尿激酶型纤溶酶原激活物受体在类风湿关节炎滑膜组织细胞的表达

目的观察尿激酶型纤溶酶原激活物受体（uPAR）在类风湿关节炎（an）滑膜组织细胞膜（muPAR）和细胞质（cuPAR）的表达及分布。方法分别采用间接免疫荧光和免疫组化等方法检测18份RA、10份骨关节炎（OA）和4份正常滑膜组织中uPAR的表达及分布。结果RA滑膜组织细胞中muPAR阳性表达率为（66．0±9．4）％,主要分布于滑膜组织中血管翳周围,如血管内皮细胞和间质细胞等;而cuPAR阳性表达率为（61．0±5．8）％,主要分布于滑膜衬里层和滑膜下层细胞,如滑膜下层巨噬细胞样细胞、滑膜衬里层细胞及单个核白细胞等。OA滑膜组织中uPAR也呈不同程度的阳性表达,但与RA相比阳性表达程度较弱,阳性细胞数量较少。相关性分析：muPAR、cuPAR与RA滑膜炎积分呈正相关（r分别为0．672、0．649,P〈0．01）。结论muPAR表达阳性的细胞可能主要参与RA血管翳的形成及滑膜组织的侵袭等病理过程;而cuPAR表达阳性的细胞则可能与炎性介质的释放和致炎作用有关;提示uPAR在RA的病理过程中起重要作用。     

类风湿关节炎滑膜组织PDGF与TGF-β1的表达与意义

应用免疫组织华沙SABC对24列类风湿关节炎（RA）及4例正常滑膜组织中血小板源性生长因子（PDGF），转化生长因子β1（TGF－β1）的表达分布情况进行观察分析，结果发现20例RA滑膜组织衬里层的巨噬细胞样细胞，浸润的炎性细胞与血管内皮细胞，成纤维细胞可见PGDF的表达分析，且巨噬细胞样细胞，浸润的炎症细胞的阳性程度强于血管内皮细胞，成纤维细胞。22例RA滑膜组织衬里层与衬里下层的成纤维细胞，巨噬细胞样细胞，血管内皮细胞均见TGF－β1的阳性表达分布。两者比较，阳性细胞的染色程度，染色细胞数与分布范围大致相等。4例正常滑膜组织中PDGF，TGF－β1的表达分布均为阴性。认为RA滑膜组织中PDEFTGF－β1的表达分布较正常多且表达程度大致相等，两者协同作用，参与了RA病理过程的滑膜衬里层增生，炎性细胞浸润，血管增殖与滑膜血管翳的形成。     

CD147分子和基质金属蛋白酶在类风湿关节炎滑膜中的表达

目的 探讨类风湿关节炎(RA)滑膜组织CD147的表达及其与基质金属蛋白酶(MMP)-01和MMP-2表达的相关性。方法 采用免疫组织化学SP(streptavidin／peroxidase)染色方法检测11例RA患者受损关节软骨-血管翳接合部（CPJ)滑膜组织中CD147和MMP-1及MMP-2的表达，并与3例骨关节炎（OA)患者滑膜组织CD147和MMP-1及MMP-2的检测相对照。结果 3例OA滑膜组织CD147和MMP-1及MMP-2的表达均为阴性，而11例RA滑膜组织中均有CD147和MMP-1及MMP-2的表达。其中，表达CD147的细胞为单核-巨噬细胞、淋巴细胞和滑膜成纤维样细胞，表达MMP-1及MMP-2的细胞为滑膜成纤维样细胞。统计学分析表明RA滑膜细胞CD147的表达和MMP-1及MMP-2的表达间存在显著相关性。结论 CD147在RA滑膜组织中表达增高，可能是导敏RA受损关节软骨、骨基质降解的重要因素之一。     

