雷公藤多苷对胶原诱导性关节炎大鼠中高迁移率族蛋白B1影响的研究

目的 探讨高迁移率族蛋白B1(HMGB1)在类风湿关节炎(RA)治疗中的作用及雷公藤多苷治疗RA的可能机制.方法 建立胶原诱导性关节炎(CIA)大鼠模型建立后,将造模成功的大鼠随机分为模型组、甲氨蝶呤治疗组、雷公藤多苷治疗组、雷公藤多苷和甲氨蝶呤联合治疗组进行药物干预,4周后取材,采用免疫组织化学法检测HMGB1在正常组、模型组和各治疗组大鼠的滑膜与关节中的表达,采用酶联免疫吸附试验(ELISA)法检测HMGB1在正常组、模型组和各治疗组大鼠血清中的含量.统计方法采用方差分析.结果 HMGB1在模型组大鼠滑膜、关节中的蛋白表达以及血清中的含量[(23.8±2.2)ng/ml]明显高于正常组[(7.4±1.6)ng/ml](P＜0.01),3个治疗组大鼠滑膜、关节中HMGB1的表达和血清中HMGB1的含量[分别为(133±3.1),(17.4±4.9),(11.7±1.5)ng/ml]显著低于模型组(P＜0.01),但又高于正常组(P＜0.05).结论 HMGB1参与了CIA滑膜增生、软骨和骨质破坏的病理过程;甲氨蝶呤与雷公藤多苷治疗RA滑膜炎和骨质破坏的分子机制与其降低HMGB1的表达相关.     

脂肪因子脂联素及其受体在类风湿关节炎炎性关节中高表达

目的 研究脂肪因子脂联素(AD)及其受体1,2(AdipoR1,R2)在类风湿关节炎(RA)患者关节液和滑膜组织中的表达.方法 酶联免疫吸附试验(ELISA)检测23例RA和23例骨关节炎(OA)患者关节液中AD水平,实时荧光定量聚合酶链反应(PCR)和Western-blot法分析AD,AdipoR1,AdipoR2在10例RA和10例OA患者滑膜组织中的表达.结果 RA关节液中AD含量约高于OA的2倍(P<0.05),RA滑膜组织中AD、AdipoR1 mRNA表达明显高于在OA中的表达(P<0.05),但两者AdipoR2表达差异无统计学意义.RA滑膜组织中AD和AdipoR1蛋白表达明显高于在OA中的表达,且AdipoR1表达明显高于AdipoR2(P<0.05);AdipoR2蛋白表达在RA和OA中差异无统计学意义.结论 RA炎性关节液和滑膜组织中AD和AdipoR1高表达,可能参与RA病理过程.     

细胞因子在类风湿关节炎患者肺间质病变中的意义

目的 探讨类风湿关节炎(RA)继发间质性肺疾病(RA-ILD)患者血清基质金属蛋白酶-9(MMP-9)、基质金属蛋白酶组织抑制因子(TIMP-1)和转化生长因子β,(TGF-β1)的变化及其意义.方法 RA组29例,RA-ILD组28例(其中早期RA-ILD组16例,中晚期RA-ILD组12例),对照组为健康体检人员29名.采用ELASA法检测血清中MMP-9、TIMP-1和TGF-β1水平.计量资料用-x±s表示,多组间比较采用方差分析,两组间比较采用t检验,关节分级资料采用卡方检验.结果 RA组和RA-ILD组血清TIMP-1水平[(645±220)和(536±188)μg/L]明显高于对照组[(392±92)μg/L],RA-ILD组血清TGF-β1.水平[(13.1±10.0)μg/L]明显高于RA组和对照组[(3.9±2.9)和(2.4±1.7)μg/L],RA组与RA-ILD组血清TIMP-1水平、RA组与对照组血清TGF-β1的水平无明显差别.各组血清MMP-9水平和MMP-9/TIMP-1均无明显差别.中晚期RA-ILD组血清TIMP-1水平[(690±110)μg/L]明显高于早期RA-ILD组[(420±147)μg/L],中晚期RA-ILD组血清TGF-β1水平[(17.9±8.2)μg/L]明显高于早期RA-ILD组[(9.5±9.9)μg/L].中晚期RA-ILD组血清MMP-9/TIMP-1(0.9±0.1)明显低于早期RA-ILD组(1.2±0.4).早期RA-ILD组血清MMP-9水平[(537±309)μg/L]与中晚期RA-ILD组[(595±110)μg/L]无明显差别.结论 血清TGF-β1可以作为RA患者ILD的诊断标志,且在一定程度上可反映RA-ILD肺部病变的严重程度.MMP-9/TIMP-1下降能反映肺部病变的严重程度.血清MMP-9水平不能作为RA-ILD的诊断指标及ILD肺部病变严重程度的判定指标.     

