Serum MMP-3 and MMP-1 and progression of joint damage in early rheumatoid arthritis

OBJECTIVE: Expression and activation of matrix metalloproteinases such as MMP-3 (stromelysin-1) and MMP-1 (collagenase-1) are increased in patients with rheumatoid arthritis (RA). Previous negative reports of their value as predictors of joint damage may be due to the lack of a large longitudinal study of early RA patients. This study evaluated their use in assessing early untreated patients with RA and predicting subsequent joint damage. METHODS: Ninety-eight patients with early untreated RA of less than 12 months duration and 20 normal controls had baseline serum samples tested with a double-antibody enzyme-linked immunosorbent assay for each of MMP-1 and MMP-3. The subsequent changes in Larsen score (DeltaLarsen) and Health Assessment Questionnaire (DeltaHAQ) over the first 12 months were recorded. RESULTS: Baseline serum levels of MMP-3 and MMP-1 correlated significantly with baseline C-reactive protein (CRP) (r=0.42 and 0.49, P<0.001), DeltaHAQ (r=0.32 and 0.30, P<0.01) and DeltaLarsen (r=0.23 and 0.32, P<0.05) respectively. Analysis of the group of patients with a normal CRP at presentation (n=21) showed correlation of the baseline MMP-3 and MMP-1 with the presence of erosive disease during the first 12 months (r=0.52 and 0.65 respectively, P<0.05). Logistic regression analysis, in the patients who were non-erosive at presentation, showed that the strongest correlation with progression in Larsen score was the baseline MMP-3 level (r=0.30, P=0.01). CONCLUSIONS: Baseline serum MMP-1 and MMP-3 levels correlate with disease activity and predict functional and radiographic outcome in early untreated RA. They may have a particular value in predicting the progression of erosive disease in patients who are not erosive at presentation.     

Cordycepin inhibits IL-1{beta}-induced MMP-1 and MMP-3 expression in rheumatoid arthritis synovial fibroblasts

Objective. MMP is a key enzyme in the degradation of extracellularmatrices, and its expression plays important roles in inflammatorydiseases. Cordycepin (3'-deoxyadenosine), a bioactive compoundof Cordyceps militaris, has been shown to exhibit many pharmacologicalactivities, such as anti-cancer, anti-inflammatory and anti-infectionactivities. In this study, we aimed at the inhibitory effectof cordycepin on IL-1β-induced MMP-1 and MMP-3 expressionas well as the molecular basis using RA synovial fibroblasts(RASFs). Methods. RASFs were isolated from synovial tissue obtained from12 patients with RA and cultured in monolayer. Expression ofMMP-1 and MMP-3 was evaluated using western blotting and real-timePCR. Chemokines were analysed by ELISA. The phosphorylationof mitogen-activated protein kinase was measured by westernblotting. Electrophoretic mobility shift assay was performedto evaluate binding activities of DNA to nuclear factor-B (NF-B)and activator protein-1 (AP-1). Results. Cordycepin inhibited IL-1β-induced MMP-1 and MMP-3expressions in RASFs in a dose-dependent manner. Among variouschemokines [such as monocyte chemoattractant protein-1 (MCP-1),GRO-, regulated upon activation, normal T-cell expressed andpresumably secreted (RANTES) and epithelial neutrophil activatingpeptide 78 (ENA-78)], cordycepin specifically blocked IL-1β-inducedENA-78 production in RASF. Moreover, cordycepin significantlyinhibited IL-1β-induced p38/JNK and AP-1 activation, butnot extracellular signal-regulated kinase (ERK) and NF-B activation. Conclusions. Cordycepin is a potent inhibitor of IL-1β-inducedchemokine production and MMP expression and strongly blocksthe p38/JNK/AP-1 signalling pathway in RASFs. KEY WORDS: Cordycepin, Interleukin-1β, Matrix metalloproteinase, p38 mitogen-activated protein kinase, Activator protein-1     