类风湿关节炎中尿激酶型纤溶酶原激活物及其受体蛋白和基因的表达

目的　检测尿激酶型纤溶酶原激活物 (uPA)及其受体 (uPAR)蛋白和mRNA在类风湿关节炎 (RA)的表达 ,探讨uPA、uPAR基因在RA细胞外基质降解中的作用。方法　采用免疫组化和cDNA mRNA原位分子杂交技术分别检测了 2 4例RA、18例骨关节炎 (OA)和 6例正常滑膜组织中uPA、uPAR蛋白和mRNA的分布及表达情况。结果　 2 4例RA滑膜组织均呈uPA、uPAR蛋白和mRNA的阳性表达 ,uPA、uPAR蛋白的强阳性率高于mRNA。uPA、uPAR蛋白和mRNA阳性信号主要分布在RA滑膜衬里细胞、滑膜下层单核细胞、巨噬细胞样细胞及血管内皮细胞 ;18例OA滑膜组织中 ,uPA、uPAR蛋白和mRNA的表达部位类似于RA ,但阳性率、阳性程度及分布范围均明显低于RA滑膜组织 ,两组之间蛋白和mRNA表达的差异均有显著性 (P <0 0 1或P <0 0 0 1)。 6例正常滑膜组织呈阴性反应。结论　RA滑膜组织存在高水平uPA、uPAR蛋白和mRNA的表达 ,提示在RA的发生发展过程中 ,uPA和uPAR基因起着重要作用 ;RA和OA中uPA、uPAR基因表达水平的差异 ,可能与这两种疾病软骨和骨基质降解的程度及进程等临床表现密切相关     

老年骨关节炎患者关节软骨和滑膜中转化生长因子β及受体和金属蛋白酶组织抑制物的研究

目的 探讨骨关节炎（OA）患者关节软骨和滑膜中转化生长因子β（TGFβ1）、转化生长因子β受体（TGFβR）和金属蛋白酶组织抑制物－1（TIMP－1）变化及其与OA发病的关系。方法 采用免疫组化方法检测23例老年OA患者及3例外伤患者正常关节软骨和滑膜TGFβ1、转化生长因子βI类受体（TGFβRI）、转化生长因子βⅡ类受体（TGFβRⅡ）和TIMP－1的分布和阳性程度。结果 OA患者中14例关节软骨和16例滑膜TGFβ1染色呈阳性或弱阳性,14例关节软骨和15例滑膜TIMP－1染色呈弱阳性,全部OA患者关节软骨和滑膜TGFβRI和TGFβRⅡ染色呈强阳性。阳性细胞包括软骨细胞、滑膜衬里细胞、滑膜下层的血管内皮细胞和间质巨噬细胞等。结论 老年OA患者关节软骨和滑膜中TGFβ1、TGFβ及TIMP－1的变化可能与OA发病有一定的关系。     

护骨素在强直性脊柱炎外周关节骨质破坏病理机制中的作用

目的检测护骨素(OPG)在强直性脊柱炎(AS)外周关节滑膜组织中的表达,并以类风湿关节炎(RA)、骨关节炎(OA)患者和健康志愿者外周关节滑膜组织为对照,了解OPG表达与AS患者外周关节骨质破坏病理改变的相关性。方法应用单克隆抗体,通过免疫组织化学方法检测13例AS、16例RA、17例OA及6名健康对照关节滑膜组织中OPG的表达及分布状况,并通过计算机辅助图像分析系统和半定量分析方法确定OPG在各滑膜组织中表达水平之间的差异,分析OPG表达与炎性指标及关节X线分期之间的相关性。结果OPG蛋白在所有13例AS滑膜组织中均有阳性表达,阳性细胞主要分布于滑膜衬里层、衬里下层区域,滑膜软骨交界区OPG表达明显低于滑膜衬里层和衬里下层;2名健康对照滑膜组织中有阳性表达,但明显低于AS患者组。RA及OA患者组未见OPG阳性表达。结论譹AS组滑膜组织中OPG表达水平明显高于健康对照组(P<0.01),而RA、OA滑膜组织中未见OPG表达,说明OPG的高水平表达是滑膜组织对炎症反应/关节破坏所特有的表现,OPG的局部表达是维持AS关节骨代谢稳定的重要因素,这可能是大多数AS患者外周关节受累预后好于RA的原因之一。譺AS患者组滑膜软骨交界区中OPG表达量显著减少可能是导致关节骨质破坏的重要原因,提示关节局部应用OPG治疗有可能改善受累关节的骨     