类风湿关节炎患者关节滑膜滑液中结缔组织生长因子表达及其意义研究

目的检测结缔组织生长因子(connectivetissue growth factor,CTGF)在类风湿关节炎(rheumatoid arthritis,RA)患者关节滑液及滑膜中的表达,探索CTGF在RA关节病变中可能发挥的作用。方法采用酶联免疫法(enzyme-linked imm-unosorbent assay,ELISA)方法检测中国医科大学附属第一医院2013年1月至12月收治的10例RA患者和8例骨关节炎(osteoar thritis,OA)患者关节滑液CTGF水平;免疫组化法检测3例RA患者和2例OA患者关节滑膜CTGF的表达;Real-time PCR检测向体外培养的成纤维样滑膜细胞(FLS)中加入外源性肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)后对CTGF表达的影响。结果 RA组的关节滑液CTGF水平高于OA组[(21.00±10.20)μg/L对(8.53±7.71)μg/L)],差异有统计学意义(P0.05);CTGF在RA滑膜组织主要表达于滑膜衬里细胞及衬里下层细胞,而OA滑膜组织仅少量CTGF表达;TNF-α可以上调FLS表达CTGF。结论 RA患者滑液CTGF水平升高,滑膜组织CTGF高表达,提示CTGF可能参与了RA关节炎症的发生发展。TNF-α上调FLS的CTGF表达,这可能是导致RA关节病变的重要因素之一。     

精氨酸升压素对大鼠心肌成纤维细胞诱导型一氧化氮合酶-一氧化氮系统活性的影响

目的　研究精氨酸升压素 (AVP)对大鼠心肌成纤维细胞诱导型一氧化氮合酶 (iNOS) 一氧化氮 (NO)系统活性的影响。方法　胰酶消化法分离培养Sprague Dawley仔鼠心肌成纤维细胞 ,采用硝酸还原酶法、蛋白质印迹和逆转录 聚合酶链式反应观察AVP对心肌成纤维细胞的NO含量、iNOS蛋白水平和iNOSmRNA表达的影响。结果　 (1)不同浓度AVP干预下 ,心肌成纤维细胞的NO含量、iNOS蛋白水平和iNOSmRNA表达都随AVP浓度的增高而增加。其中 10 -7mol/LAVP组和 10 -6mol/LAVP组的NO含量 [(6 9 0 5± 5 5 6 ) μmol/L和 (6 2 86± 6 0 5 ) μmol/L]、iNOS蛋白水平 (0 73± 0 0 6和 0 6 4± 0 0 5 )和iNOSmRNA表达 (0 70± 0 0 3和 0 6 6± 0 0 6 )都显著高于对照组 [(2 9 34± 5 34)μmol/L ,0 2 0± 0 0 5 ,0 2 5± 0 0 4 ]、10 -9mol/LAVP组 [(31 79± 6 5 9) μmol/L ,0 2 4± 0 0 7,0 30±0 0 6 ]和 10 -8mol/LAVP组 [(36 87± 7 89) μmol/L ,0 30± 0 0 9,0 31± 0 0 3]。但 10 -6mol/LAVP组的NO含量、iNOS蛋白水平和iNOSmRNA表达都低于 10 -7mol/LAVP组。 (2 ) 10 -7mol/LAVP干预下CFs的NO含量、iNOS蛋白水平和iNOSmRNA表达都随培养时间的延长而增加。其中 2 4h组和 36h组的NO含量 [(6 5     

