布洛芬与氨基葡萄糖对膝骨关节炎患者滑膜细胞增殖及软骨寡聚基质蛋白表达影响的比较

目的 探讨布洛芬与氨基葡萄糖对膝骨关节炎(OA)滑膜细胞增殖及软骨寡聚基质蛋白(COMP)表达的影响.方法 布洛芬与氨基葡萄糖含药血清培养早期和晚期滑膜细胞,四唑氮化合物/黄嘌呤氧化酶(MTS/PMS)法测定吸光度(A)值,hCOMP定量试剂盒测定COMP含量.采用等方差假设双侧t检验进行统计学处理.结果 通过MTS/PMS法观察确定布洛芬与氨基葡萄糖含药血清培养滑膜细胞观察点在第5～7天;氨基葡萄糖含药血清培养滑膜细胞A值[晚期组(0.054±0.021),早期组(0.777±0.034)]低于正常血清对照组(P＜0.05).布洛芬[晚期组(35.4±1.9),早期组(46.0±2.2)]与氨基葡萄糖含药血清[晚期组(36.6±1.3),早期组(48.8±1.3)]对软骨寡聚基质蛋白的表达也有明显降低作用(P＜0.05).结论 氨基葡萄糖可以抑制体外培养早期和晚期膝OA患者滑膜细胞增殖,而布洛芬与氨基葡萄糖皆可抑制体外培养滑膜细胞分泌COMP.     

柔肝中药对体外培养软骨细胞增殖能力及关节软骨低聚基质蛋白分泌的影响

目的 探讨柔肝中药对体外培养软骨细胞增殖能力及关节软骨低聚基质蛋白（COMP）分泌的影响。方法 采用分阶段酶消化法体外培养兔软骨细胞，以2×10^4／ml密度接种3代内细胞，以Ⅱ型胶原的免疫组织化学研究鉴定细胞。给予家兔临床等效剂量灌胃后，抽取含药血清培养细胞，分别用5％、10％的柔肝方含药血清（分给药后1、3、5h3个时间点）以及正常兔血清、小牛血清干预7d，采用四甲基偶氮唑蓝（MTF）法观察中药含药血清对体外培养的软骨细胞增殖的影响；体外培养人软骨细胞，采用柔肝复方组及单味柔肝药提取物直接添加体外培养体系3d，添加终浓度为10mg／ml。采用酶联免疫吸附试验（ELISA）检测药物对体外培养的人软骨细胞COMP分泌的影响。结果 培养细胞经Ⅱ型胶原的免疫组织化学染色后有阳性表现，在对软骨细胞增殖的影响方面，柔肝方含药血清各组均优于兔血清组和小牛血清组，其中含药血清组的3h时间点总体优于1h及5h时间点（P〈0．05）；在对软骨细胞上清COMP分泌方面，柔肝复方及单方提取物组均有促进细胞分泌COMP的作用（P〈0．05），但两组之间差异无统计学意义（P〉0．05）。结论 柔肝方中药含药血清可以促进体外培养软骨细胞增殖；在药物直接干预的条件下，柔肝复方及单方提取物可以促进软骨细胞分泌COMP。     

复元胶囊对膝骨关节炎模型血清软骨寡聚基质蛋白的影响

目的 观察复元胶囊对实验性膝骨关节炎(OA)模型病理形态及血清中软骨寡聚基质蛋白(COMP)的影响.方法 72只雄性SD大鼠随机分成正常组、模型组、壮骨关节丸组、复元胶囊组,每组18只.除正常组外,其余各组采用Hulth法建立膝OA动物模型.造模后,壮骨关节丸组、复元胶囊组分别给予壮骨关节丸、复元胶囊灌胃,而正常组、模型组给予生理盐水.灌胃后4、8、12 w分别随机选取6只进行X线摄片取材.通过X线摄片、肉眼观察、Mankin评分以及电镜检测评价复元胶囊治疗OA疗效,酶联免疫吸附法检测血清中COMP水平.结果 复元胶囊组膝关节x线积分、Mankin评分在4、8、12 w三个时间点均显著低于模型组(P<0.01),复元胶囊组血清COMP水平也低于模型组(P<0.01).结论 复元胶囊能降低血清软骨代谢标志物COMP水平,延缓OA软骨退变过程.     