趋化因子CXCL16/CXCR6及其在类风湿关节炎发病中的作用及意义

类风湿关节炎(rheumatoid arthritis,RA)是一种主要累及滑膜关节的自身免疫性疾病,其病理改变包括慢性滑膜炎、血管翳形成及大量炎细胞浸润等.滑膜中大量聚集的炎细胞可产生和释放大量炎性介质,破坏关节软骨和骨,最终导致关节破坏和畸形.趋化因子在炎细胞向滑膜组织迁移及活化过程中发挥了关键作用,尤其是白发现CXCL16在RA滑膜中高表达以来,其在RA发病机制中的作用越来越受到重视,且可能成为RA治疗的新靶点.以下就CXCL16/CXCR6及其在RA发病中的作用和意义进行综述.     

骨桥蛋白在类风湿关节炎发病机制中的研究进展

类风湿关节炎(rheumatoid arthritis,RA)为慢性炎症性自身免疫病,主要病理特点为滑膜炎、血管翳的形成,以及软骨和骨的破坏.RA病变核心部位在关节滑膜,滑膜主要的组织细胞学改变有:滑膜内膜层细胞[成纤维样滑膜细胞(FLS)和巨噬样细胞]的增生、滑膜内膜下层炎性细胞(巨噬细胞、淋巴细胞等)的浸润,侵蚀关节面的血管翳形成,破骨细胞活化并介导矿化软骨和软骨下骨的侵蚀.病变中涉及到的分子有细胞因子、黏附分子、趋化因子、蛋白酶等.     

血管内皮细胞生长因子与类风湿关节炎发病机制及其治疗的相关性

血管内皮细胞生长因子(VEGF)及其受体以调控类风湿关节炎血管形成为最主要的特点。他对滑膜炎症的发展及滑膜血管翳的形成具有十分重要的作用。其在类风湿关节炎患者滑液和血清中的表达,与疾病严重程度呈相关性。因此,以VEGF及其受体为靶点,将成为治疗类风湿关节炎的新途径。     

类风湿关节炎滑膜组织中细胞外基质金属蛋白酶诱导因子的研究

目的研究类风湿关节炎(RA)患者滑膜组织中细胞外基质金属蛋白酶诱导因子(EMM-PRIN)的表达,探讨其在RA致病中的作用。方法对20例RA患者膝关节滑膜组织标本采用免疫组织化学染色和半定量反转录-聚合酶联反应(RT-PCR)法,观察RA滑膜组织中EMMPRIN的蛋白及mRNA表达。10例骨关节炎OA滑膜作为对照。结果RA滑膜内衬层的单核细胞、成纤维细胞中EMMPRIN阳性表达广泛,RA组EMMPRIN的免疫反应明显强于对照组(P<0.01)。14例RA、2例OA滑膜标本EMM-PRINmRNA表达阳性,EMMPRINmRNA表达水平RA也明显高于OA组。结论滑膜中过度表达的EMMPRIN通过上调MMPs促进关节软骨的破坏。因此选择性地抑制EMMPRIN生物活性,可能是治疗RA的新途径。     