α1抗胰蛋白酶在强直性脊柱炎患者滑膜组织中表达的研究

目的 观察α1抗胰蛋白酶(ATA1)在强直性脊柱炎(AS)滑膜组织中的表达分布,并探讨编码该蛋白的基因对AS的遗传效应.方法 选用8例AS,9例类风湿关节炎(RA)和9例骨关节炎患者关节滑膜样本,用蛋白免疫印迹定量分析ATA1的表达水平,用免疫组织化学法定位ATA1在滑膜组织中表达分布.分别选用32例AS、RA、骨关节炎滑液样本分析ATAl在滑液中的表达水平.选用56例AS血液样本,260例RA和160名健康者样本作对照,用水解探针法对标签单核苷酸多态性位点(SNP)(rs2753934,rs2749531和rs6575424)进行基因分型.采用单因素方差分析、LSD检验和x2检验进行统计分析.结果 与RA、骨关节炎滑膜比较,ATA1在AS滑膜中呈高表达,表达率为100％.同样,ATA1在AS滑液中表达量(1.6±0.6)也高于RA(1.4±0.5)和骨关节炎(1.2±0.5)的表达量(P＜0.05).基因分型发现ATAl SNP与AS和RA无明显关联(P＞0.05).单体型分析没有检测到与AS相关的或与RA相关的单体型.结论 ATA1在AS滑膜组织中高表达,提示ATA1及关节滑膜在AS发病过程中起重要作用.     

蛋白激酶C-胞外信号调节激酶1/2信号通路在尼古丁诱导的血管内皮细胞表达纤维溶解酶原激活物抑制物-1中的作用

目的 探讨蛋白激酶C(PKC)-胞外信号调节激酶(ERK)1/2信号通路在尼古丁诱导人脐静脉内皮细胞(HUVECs)表达纤维溶解酶原激活物抑制物-1(PAI-1)中的作用.方法 体外培养HUVECs,采用不同实验条件尼古丁进行干预,ELISA法测定细胞上清液中PAI-1的浓度,观察尼古丁作用的最佳浓度和时间.进一步分别用PKC的抑制剂星型胞菌素staurosporine (STS)和ERK的抑制剂PD98059干预HUVECs,观察PKC或ERK被阻断后对尼古丁诱导的HUVECs 表达PAI-1的影响,ELISA测定各组细胞上清液中PAI-1蛋白的表达水平,RT-PCR检测各组细胞PAI-1 mRNA的表达.结果 100 μmol/L尼古丁组PAI-1蛋白水平[(22.6 ± 1.1)μg/L]明显高于对照组[(14.2± 2.8)μg/L,q=5.64,P＜0.05];以100 μmol/L 尼古丁分别与HUVECs孵育0、4、6、8、12及24 h,各组PAI-1蛋白表达呈时间依赖性升高,并在12 h达到高峰(F=32.063,P＜0.05);尼古丁组PAI-1 mRNA及蛋白含量[(1.32±0.20),(21.08±0.83)μg/L]明显高于对照组[(0.73±0.10),(13.39± 0.93)μg/L,q=8.43、11.97,均P＜0.05];尼古丁+STS组PAI-1 mRNA及蛋白含量[(1.07±0.10),(16.19±2.15)μg/L]较尼古丁组降低(q=5.61、7.61,均P＜0.05),但仍高于对照组(q=7.84、4.36,均P＜0.05);尼古丁+ PD98059组PAI-1 mRNA及蛋白表达[(1.12±0.11),(17.52±1.72)μg/L]低于尼古丁组(q=4.68、5.54,均P＜0.05),仍高于对照组(q=8.77、6.43,均P＜0.05).结论 PKC-ERK1/2信号通路在尼古丁诱导的血管内皮细胞PAI-1表达上调中发挥一定作用.