骨关节炎兔血清中软骨寡聚基质蛋白和基质金属蛋白酶-3水平

目的 探讨血清软骨寡聚基质蛋白(COMP)与基质金属蛋白酶(MMP)-3检测应用于临床评估骨关节炎(OA)软骨病理改变的可能性.方法 伸直位石膏管型制动16只兔右后膝关节制作OA模型.以造模时间不同分为造模2周、造模6周,左后膝关节未造模故为对照组.X线影像学与病理观察模型关节的变化;评估关节软骨降解程度(OA积分);ELISA检测兔血清COMP、MMP-3水平;分析血清COMP、MMP-3水平与OA积分间的相关性.结果 (1)造模2周影像学变化较造模前不明显;造模6周兔胫骨平台边缘不光滑,关节间隙变窄,表面有毛刺样增生,胫骨平台及股骨内髁外侧可见唇样增生.(2)OA关节病变的形态学观察:造模2周兔关节软骨表面粗糙,表层裂隙;软骨细胞弥漫增多,排列紊乱;OA积分为(4.000±2.204)分.造模6周兔关节软骨表层可见较多裂隙延伸向下深达辐射层;裂隙周围可见脱水固缩坏死的软骨细胞且排列紊乱,部分成簇增生,各层结构不易分辨,有血管翳通过;OA积分为(10.620±1.408)分,与造模2周比,P=0.000.(3)造模2周兔血清COMP[(3.64±0.18)μg/L]、MMP-3[(1.99±0.81)μg/L]水平高于造模前[COMP(3.35±0.20)μg/L,MMP-3(1.61±0.71)μg/L];造模6周兔血清COMP[(3.96±0.44)μg/L]、MMP-3[(3.44±0.91)μg/L]水平高于造模前和造模2周,差异有统计学意义(P值均＜0.05).血清COMP、MMP-3水平与OA积分呈线性相关关系(r值均＞0.710,P值均小于0.05).结论 OA血清中COMP和MMP-3水平对评估OA软骨降解程度具有重要意义.

Abstract：

Objectiye To study the levels of cartilage oligomeric matrixprotein (COMP) and matrix metalloproteinase-3 (MMP-3) in the serum fluid of osteoarthritic rabbit models and their relationships with the severity of pathological changes, so as to investigate their correlation with osteoarthritis(OA). Methods The osteoarthritic animal models were get from immobilizing the right knees of 18 rabbits in full extension using plaster cast. Knee joint pathological changes of 2,6 weeks were examined for pathological severity of OA; ELISA sandwich method was used to measure the levels of COMP and MMP-3 in serum before and after modeling( at 2, 6 weeks respectively); X ray of model keens was also obtained in different period.Correlation analysis was performed to demonstrate the relationship between the levels of COMP, MMP-3 in the serum and the pathological severity of OA. Results ( 1 ) Morphological observations: immobilizing the right knees of rabbits in full extension using plaster cast was a reliable methed for osteoarthritic animal models and the typical histopathologic character was seen; the severity of osteoarthritisgradually increased with time extended. (2) The levels of COMP[(3.64 ±0. 18)μg/L], MMP-3 [(1.99 ±0. 81 ) μg/L]in the serum of 2 weeks osteoarthritic animal models were higher than those before immobilizing with plaster cast [COMP(3.35 ±0. 20) μg/L,MMP-3( 1.61 ±0. 71 ) μg/L]. The levels of COMP[(3.96 ±0. 44) μg/L],MMP-3[(3.44 ±0. 91) μg/L] of 6 weeks were much higher,with a significant difference(P ＜0.05). The levels of COMP, MMP-3 in serum had a linear correlation with the pathological severity of OA (r ＞0. 710,and P ＜ 0. 05 ). Conclusion The levels of COMP and MMP-3 in serum can help to predict and evaluate the progression of OA.     