Enhanced production of MMP-1, MMP-3, MMP-13, and RANTES by interaction of chondrocytes with autologous T cells

It has been reported that T cells and chondrocytes interact through cell surface molecules such as MHC, CD4 or CD8 in osteoarthritis (OA) and T cells are activated. The objective of this study is to investigate the responses of chondrocyte–T cell interaction in terms of metalloprotease (MMP) and chemokine production. Articular cartilage and autologous blood were obtained from patients with OA and fracture who under went prosthetic surgery. Synovial fluid (SF) was collected from OA patients. Isolated chondrocytes were co-cultured with autologous T cells. SF cells were analyzed by immunostaining or Alcian blue staining. The production of MMP-1, MMP-3, MMP-13, and regulated on activation, normal T expressed and secreted (RANTES) was enhanced by direct co-culture compared to indirect co-culture using Transwell. Production ratio of RANTES in OA was significantly higher than non-arthritic samples. CD3 positive mononuclear cells and chondrocyte-like cells were found in SF. Chondrocyte–T cell contact was more adhesive in OA samples. These results showed the production of MMPs and RANTES was enhanced by the interaction and that chondrocyte–T cell contact was possible in vivo.     

兔前交叉韧带切断骨关节炎模型软骨MMP-1 MMP-3及诱导型一氧化氮合酶表达的研究

目的观察兔前交叉韧带切断（ACLT）骨关节炎（OA）模型软骨中不同造模时期基质金属蛋白酶（MMP）-1、3及诱导型一氧化氮合酶（iNOS）的表达，为该模型用于OA防治提供理论依据。方法36只大白兔，随机选择27只，每只均行右侧膝关节腔切开前交叉韧带切断术（ACLT），另9只单纯行右侧膝关节腔切开术作为假手术对照组。术后4、8及12周随机处死实验组大白兔各9只及假手术对照组大白兔3只。对比各组大白兔股骨髁关节软骨的大体改变，用反转录-聚合酶链反应及免疫组织化学的方法检测MMP-1、3及iNOS在软骨中的mRNA和蛋白的表达结果。结果ACLT造模术后4周即开始出现软骨早期退变表现，造模8周软骨退变加重，造模12周出现OA晚期软骨退变表现，不同造模时期的大体评分差异有统计学意义，造模4周MMP-1、3及iNOS表达量明显高于对照组，随着造模时间延长MMP-1、3及iNOS表达量进一步升高，MMP-1、3及iNOS在退变软骨的不同阶段表达分布有各自的特点。结论ACLT模型能够表现OA软骨降解退变的全过程，适宜用于OA防治研究，MMP-1、3及iNOS的高表达在OA软骨退变的病理过程中起着重要的作用，随着造模时间延长、软骨退变的加重，表达量持续升高，适合作为OA防治研究中的评价指标。     

成纤维细胞激活蛋白和基质金属蛋白酶-1在小鼠肝纤维化组织中的表达及其相关性

目的探讨成纤维细胞激活蛋白(FAP)和基质金属蛋白酶-1(MMP-1)在小鼠肝纤维化组织中的表达和两者之间的关系,及其在肝纤维化形成过程中的作用机制。方法建立四氯化碳诱导的小鼠实验性肝纤维化模型,HE染色和Masson染色法判定肝纤维化程度,采用免疫组织化学染色法测定FAP、MMP-1在肝纤维化过程中的时序动态表达。结果模型组中FAP和MMP-1的表达部位和范围都一致,阳性表达主要分布于门管区内血管壁周围及纤维间隔内的成纤维细胞的胞浆中。线性相关分析显示FAP和MMP-1与小鼠肝纤维化程度呈正相关(r分别为0.672,0.684;P0.0001);且FAP与MMP-1的表达也呈正相关(r为0.828;P0.0001)。结论 FAP和MMP-1可能协同作用参与降解细胞外基质和促进肝星形细胞的活化,从而在肝纤维化的发生发展中起作用。     

Twist1,MMP-2和MMP-9在结直肠癌组织中的表达及意义

目的:研究Twist1、MMP-2和MMP-9蛋白在结直肠癌组织中的表达及其相互关系.方法:建立组织微阵列平台,应用免疫组织化学方法检测92例结直肠癌组织Twist1、MMP-2和MMP-9蛋白的表达情况.结果:结直肠癌中Twist1的表达率为64.1%,MMP-2和MMP-9阳性率分别为66.3%和67.4%;Twi...     