Abstract：

Objective To explore the role of protein kinase C (PKC)- extracellular signal-regulated kinase (ERK)1/2 signal pathway in the process of plasminogen activator inhibitor-1(PAI-1) protein and mRNA expression in cultured human umbilical vein endothelial cells(HUVECs) induced by nicotine. Methods HUVECs were cultured to examine the effect of nicotine on the expression of secreting PAI-1 in HUVECs on different experimental conditions. The expression of PAI-1 protein was measured by ELISA. PKC inhibitor staurosporine (STS)and ERK1/2 inhibitor PD98059 were used to detect PKC or ERK1/2 function on the expression of PAI-1 in HUVECs induced by nicotine. The PAI-1 mRNA expression was determined by RT-PCR. Results The expression level of PAI-1 protein in 100 μmol/L nicotine treated group [(22.6±1.1) μg/L] increased significantly compared to the control group [(14.2±2.8) μg/L; q=5.64,P<0.05]. After stimulation with 100 μmol/L nicotine for 0,4,6,8,12 and 24 h, the levels of PAI-1 protein increased over time and reached the peak at 12 h (F=32.063,P<0.05).The PAI-1 mRNA and protein expression in nicotine treated group [(1.32±0.20), (21.08 ± 0.83) μg/L] increased significantly compared to the control group [(0.73±0.10), (13.39±0.93) μg/L; q=8.43,11.97,all P<0.05].Compared with nicotine treated group , the PAI-1 mRNA and protein expression in nicotine and STS treated group [(1.07±0.10),(16.19±2.15) μg/L] decreased significantly(q=5.61,7.61, all P<0.05), but still higher than the control group (q=7.84,4.36, all P<0.05). In nicotine and PD98059 treated group, the PAI-1 mRNA and protein expression [(1.12±0.11),(17.52±1.72) μg/L] decreased significantly compared to the nicotine treated group(q= 4.68,5.54, all P<0.05), still higher than the control group (q=8.77,6.43, all P<0.05). Conclusion PKC-ERK1/2 signal pathway may play a partial role in the up-regulation of PAI-1 induced by nicotine in HUVECs.     

IL-18BP和IL-4共表达腺病毒基因对关节炎小鼠滑膜组织COX-2和iNOS表达的作用

目的 本研究构建了共表达小鼠白细胞介素-18结合蛋白(IL-18BP)和IL-4的重组腺病毒载体(Ad mIL-18BP/mIL-4),将其用于胶原诱导型关节炎(CIA)小鼠局部关节基因治疗,以探讨对小鼠滑膜组织环氧化酶-2(COX-2)和诱导型一氧化氮合酶(iNOS)及其诱导产物前列腺素E2(PGE2)、一氧化氮(NO)表达水平的影响.方法 采用雄性的DBA-1/BOM小鼠,建立CIA的小鼠模型,经共表达小鼠IL-18BP和IL-4的重组腺病毒载体局部关节基因治疗后,分别采用半定量反转录一聚合酶链反应(RT-PCR)法和Westem印迹法检测滑膜组织COX-2、iNOS mRNA的表达水平和蛋白表达水平,用竞争ELISA和硝酸酶还原法检测关节滑膜液PGE2和NO的含量,并设AdLacZ组和磷酸盐缓冲液(PBS)组分别为病毒对照组和组织对照组.结果 AdmIL-18BP/mIL-4治疗组滑膜组织COX-2、iNOS mRNA表达水平明显低于AdLacZ对照组(0.15 vs 0.42,P<0.01;0.05 vs 0.77,P<0.01)和PBS对照组(0.15 vs 0.65,P<0.01;0.05 vs 0.64,P<0.01);AdmIL-18BP/mIL-4治疗组滑膜组织COX-2、iNOS的蛋白表达水平明显低于AdLacZ对照组(0.08 VS 0.92,P<0.01;0.11 vs 1.00,P<0.01)及PBS对照组(0.08 vs 0.77,P<0.01;0.11 vs 0.84,P<0.01);AdmIL-18BP/mIL-4治疗组关节滑膜液中PGE2及NO水平明显低于AdLacZ对照组[(0.68±0.06)vs(2.58±0.21)ng/ml,P<0.01;(23.4±2.5)vs(60.0±11.3) μmol/L,P<0.01]和PBS对照组[(0.68±0.06)vs(2.57±0.20)ng/mL,P<0.01;(23.4±2.5)vs(60.3±13.4)μmol/L,P<0.01].结论 共表达小鼠IL-18BP和IL-4的重组腺病毒基因治疗能明显下调COX-2和iNOS的表达水平,减少其诱导产物PGE2、NO的合成,这将为RA的基因治疗提供新的治疗策略.     