软骨寡聚基质蛋白对骨关节炎软骨破坏早期诊断价值的研究

目的 探讨软骨寡聚基质蛋白(COMP)对骨关节炎软骨破坏早期诊断价值.方法 兔右后膝伸直位石膏管型固定法制作骨关节炎模型;形态学方法观察造模不同时期关节病理切片;免疫组织化学方法检测软骨内COMP水平;酶联免疫吸附试验(ELISA)方法检测血清COMP水平.应用t检验,Pearson相关性分析.结果 ①伸直位石膏管型制动2周,模型关节呈现早期骨关节炎改变,制动6周呈现典型的中晚期骨关节炎特征;②造模前、模型2周、模型6周血清COMP含量分别是[(3.35±0.20)、(3.64±0.18)、(3.96±0.44)μg/L,P均＜0.05];③未造模、模型2周、模型6周关节软骨内COMP表达强度分别是[(2.7±1.8)%,(5.7±0.7)%,(7.6±0.7)%,P均＜0.05];④模型2周血清COMP水平与模型2周组、模型6周组OA病理评分存在线性相关关系(r均＞0.770,P均＜0.05).结论 骨关节炎血清COMP检测对早期诊断骨关节炎软骨破坏具有重要的意义.

Abstract：

Objective To study the diagnostic value of cartilage oligomeric matrix protein for early cartilage destruction in osteoarthritis and assess its value in the prediction of the disease progression.Methods The osteoarthritis animal models were developed by immobilizing the right knees of 18 rabbits in full extension position using plaster East.Knee joint pathological changes at week 2 and 6 were examined for pathological severity evaluation of osteoarthritis.ELISA sandwich method was used to measure the levels of cartilage oligomeric matrix protein(COMP) in serum before and after modeling(at week 2 and 6 respectively) and immunohistolgy method was used to examine the levels of COMP in knee articular cartilage of osteoarthritis animal models.Correlation analysis was performed to demonstrate the relationship between the levels of COMP in the serum and the pathological severity of osteoarthritis.Pearson's test and t-test were used for correlation analysis.Results ①) Osteoarthritis animal models could be successfully developed by immobilizing the right knees of rabbits in full extension position using plaster east for 2 weeks.Early histopathological changes in the articular cartilage could be observed,At week 6,the typical histopathological characteristics could be seen.②With the extension of modeling time,serum COMP levels persistently increased.The serum COMP levels before modeling,at modeling week 2,week 6 were (3.35±0.20),(3.64±0.18),(3.96±0.44) μg/L respectively,the difference was significant (P＜0.05).③ The level of COMP in the articular cartilage of non-osteoarthritis animal models,models at week 2,week 6 were (2.7±1.8 )％ ,(5.7±0.7)％,(7.6±0.7)％ respectively (P＜0.05 for all).④ The level of COMP in the serum was linearily correlated with the pathological severity of osteoarthritis(r＞0.770 for all,and P＜0.05 for all).Conclusion Levels of COMP in the serum can help to make early diagnosis of osteoarthritis,and elevated COMP level can predict the progression of osteoarthritis.     

骨关节炎患者关节软骨和滑膜金属蛋白酶-1和-3的表达

骨关节炎 (ＯＡ)的病理特征为关节软骨的进行性变性和破坏 ,但其确切的发病机制未明。近年研究提示 ,ＯＡ的这种病理改变与金属蛋白酶 (ＭＭＰｓ)对细胞外基质的降解作用可能有关[1] 。我们检测了ＭＭＰ 1、 3在ＯＡ软骨和滑膜组织中的表达状况 ,以探讨两者在ＯＡ发病中的作用。一、对象与方法1 实验对象 :膝ＯＡ病人均符合Ａｌｔｍａｎ等[2 ] 的ＯＡ诊断标准。正常对照组包括意外受伤截肢患者或意外死亡者。膝ＯＡ患者 14例 ,男 5例 ,女 9例 ,平均年龄 (5 8 2± 11 1)岁 ;正常对照组 11例 ,男 5例 ,女 6例 ,平均年龄 (5 1 3± 9 6 )岁。2 …     