开角型青光眼患者房水中MMP-3、TIMP-1的表达及意义

目的检测开角型青光眼患者房水中基质金属蛋白酶（MMP）-3及金属蛋白酶组织抑制因子（TIMP）-1的表达,并探讨其意义。方法采用酶联免疫反应（ELISA）双抗体夹心法检测开角型青光眼（A组）和年龄相关性白内障（B组）患者房水中MMP-3和TIMP-1的水平。结果A、B两组MMP-3水平分别为（68.0310&#177;4.1400）、（69.7941&#177;2.1315）μmol/L;TIMP-1水平分别为（6.7604&#177;0.6632）、（3.1617&#177;0.1603）μmol/L,MMP-3/TIMP-1分别为9.9017&#177;0.3447、22.0592&#177;1.0172,以上指标两组相比,P均〈0.05。结论开角型青光眼患者房水中MMP-3表达,TIMP-1表达升高,二者共同促进细胞外基质的异常堆积从而参与了开角型青光眼的发病机制。     

Simvastatin reduces MMP-3 level in interleukin 1beta stimulated human chondrocyte culture

OBJECTIVES: Matrix metalloproteinases (MMPs) produced by chondrocytes play a role in the development of cartilage degradation in joint diseases. Moreover, inhibition of MMP secretion by macrophages accumulating in arteriosclerotic plaques would account for the plaque stabilising activity of statins in cardiovascular patients. Recently, simvastatin has been shown to inhibit both developing and established collagen induced arthritis in a murine model. We thus decided to investigate the effect of simvastatin on the production of MMP-3 from cultured interleukin (IL)1 stimulated human chondrocytes. METHODS: Cells from human cartilage, obtained from eight subjects with osteoarthritis undergoing surgery for total hip prostheses, were cultured in the presence of different concentrations of simvastatin (5, 10, and 50 micromol/l) with and without IL1beta (5 ng/ml). MMP-3 level was measured in the culture medium after 48 h of incubation. RESULTS: IL1beta stimulation of chondrocytes increased MMP-3 concentration in the cultures (from 0.69 (0.09) to 1.94 (0.12) ng/microg protein). Incubation with simvastatin was associated with a dose dependent reduction in MMP-3 increase, both in the presence (-15%, -17%, and -26% with 5, 10, and 50 micromol/l, respectively) and in the absence (-32% with 50 micromol/l) of IL1beta. The inhibiting effect of simvastatin was completely reversed by the addition of mevalonate (100 micromol/l) or farnesol (10 micromol/l). CONCLUSIONS: Our data show that simvastatin, by blocking HMGCoA-reductase and interfering in the prenylation processes, is able to inhibit MMP-3 production from cultured human chondrocytes that have been either unstimulated or stimulated with IL1beta, thus suggesting a possible additional mechanism for statins in counteracting chronic joint disease related cartilage damage.     