脱氧雪腐镰刀菌烯醇致新西兰家兔关节损伤的实验观察

目的 观察脱氧雪腐镰刀菌烯醇(DON)对新西兰家兔膝关节软骨和滑膜的损伤.方法 将15只成年雄性新西兰家兔随机分为3组,对照组正常饲养,高、低剂量组分别投予0.10、0.05 mg/kg剂量的DON,经耳缘静脉注射隔日染毒.20 d后处死家兔,取双侧膝关节,进行常规病理检查,测定关节冲洗液白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)、一氧化氮(NO)水平.结果 光镜下,可见低、高剂量组家兔膝关节软骨细胞变性、坏死.滑膜组织炎细胞浸润.关节冲洗液IL-1β、TNF-α、NO,组间比较差异有统计学意义(F值分别为19.396、18.195、22.136,P＜0.05);与对照组[(0.113±0.043)、(0.138±0.087)μg/L、(23.56±9.35)μmol/L]比较,高剂量组[(0.451±0.091)、(0.575±0.122)μg/L,(70.27±11.53)μmol/L]和低剂量组[(0.295±0.107)、(0.387±0.131)μg/L,(45.32±12.24)μmol/L]明显增高(P＜0.05),且高剂量组高于低剂量组(P＜0.05).结论 DON可导致家兔关节软骨和滑膜损伤,其损伤类似于人骨关节炎病变.DON可能是骨关节炎的病因之一.     

透明质酸钠对兔骨关节炎滑膜iNOS的表达及滑液一氧化氮含量的影响

目的观察透明质酸钠(SH)对兔骨关节炎模型滑膜诱导型NO合酶(iNOS)的表达及滑液中NO含量的影响。方法16只大耳白兔行单膝前交叉韧带切断术,术后5周将动物随机分为实验组和对照组,实验组关节腔注射1％SH 0．3 ml,每周1次,连续5周；对照组则注射等量生理盐水。术后10周处死动物,采用反转录-聚合酶链反应(RT-PCR)方法检测滑膜iNOS的mRNA表达；采用硝酸还原酶法测定滑液NO的含量。结果实验组滑膜iNOS mRNA的表达水平(0．47±0．09)较对照组(0．65±0．12)显著降低(P＜0．01)。实验组滑液中NO的含量[(134±12)→μmol/L]与对照组[(152±16)μmol/L]比较明显下降(P＜0．05)。结论SH对兔骨关节炎关节滑液中NO的含量有降低作用,这可能与SH抑制滑膜iNOS的表达有关。     