骨关节炎软骨细胞和滑膜细胞金属蛋白酶活性的影响因素

目的 探讨几种细胞因子、生长因子和药物等刺激物对骨关节炎软骨细胞及滑膜细胞金属蛋白酶活性的影响。方法 骨关节炎患者的单层软骨细胞和膜细胞均以白细胞介素-1β、转化生长因子-β1、肿瘤坏死因子-α、双氯芬酸钠、多西环素和地塞松分别处理72h。应用酶谱分析细胞培养上清液中酶活性。结果 单层软骨细胞产生金属蛋白酶-9和-2,而滑膜细胞仅产生金属蛋白酶-2。白细胞介素-1β,肿瘤坏死因子-α和双氯芬酸钠均能显著增强金属蛋白酶-9活性,其中以白细胞介素-1β作用最强,与肿瘤坏死因子-α或双氯芬酸钠有协同作用。多西环素、转化生长因子-β1和地塞米松均能抑制金属蛋白酶-9和-2活性,并与白细胞介素-1β、肿瘤坏死因子-α和地塞米松均能抑制金属蛋白酶-9和-2活性,并与白细胞介素-1β、肿瘤坏死因子-α和双氯芬酸钠起拮抗作用。多西环素为最强的抑制物。结论 白细胞介素-1β和肿瘤坏死因子-α可能为破坏关节软骨的重要细胞因子,转化生长因子-β1则为保护因子,多西环素则可能成为骨关节炎的有效治疗药物。     

软骨寡聚基质蛋白的生物学特征及与血管内成形术后再狭窄的关系

软骨寡聚基质蛋白（cartilage oligomeric matrix protein，COMP）是近年来新发现的一种细胞外基质蛋白，最初在骨科领域被广泛研究。随着研究的不断进展，研究人员发现其作为血管的组成部分，对血管损伤后再狭窄有一定影响。现就COMP的结构、合成、调控、生物学功能以及与血管内成形术后再狭窄的关系进行综述。     

壳聚糖膜对体外培养大鼠软骨细胞增殖活性及分泌细胞外基质的影响

目的 探讨壳聚糖膜对体外培养大鼠软骨细胞增殖活性及分泌细胞外基质的影响.方法 制备壳聚糖膜材料.体外分离培养大鼠膝关节软骨细胞,随机分为6孔板细胞组、种植于壳聚糖膜细胞组、PBS处理组、壳聚糖膜浸出液处理组.通过CCK-8法检测培养不同时间软骨细胞的增殖活性,并通过Real-time PCR法检测不同培养时间软骨细胞分泌Ⅱ型胶原及聚集蛋白聚糖的量.结果 通过Ⅱ型胶原免疫组化染色发现,本实验培养的细胞呈阳性,表明该细胞为软骨细胞;CCK-8检测结果表明,种植于壳聚糖膜细胞组、壳聚糖膜浸出液处理组的软骨细胞增殖活性分别高于6孔板细胞组、PBS处理组(P均＜0.05);Real-time PCR检测结果表明,种植于壳聚糖膜细胞组、壳聚糖膜浸出液处理组的软骨细胞分泌Ⅱ型胶原及聚集蛋白聚糖的量分别高于6孔板细胞组、PBS处理组(P均＜0.05).结论 壳聚糖膜对体外培养大鼠软骨细胞增殖及分泌细胞外基质具有促进作用.     

软骨细胞外基质对生长板软骨构建的影响

观察了各种软骨细胞外基质对骺板软骨体外构建的影响。证实了胶原、蛋白多糖和透明质酸均显著地促进培养软骨细胞的生长,然而各种软骨细胞外基质促进软骨细胞增殖分化的作用并不相同。从组织学可观察到,胶原蛋白明显地促进软骨细胞的肥大化;而蛋白糖和透明质酸抑制成熟期软骨细胞向肥大期转换。     