EBV-encoded small RNA1 and nonresolving inflammation in rheumatoid arthritis

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by perpetuated inflammation in multiple joints. To date, there is no cure for RA, and the causal factor for non-resolving inflammation in RA remains unclear. In this study, we initially observed expression of Epstein–Barr virus-encoded small RNA1 (EBER1) in the synovial tissue of all five patients who showed nonresolving RA inflammation. By contrast, EBER1 was detected in the synovial tissue of only one out of seven patients with advanced osteoarthritis (OA; p < 0.01, Fisher’s exact test). To confirm this finding, we conducted a second study on synovial tissue samples taken from 23 patients with nonresolving RA inflammation and 13 patients with OA. All synovial samples from patients with nonresolving inflammation of RA showed positive expression of EBER1 (23/23, 100%), whereas none of the synovial samples from patients with OA showed expression of EBER1 (0/13, 0%; p < 0.001, by Fisher’s exact test). In vitro, transfection of RA synovial fibroblasts with EBER1 induced the production of interleukin-6. Taken together, these data strongly suggest that nonresolving RA inflammation is strongly related to the presence of EBER1, which might be, at least partially, responsible for synovial fibroblast interleukin-6 production.     

Expression of interferon beta in synovial tissue from patients with rheumatoid arthritis: comparison with patients with osteoarthritis and reactive arthritis

BACKGROUND: IFNbeta may have immunomodulatory effects in rheumatoid arthritis (RA) and its increased production in RA synovium may be a reactive attempt to inhibit inflammation. OBJECTIVE: To determine the expression of IFNbeta in the synovial tissue of patients with RA, osteoarthritis, and reactive arthritis. METHODS: Synovial biopsy specimens were obtained by arthroscopy from patients with RA and disease controls for immunohistological analysis using a monoclonal antibody specific for IFNbeta. Bound antibody was detected by an immunoperoxidase method. Stained sections were evaluated by computer assisted image analysis. Double stainings were performed with antibodies to detect CD55 positive fibroblast-like synoviocytes (FLS), CD68 positive macrophages, and CD83 positive dendritic cells (DCs) co-expressing IFNbeta. RESULTS: IFNbeta protein was abundantly expressed in the synovium of patients with RA. Digital image analysis showed a significant increase in the mean integrated optical density for IFNbeta expression in RA synovial tissue compared with disease controls. Specific up regulation of IFNbeta expression was also seen when the results were controlled for cell numbers. Phenotypic analysis showed that FLS, especially, but also macrophages and DCs may express IFNbeta in RA synovial tissue. CONCLUSIONS: The increased expression of IFNbeta in RA synovium suggests activation of an immunomodulatory mechanism that could inhibit synovial inflammation.     

Detection of major histocompatibility complex/human cartilage gp-39 complexes in rheumatoid arthritis synovitis as a specific and independent histologic marker

OBJECTIVE: Peptide 263-275 is the immunodominant epitope of human cartilage (HC) gp-39, a candidate autoantigen in rheumatoid arthritis (RA). We recently generated and characterized a monoclonal antibody (mAb) called 12A, which is directed against HLA-DR4/HC gp-39(263-275) complexes and inhibits specific T cell responses in vitro. The aim of the present study was to analyze whether presentation of the immunodominant epitope of HC gp-39 by shared epitope-positive synovial dendritic cells is a specific event in the development of chronic synovial inflammation in RA. METHODS: Staining with mAb 12A was performed on synovium obtained from clinically swollen joints in 65 patients with RA and 67 non-RA controls and from joints without clinical effusion in 9 additional patients with RA. RESULTS: Monoclonal antibody 12A staining was observed in the synovium of 40 of the 65 patients with RA. Histologically, expression of HC gp-39, lymphoid aggregates, CD3, and CD1a as well as the global inflammation score were higher in mAb 12A-positive RA synovium than in mAb 12A-negative synovium, indicating a follicular synovitis in these samples. Accordingly, mAb 12A stained dendritic cells in the close vicinity of lymphoid aggregates. No mAb 12A staining was detected in synovium obtained from RA joints without effusion. Clinically, there were no correlations between mAb 12A staining and clinical or biologic parameters in RA. However, positive staining was observed in 61.5% of the inflamed RA synovial samples compared with only 3.0% of the control samples (P < 0.001). This mAb 12A staining was not related to intracellular citrullinated peptides, which are another specific histologic marker for RA. CONCLUSION: Presentation by synovial dendritic cells of the immunodominant epitope of HC gp-39, in the context of the shared epitope, is associated with characteristic histologic features of follicular synovitis and is highly specific for RA. This suggests a contribution to the autoimmune-related tissue inflammation and provides a new and independent tool for the immunopathologic diagnosis of RA.     