类风湿关节炎活动期滑液中葡萄糖6-磷酸异构酶水平及与抗环瓜氨酸肽抗体的关系

目的 检测葡萄糖-6-磷酸异构酶(GPI)在类风湿关节炎(RA)活动期膝关节滑液中的含量,并探讨滑液中GPI与抗环瓜氨酸肽(CCP)抗体的关系.方法 用酶联免疫吸附法(ELISA)检测22例RA活动期患者和37例骨关节炎活动期患者滑液中GPI和抗CCP抗体的浓度.组间比较采用t检验,相关性分析采用Spearman相关分析.结果 RA活动期患者膝关节滑液中的GPI浓度高于骨关节炎活动期患者[(9.6±8.4)和(0.9±1.8)μg/ml],差异有统计学意义(P＜0.01);RA活动期患者与骨关节炎活动期患者滑液的抗CCP抗体浓度分别为(14.61±18.64)和(1.42:±0.09) U/ml,差异有统计学意义(P＜0.01);RA活动期患者膝关节滑液中GPI含量和抗CCP抗体浓度呈正相关(r=0.447,P=0.037).结论 GPI在RA活动期患者滑液中高表达,可能与RA发生发展过程中慢性滑膜炎、骨质破坏有关.GPI与抗CCP抗体呈正相关,两者可作为RA实验室诊断的参考指标.     

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.     

Distinct expression of mast cell tryptase and protease activated receptor-2 in synovia of rheumatoid arthritis and osteoarthritis

The objective of this study is to examine the differential expression of mast cell tryptase and its receptor, protease-activated receptor-2 (PAR-2), in the synovium and synovial fluid of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Biochemical and immunohistochemical analyses were performed to determine whether the trypsin-like protease in the synovium is identical to mast cell tryptase. The effects of mast cell tryptase on the proliferation of synovial fibroblast-like cells (SFCs) and the release of IL-8 thereof were evaluated by the [³H]-thymidine incorporation and ELISA, respectively. The trypsin-like protease in the synovium of RA patients was identical to human mast cell tryptase, which was composed of two subunits: 33 and 34 kDa. The 33- and 34-kDa proteins are different glycosylated forms of the 31-kDa protein, which was unglycosylated. Mast cell tryptase activity in RA synovial fluid was significantly higher than that in OA synovial fluid, while their activities and expression in the synovium were similar. Expression of PAR-2 mRNA in the synovium was higher in RA than in OA. Mast cell tryptase containing the unglycosylated 31-kDa subunit was the predominant form in synovial fluid. RA patients had higher amounts of this subunit in their synovial fluid than OA patients. Mast cell tryptase and PAR-2 activating peptide stimulated the proliferation of SFCs and release of IL-8 from these cells. Mast cell tryptase secretion into RA synovial fluid is higher than OA synovial fluid. Mast cell tryptase in synovial fluid stimulates the proliferation of SFCs and the release of pro-inflammatory cytokines via PAR-2, which may contribute to exacerbation of synovitis in RA.     

Comparative analysis of cathepsin l,cathepsin d,and collagenase messenger rna expression in synovial tissues of patients with rheumatoid arthritis and osteoarthritis,by in situ hybridization

Objective. To compare the expression of cathepsin L, cathepsin D, and collagenase messenger RNA (mRNA) in synovial specimens from patients with rheumatoid arthritis (RA) and osteoarthritis (OA). Methods. The expression of cathepsins L and D as well as collagenase mRNA in synovial tissues from 8 patients with RA, 6 patients with OA, and 2 patients with noninflamed joints was evaluated using in situ hybridization with digoxigenin-labeled RNA probes. Results. Both RA and OA synovial tissue expressed cathepsins L and D as well as collagenase mRNA. The expression of the cathepsins was markedly higher in interstitial regions and, to some extent, in perivascular infiltrates of RA synovial tissue compared with OA specimens. Conclusion. Cathepsins L and D mRNA are expressed differently in RA and OA synovial tissues, supporting the concept that these enzymes may contribute to the influx of mononuclear cells into RA synovium. Moreover, the data reveal that the expression of collagenase and cathepsins in RA and OA synovial lining is otherwise largely similar, and suggest that the adhesion of synovial cells to cartilage mediates the invasive destructive process in RA.     