血清软骨寡聚基质蛋白检测在关节炎患者中的意义研究

目的 探讨血清软骨寡聚基质蛋白(COMP)在各类关节炎患者中的差别及对类风湿关节炎(RA)软骨破坏放射学改变的早期预测价值.方法 采用酶联免疫吸附试验(ELISA)方法测定并对比分析154例各类关节炎患者血清COMP水平的差异,RA患者各项临床指标及2年后关节放射学改良SHARP(vdH-Sharp)指数评分与COMP行相关性分析.结果 与仅有滑膜损害的其他关节炎患者及正常人群比较,RA患者血清COMP显著升高(P＜0.05).骨关节炎(OA)、银屑病关节炎(PSA)患者与正常人群比较血清COMP亦有差异,而与其他滑膜关节炎患者比较差异不大.RA、OA、PSA患者间血清COMP值比较差异无统计学意义.RA患者2年随访发现COMP与患者关节X线的改良SHARP评分前后差值呈正相关(P＜0.01,r=0.848).但与初诊时的抗环瓜氨酸肽(CCP)抗体、类风湿因子(RF)、关节功能、改良SHARP评分均无相关性,与初诊时的红细胞沉降率(ESR)、C反应蛋白(CRP)、晨僵时间、关节肿胀、压痛指数相关(P＜0.05).结论 COMP在各类软骨受侵犯关节炎尤其是RA患者血清中异常增高,提示其可为RA软骨病变早期诊断及判断软骨病变进展、预后和治疗效果的一项理想指标.     

Persistent high serum levels of cartilage oligomeric matrix protein in a subgroup of patients with traumatic knee injury

The objective was to assess whether changes of cartilage oligomeric matrix protein (COMP) serum levels can predict the development of osteoarthritis following traumatic knee injury. Sera and synovial fluids were acquired at surgery (T0) and postoperatively during the first (T1) and second (T2) year from 30 knee-injured patients. COMP levels and anti-COMP autoantibodies were quantified by ELISA. Radiographs and patient questionnaires were used to assess outcomes. At T0, compared with controls (1.6±1.6 μg/ml), the serum COMP concentration was significantly elevated (6.5±2.8 μg/ml) with a tendency to further increase (T0 vs. T1, P=0.076) and subsequently decrease (T1 vs. T2, P=0.074). However, individual variations are observed, e.g. persistently high (8/30) or increasing (T0 to T2, 8/30) serum COMP. Ten of these patients have elevated COMP at T2 that increased from T0. COMP levels in serum and synovial fluid correlated significantly (P=0.012). Interestingly, some patients who revealed increasing serum levels of COMP from T0 to T2 displayed anti-COMP autoantibodies. These data suggest that local immune response could contribute to further joint damage. The subgroup of 10 patients (33%) with elevated and increasing serum COMP levels and in particular the patients with antibodies against cartilage matrix molecules appear at increased risk for developing posttraumatic osteoarthritis. Received: 27 August 1997 / Accepted: 2 March 1998     

Persistent elevation of fibrosis biomarker cartilage oligomeric matrix protein following hepatitis C virus eradication

Serum levels of cartilage oligomeric matrix protein (COMP) has been presented as a biomarker of liver fibrosis in several cross-sectional studies. COMP is also an essential mediator in carcinoma development and has also been associated with hepatocellular carcinoma. We present a prospective analysis of this biomarker in 38 patients with chronic hepatitis C who were subject to eradication therapy with direct acting antivirals. We confirm previous studies associating COMP elevation with liver cirrhosis. We also show how viral levels are correlated with COMP at baseline. In our prospective analysis, we report that successful eradication of hepatitis C results in improvement in liver stiffness and laboratory liver function tests at 1 year follow-up. In contrast, median COMP-levels remain unchanged during the study period. We conclude that the biomarker potential of COMP in the prospective evaluation of liver diseases, remains to be elucidated.     