Expression of the apoptosis accelerator Bax in rheumatoid arthritis synovium

OBJECTIVE: The aim of this study was to analyze the expression of apoptosis-related molecules in rheumatoid arthritis (RA) synovium, with special emphasis on the apoptosis accelerator Bax. METHODS: Immunohistochemical analysis of Bax, Bcl-2, and Bcl-x(L) was performed in tissue specimens of patients with RA and compared to normal synovial tissue. Expression of Bax was additionally determined by double labeling with CD68, p53, and Ki-67 (clone MIB-1). Apoptotic cells were further identified by the terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) method. RESULTS: In RA, expression of Bax was higher than in healthy controls and occurred in CD68-positive and -negative synoviocytes. Strong Bax staining was also found in chondrocytes at sites of cartilage degradation. Bax-positive synoviocytes could be detected with p53 and also with Ki-67. Bax and Bcl-x(L) were markedly colocalized in synovium. The TUNEL method revealed only few positive synoviocytes. CONCLUSIONS: The marked colocalization of Bax and antiapoptotic Bcl-x(L) as well as the low frequency of TUNEL-positive cells in RA synovium suggest that Bax activity is not sufficient to decrease synovial hyperplasia in RA. Apoptotic mechanisms in RA chondrocytes might also be important for the pathogenesis of joint damage.     

Expression of osteopontin messenger RNA and protein in rheumatoid arthritis: effects of osteopontin on the release of collagenase 1 from articular chondrocytes and synovial fibroblasts

OBJECTIVE: Osteopontin (OPN) is an extracellular matrix protein that has been implicated in the interactions between tumor cells and host matrix, including those involved in invasion and spread of tumor cells. Because joint destruction in rheumatoid arthritis (RA) is mediated by the invasive growth of synovial tissue through its attachment to cartilage, we examined the expression of OPN in the synovia of patients with RA and the effect of OPN on the production of collagenase 1 in rheumatoid synovial fibroblasts and articular chondrocytes. METHODS: The expression of OPN messenger RNA (mRNA) and protein in synovia from 10 RA patients was examined by in situ hybridization and immunohistochemistry. Synovial fibroblasts from RA patients and articular chondrocytes from patients without joint disease were cultured in the presence of various concentrations of OPN, and levels of collagenase 1 in the culture supernatants were measured by enzyme-linked immunosorbent assay. RESULTS: The expression of OPN mRNA and protein was observed in 9 of 10 specimens obtained from patients with RA. OPN was expressed in the synovial lining and sublining layer and at the interface of cartilage and invading synovium. Double labeling revealed that the majority of OPN-expressing cells were positive for the fibroblast-specific enzyme prolyl 4-hydroxylase and negative for the macrophage marker CD68, while only a few, single OPN-expressing cells were positive for CD68 at sites of synovial invasion into cartilage. OPN staining was not observed in lymphocytic infiltrates or leukocyte common antigen (CD45)-positive cells. Three of 3 cultures of human articular chondrocytes secreted detectable basal amounts of collagenase, with a dose-dependent increase upon OPN stimulation, while synovial fibroblast cultures produced much lower levels of collagenase, with only 2 of 4 fibroblast cultures responding in a dose-dependent manner. CONCLUSION: These findings suggest that OPN produced by synovial fibroblasts in the synovial lining layer and at sites of cartilage invasion not only mediates attachment of these cells to cartilage, but also contributes to matrix degradation in RA by stimulating the secretion of collagenase 1 in articular chondrocytes.