION: While comparisons made between a limited number of FLS cell lines must be open to question, the overall consistency of the findings suggest sonication with nonthermal effect is not the contraindication in rheumatoid arthritis treatment but further study is needed in vivo in animal and in clinical studies.

THE ROLE MONOCYTE/MACROPHAGE IN THE PATHOGENESIS OF DIALYSIS RELATED AMYLOIDO-SIS / 解放军医学杂志

The pathogenesis of dialysis related amyloidosis, which occurs preferentially in osteo articular tissues, is still incom pletely understood. Although recent histological studies have shown the accumulation of monocytes/macrophages around amyloid deposits, the factor(s) causing their infiltration and pathological involvement have yet to be fully elucidated. The present studies demonstrate that ? 2 microglobulin (? 2 m), the major constituent protein in amyloid fibrils, can be modified in situ by advanced glycation end products (AGE) through binding to AGE modified collagen. AGE ? 2 m attracts monocytes via direct chemotaxis and through regulation of synoviocyte derived chemokine. AGE modified ? 2 m significantly delays spontaneous apoptosis of human monocytes via a pathway mediated by the receptor for AGE (RAGE), processes which may increase the accumulation of inflammatory monocytes. In addition to recruit monocytes, AGE ? 2 m stimulates macrophages to release IL 1?, TNF ? and IL 6.These proinflammatory cytokines upregulate the expression of adhesion molecules such as ICAM 1 and VCAM 1 by synovial cells and induce the release of synoviocyte derived collagenase which may contribute to the degradation of matrix. These AGE ? 2 m induced perturbation of monocytes and cellular inflammatory reactions eventually result in osteo articular tissue damage and destruction seen in DRA.

Antioxidative effect of total axtract of astragalosides / 中国临床药理学与治疗学

Aim The relationship between the therapeutic effect of TEA (total extract of astragalosides) on adjuvant arthritic (AA) rats and its antioxidative effects were studied . Methods The volume of non-injected hind paw of AA rats and malondialdehyde(MAD) content of arthritic synoviocytes from AA rats were measured and the proliferative responses of fibroblast, the level of superoxide anion() and the hydroxyl radical (?OH) generated in vitro were detected . Results The anti-inflammatory effect of TEA might be related to its antioxidative activity.In vitro low-level oxidative stress promoted the proliferative responses of fibroblast in rats synovium, which was marked by inhibited by TEA in a concentration-dependant manner. Further study showed that TEA could inhibit NBT reduction induced by both xanthine-xanthine oxidase and non-enzyme generated , but the inhibitory effect of this compound on activity of xanthine oxidase was obtained only at high concentration (more than 80 ?g?ml-1). It was also found that TEA could dose-dependantly inhibit the hydroxylation of benzoic acid induced by Fenton reaction generated ?OH. Conclusion TEA has significant therapeutic effects on AA rats, which might be related to its antioxidative effects

Nitric Oxide(NO) in Inflammatory Arthritis / 대한내과학회지

OBJECTIVES:Nitric Oxide(NO) is a toxic, inorganic, gaseous free radical produced during the metabolism of L-Arginine by NO synthase(NOS). It has been implicated in a rapidly growing number of physiological and pathophysiological processes such as cytotoxic effects against microbes and tumor cells, blood vessel dilation and neurotransmitter. Recently there is growing evidence implicating NO in immune regulation, inflammation, autoimmunity, and arthritis. We performed this study to determine a role for nitric oxide in inflammatory arthritis especially rheumatoid arthritis(RA). METHODS: We measured (1) the concentrations of nitrite, a breakdown product of nitric oxide, in serum and synovial fluid from patients with RA and osteoarthritis(OA) and in the serum of controls (2) the concentrations of nitrite in the supernatant of cultured synovial tissue with RA and OA and (3) determined whether human chondrocytes and synoviocytes can synthesize nitric oxide and if so, how production is regulated by cytokines and antirheumatic drugs. RESULTS: 1) Serum nitrite concentrations in patients with RA and OA were higher than in controls. In both disease groups synovial fluid nitrite was higher than serum nitrite. Serum and synovial fluid nitrite concenrations in RA were higher than those in OA. However, those findings are not statistically significant. 2) Although these findings are not statistically significant, the concentration of nitrite in the supernatant of cultured synavial tissue with RA was higher than that in OA. 3) IL-1beta and TNF-alpah induced the biosynthesis of NO by chondrocytes and synoviocytes. IGF-1 and TGF-beta failed to provoke the production of NO. The biosynthesis of NO required an induction period of approximately 6 hours and was inhibited by L-NMMA and cycloheximide. Dexamethasone, indomethacin, gold sodium thiomalate and methotrexate had no effect on the induction of NO biosynthesis. CONCLUSION: These results suggest a role for nitric oxide as an inflommatory mediator in inflammatory arthritis.

The Roles of Intercellular Adhesion Molecule I in T Cell Adhesion Tosynovial Cell in Patients with Rheumatoid Arthritis / 대한류마티스학회지

OBJECTIVE: It has previously been shown that cellular interaction between infiltrating mononuclear cells and synoviocyte are important in the initiation and perpetuation of autoimmune processes in the synovial tissue of patients with rheuamtoid arthritis. Thus, we have investigated the molecular basis of T cell-synovial cell interaction in the cultured synoviocytes from patients with rheumatoid arthritis. METHODS: Using an immunohistochemical staining technique and inhibition study with monoclonal antibody, we studied the expression and the function of intercellular adhesion molecule 1 (ICAM-1) in T cell-synovial cell interaction in the cultured synoviocytes. RESULTS: Expression of ICAM-1 was diffusely observed in most components of rheumatoid synovium and readily up-regulated by IL-1. T cell-synovial cell interaction was inhibited by monoclehal antibody aganinst ICAM-1. CONCLUSION: These results showed that ICAM-1 was involed in the cellular interaction between T lymphocytes and synovial cells in patients with rheumatoid arthritis.