类风湿关节炎患者血清软骨寡聚基质蛋白水平及意义的研究

目的 探讨检测软骨寡聚基质蛋白(COMP)水平对类风湿关节炎(RA)患者疾病活动性及早期软骨破坏的临床意义.方法 采用酶联免疫吸附试验(ELISA)法,检测94例RA患者及40名健康人血清中COMP的水平.同时测定RA患者的其他实验室及临床指标:类风湿因子(RF)、红细胞沉降率(ESR)、C反应蛋白(CRP)、抗环瓜氨酸肽抗体(抗CCP抗体)、压痛关节数、疼痛关节数、肿胀关节数、晨僵时间及双手X线,分析它们与COMP的相关性.结果 RA患者血清COMP水平(11.3±5.2)U/L明显高于正常对照组的(9.2±1.7)U/L;两组差异有统计学意义(p=0.017).64例RA活动期患者血清COMP水平为(14±6)U/L,30例缓解期患者为(9±4)U/L,组间比较差异有统计学意义(p=0.005).COMP水平与RA思者的ESR、CRP、晨僵时间、压痛关节数、疼痛关节数、肿胀关节数、X,线分期呈正相关(P<0.05),与年龄、病程、RF、抗CCP抗体水平、功能分级无明显关系(p＞0.05).30例RA患者完成2年放射学随访,18例COMP值大于健康者,16例X线有递增现象,2例无递增现象;而12例COMP值正常/低于健康者,有递增5例,无递增7例,两组间差异有统计学意义(p=0.013).结论 RA患者血清中COMP高水平存在提示RA疾病活动性和早期关节软骨破坏.COMP可以作为反映RA疾病急性活动程度与软骨破坏的血清学指标.     

Serum cartilage oligomeric matrix protein (COMP) in rheumatoid arthritis and knee osteoarthritis

The cartilage oligometrix matrix protein (COMP) is a noncollagenous protein, a glycoprotein, the function of which is to bind to type II collagen fibres and stabilise the collagen fibre network in the articular cartilage. In the serum of the normal population the COMP level is 5 g/ml. An increased level of COMP in the synovial fluid was described in the early stage of rheumatoid arthritis (RA), whereas in advanced stages of RA, the level of COMP decreased. In this study we assessed the serum COMP level in patients with RA and knee osteoarthritis (OA) and found a correlation between the serum COMP level and other markers as well as bone mass density (BMD) changes, activity of disease, disease duration and the age of the patients. The blood was collected from 30 RA patients and 30 OA patients who constituted the control group. The serum COMP level was determined using an inhibition enzyme-linked immunosorbent assay (ELISA). The average value of the serum COMP level in RA patients was 10.4±3.6 U/l. There was a correlation between the serum COMP level and the age of RA patients (p<0.005) and disease activity score (DAS) value (p<0.01). According to correlation coefficients, the serum COMP level was independent of stage of disease, number of painful and swollen joints, duration of morning stiffness, disease duration and titre of the Waaler–Rose test. The influence of rheumatoid nodule presence on the serum COMP level was shown (p<0.05). In RA patients with erythrocyte sedimentation rate (ESR) values below 20 mm/h compared with patients with ESR values over 60 mm/h, the serum COMP level was observed to be significantly lower (p<0.05). The average value of COMP in OA patients was 10.4±2.7 U/l. No correlation was found between the serum COMP level and patients age and disease duration. There was a correlation between the serum COMP level and Western Ontario and McMaster Universities (WOMAC) index pain scale for the lower limbs (p<0.005) and T-score value of densitometry examinations (p<0.036) in OA patients. No statistical differences were found between the average serum COMP level in RA and OA patients.     

应用离心管技术以异体微粒软骨脱细胞基质为支架体外构建组织工程软骨

目的将软骨细胞与异体软骨微粒脱细胞基质相结合．构建组织工程软骨。方法分别利用氯化钾、胰蛋白酶和乙二胺四乙酸钠对绵羊关节软骨进行脱细胞，并制成直径0．100—0．154mm的微粒。先分离异体关节软骨细胞并进行体外扩增，再将异体软骨细胞与软骨微粒脱细胞基质混合培养，离心后应用离心管体外培养。结果本脱细胞方法可以使关节软骨细胞完全脱落，且软骨细胞紧紧围绕于异体软骨微粒脱细胞基质四周，生长和分泌功能良好。结论软骨细胞与异体软骨微粒脱细胞基质有良好的生物相容性．可于体外形成软骨样组织。