【作者】 王维山；

【导师】 史晨辉； 董金波； 刘维钢； 王永明；

【作者基本信息】 石河子大学 ， 外科学， 2007， 硕士

【摘要】 目的:通过向兔膝关节腔内注射外源性尿激酶型纤溶酶原激活物,观察其对关节软骨、关节滑膜、关节液的影响,探讨其在骨性关节炎发病中的作用。方法:将健康新西兰大白兔66只按体重、性别随机分为实验组和对照组,实验组24只,对照组42只。实验组按4,8,12W三个不同时间点随机分为3组,每组8只,每只兔在右膝关节内注射外源性尿激酶型纤溶酶原激活物0.2mL(0.4μg);对照组随机分为7组,每组6只:分别为空白对照4W组,未行任何处理;阴性对照4W,8W,12W组,分别在兔右膝关节内注射生理盐水0.2mL;阳性对照4W,8W,12W组,按照Hulth法制作骨性关节炎模型,即切除右膝关节内侧半月板、切断前后叉韧带、切断内侧副韧带。然后分批处死兔取血液、双膝关节软骨、滑膜、关节液进行相关指标的检测。结果:(1)实验组兔右膝关节软骨表面不规则、缺损及结构破坏,软骨细胞数目明显减少、纤维增生,并出现潮线的完整性破坏,关节软骨退变的Mankin’评分明显高于阴性对照组(P<0.01)。电镜下软骨细胞开始溶解,外形不规则,胞膜突起消失,胞浆内细胞器基本完全消失,细胞核开始部分溶解;同时膝关节X线片检查示符合OA影像学病变。(2) uPA诱导的骨性关节炎模型与阳性对照组Hulth法制作的骨性关节炎模型在组织病理变化、影像等方面相似。(3) uPA与基质金属蛋白酶-3在骨性关节炎模型血液和关节液中的含量呈现明显的正相关关系(P<0.01),其含量亦随着软骨降解程度的加重而增加。结论:(1) uPA关节腔内注射法制作的兔膝关节骨性关节炎模型,在病理组织、影像学等方面的改变与人骨性关节炎基本一致,且制造方法简单,制作周期较短,为一种较为理想的骨性关节炎动物模型。(2)uPA作为一种细胞外基质的降解酶系,不但可促使软骨发生降解,而且可激活体内其他细胞因子及细胞外基质降解系统,从而共同促进软骨降解。(3)通过检测血清和关节液中uPA和基质金属蛋白酶-3的含量可辅助诊断骨性关节炎、判断病情和预测预后。更多还原

【Abstract】 Purpose:To investigate the effects of the urokinase type plasminogen activator in the pathogenesis of osteoarthritis by study on the histological change of cartilage、synovium of knee joint and the induction of cytokines in synovial fluid in experimental animals with intra-articular injection of urokinase type plasminogen activator into the one knee joint of rabbits. Methods:The 66 rabbits were divided into experimental animals and control animals according to the weight and the sex, 24 cases for experimental group and 48 cases for control group. Experimental animals were randomly divided into three groups acording to time, 8 cases for each group and were injected intraarticularly with 0.4ug/0.2ml uPA. Control animals were randomly divided into seven groups, 6 cases for each group, included one blank control group which was not given any injection medicine; three negative control groups were injected intraarticularly with 0.2ml normal saline; three positive control groups were transected of the medial collateral ligament, medial meniscectomy, anterior cruciate ligament and posterior cruciate ligament. The rabbits were respectively sacrificed at weeks 4, 8, and 12 week. Histological change of cartilage and synovium of knee joint and the induction of cytokines in synovial fluid in each animals were detected.Results:1. Histological detection and scale of rabbit right kneel articular cartilage in experimental groups under light microscope: the surface of the joint cartilage showed defects, damages, structural breakage, and the number of cartilage cell decreased. Collagen fiber hyperplasia and tidal line of complete break while 4-layer structure of articular cartilage in the blank control group and negative control groups was clear to see, and cartilage cells were arranged in order with complete tidal line. Mankin’s evaluation of the degeneration in articular cartilage in experimental groups and negative control groups: the scores at 4^th ,8^th ,12^th weeks were higher in experimental groups than those in the negative control groups（P<0.01）. 2. The uPA induced OA model was similar with Hulth-OA model in histology of cartilage and imageological findings of knee.3. Imageological findings comformed to OA of human-being,the uPA、MMP-3 contents in the synovial fluid were markedly higher than that of in the negative control groups. There is a significant linear relationship between uPA and MMP-3 in serum and synovial fluid from the experimental groups（P<0.01）, and the contents increased to different extents with increase of cartilage degeneration.Conclusion:1. Experimental knee OA of rabbits with intra-articular injection uPA is in conformity with the appearance of OA in histomorphology and pathematology, and production process is simple, therefore it is a good animal model of experimental OA.2. As one of catabolic enzymes that act on the degradation of extracellular matrix, uPA not only directly degenerates cartilage, but activates other catabolic enzymes of extracellular matrix and correlated cytokines. It maybe plays a key role during the occurrence and development of of OA as one of inducing factor.3. The MMP-3 and uPA levels in serum and synovial fluid of the knees can reflect the degree of synovitis and accumulative cartilage injury as a biomarker, especially accumulative cartilage injury.更多还原