Increased chromogranin A levels indicate sympathetic hyperactivity in patients with rheumatoid arthritis and systemic lupus erythematosus

OBJECTIVE: Sympathetic hyperactivity is an unfavorable disease consequence in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) due to an increased risk of cardiovascular events. We aimed to identify a serum marker of the sympathetic nervous system, the adrenal chromogranin A (CHGA), in order to study sympathetic hyperactivity in RA and SLE. METHODS: Serum levels of CHGA were measured by radioimmunoassay in healthy subjects and patients with RA and SLE. CHGA immunofluorescence was performed in synovium of patients with RA and controls with osteoarthritis (OA). CHGA levels were measured in plasma, synovial fluid, and synovium superfusate in RA and OA controls. RESULTS: In healthy subjects, systemic CHGA levels correlated positively with age and plasma norepinephrine, indicating the sympathetic origin (p < 0.01). Serum CHGA levels were higher in RA and SLE than in healthy subjects (p < 0.001), which was particularly evident in female patients. Immunofluorescence revealed double-staining of CHGA and elastase-positive neutrophils in the synovium (but not with macrophages, T cells, fibroblasts, B cells, or tyrosine hydroxylase-positive cells). Density of CHGA+ cells was higher in RA synovium compared to OA controls. In OA controls and RA, CHGA levels were similar in plasma and synovial fluid, but levels in synovial tissue superfusate were markedly lower, which indicates that most of the CHGA is of systemic adrenal origin. CONCLUSION: Increased level of CHGA is a good marker of systemic sympathetic hyperactivity.     

Increased levels of circulating intercellular adhesion molecule 1 in the sera of patients with rheumatoid arthritis

Objectdive. We sought to assess whether circulating levels of intercellular adhesion molecule 1 (ICAM-1) in patients with rheumatoid arthritis (RA) are elevated and correlate with clinical measures of disease activity and whether this ICAM-1 originates from the synovium. Methods. Circulating ICAM-1 (cICAM-1) levels were determined by sandwich enzyme-linked immunosorbent assay of serum from 61 RA, 18 osteoarthritis (OA), and 11 inflammatory arthritis (IA) patients. In addition, paired serum and synovial fluid samples were collected from 17 RA, 9 OA, and 4 IA patients. The stability of cICAM-1 was assessed by overnight incubation at 37°C. Finally, the potential degradative effects of synovial fluid proteases were assessed by incubation of recombinant soluble ICAM-1 with patient synovial fluid. Results. RA sera contained significantly greater (P < 0.001) levels of cICAM-1 compared with RA synovial fluid and compared with sera or synovial fluid from the OA and IA patients. Circulating ICAM-1 levels were unaffected by overnight incubation at 37°C and were unaffected by exposure to RA, OA, or IA synovial fluid. Serum levels of cICAM-1 demonstrated a weak, but significant (P < 0.05) correlation with the joint score and erythrocyte sedimentation rate in 25 RA patients treated with nonsteroidal antiinflammatory drugs. Conclusion. The greatest elevations of cICAM-1 were seen in RA patient sera. In both RA and OA, synovial fluid cICAM-1 levels were consistently lower than serum levels, suggesting that cICAM-1 did not originate in the synovium. Because the production of cICAM-1 can be increased by cytokines (e.g., interleukin-1, tumor necrosis factor α), elevated levels of circulating ICAM-1 in RA may be reflective of systemic exposure to elevated cytokine levels.     

Characterization and functional studies of rheumatoid synovial mast cells: Activation by secretagogues,anti-IgE,and a histamine-releasing lymphokine

Microscopic analysis of synovial specimens from 35 patients with rheumatoid arthritis (RA) and 7 patients with osteoarthritis revealed mast cell hyperplasia in perivascular regions, in fibrous interstitial areas, and clustered around the periphery of lymphoid aggregates. Metachromatic staining, immunofluorescence studies, and ultrastructural analysis revealed a single population of connective tissue-type mast cells with surface IgE receptors. Total extractable histamine of synovial tissue was 4.15 ± 2.30 μg/gm (n = 8) for RA synovium and 0.53 ± 0.23 μg/gm (n = 7) for OA synovium. Mast cell secretion was assessed and specific release of histamine from RA synovial mast cells was observed following stimulation with anti-IgE (32.3%), compound 48/80 (40.1%), calcium ionophore A23187 (25.2%), and a partially purified lymphokine with histamine-releasing activity (23.9%).