Differential expression of gelatinase B (MMP-9) and stromelysin-1 (MMP-3) by rheumatoid synovial cells in vitro and in vivo

Summary Primary cultures of adherent rheumatoid synovial cells (ASC) are comprised of variable proportions of fibroblasts, macrophages and stellate cells (activated fibroblasts). These cultures were shown to produce the metalloproteinases stromelysin-1 (MMP-3), gelatinase A (MMP-2) and gelatinase B (MMP-9) by Western blotting and zymography techniques. Immunolocalisation studies showed that MMP-3 was mainly produced by the fibroblastic cells whereas MMP-9 was restricted to macrophages (CD68 positive). Subcultured synovial fibroblasts, devoid of macrophages, did not produce MMP-9 as judged by zymography and immunolocalisation; but when stimulated with phorbol myristate acetate and interleukin-1 both MMP-9 and MMP-3 were co-expressed. These activated fibroblasts assumed a dendritic or stellate morphology, which in localisation studies was usually associated with enhanced enzyme production. Immunolocalisation studies of rheumatoid synovial tissue showed that relatively few cells were positive for MMP-3 and MMP-9. Localisation of MMP-9 corresponded to a proportion of macrophages positive for the CD68 marker throughout the synovial tissue. MMP-3 localisation was not associated with the macrophage marker, but was observed in both the synovial lining layer and deeper stromal locations. Widespread distribution of both enzymes was not observed in fresh tissues, but this increased in tissues subjected to short-term explant cultures. Thus, both in vitro and in vivo studies indicated that synovial fibroblasts or B-cells are effective producers of MMP-3 whereas macrophages elaborate MMP-9, observations that demonstrated different metalloproteinase phenotypes under similar environmental conditions.     

Expression levels and association of gelatinases MMP-2 and MMP-9 and collagenases MMP-1 and MMP-13 with VEGF in synovial fluid of patients with arthritis

This study was performed to provide evidence, albeit indirectly, as to which matrix metalloproteinases (MMPs), among the gelatinases MMP-2 and MMP-9 and the collagenases MMP-1 and MMP-13, play a more proactive role in the angiogenic process in arthritic joint. Joint fluid was collected from 33 patients with rhuematoid arthritis (RA) and osteoarthritis (OA), and protein (MMPs and vascular endothelial growth factor (VEGF)) levels were measured by ELISA, and the association of MMPs with VEGF was evaluated in joint fluid of patients with RA or OA. The levels of collagenases (total MMP-1 and total MMP-13) and gelatinases (total MMP-2 and total MMP-9) in RA joint fluid were significantly higher than those in OA fluid. Total MMP-9 levels were significantly associated with VEGF levels in RA fluids, but not in OA fluid, while total MMP-13 levels were strongly associated with VEGF levels in both RA and OA fluid. However, total MMP-2 and total MMP-1 levels were not associated with VEGF levels in either RA or OA joint fluid. Our results indirectly suggest that in RA and OA, MMP-9 and MMP-13 may play a more important role in angiogenesis than MMP-2 and MMP-1.     

MMP－2、MMP－3、TIMP－1及氯沙坦与氨氯地平在老年前期自发性高血压大鼠肾硬化中的作用及其机理探讨

目的 探讨MMPs/TIMPs与高血压性肾损害的关系。方法 将SHR随机分为3组,每组10只。氨氯地平10mg·kg~(-1)·d~(-1)或氯沙坦30 mg·kg~(-1)·d~(-1)治疗3个月,为实验组;以模型SHR为空白对照组。同时以10只WKY大鼠为健康对照组。测量治疗前、后血压变化,血尿素氮、肌酐和24 h尿蛋白定量,观察Ⅳ型胶原(COⅣ)、层连蛋白(LM)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-3(MMP-3)、基质金属蛋白酶抑制剂-1(TIMP-1)在肾组织中的表达。结果 ①SHR血压显著增高;肾功能明显受损;肾组织病理改变显著。肾小球细胞外基质(ECM)成分(如:COⅣ、LM)及TIMP-1表达增多;MMP-3、MMP-2表达减少;②治疗后,血压明显降低;肾功能损害减缓;肾组织病理改变减轻。肾组织中COⅣ、LM、TIMP-1表达减少;MMP-3、MMP-2表达增多。结论 氯沙坦与氨氯地平能延缓高血压性肾损害,改善已失调的MMPs/TIMPs比例;减少ECM在肾小球中的聚积,具有延缓高血压性肾硬化的作用。