【作者】 李涧；

【导师】 董启榕；

【作者基本信息】 苏州大学 ， 骨科学， 2010， 硕士

【摘要】 目的:探讨软骨细胞内Toll样受体(Toll-like receptors,TLRs)在骨关节炎(Osteoarthritis,OA)进程中表达的特点,以及受降钙素(Calcitonin,CT)干预后的影响。方法:将75只SD大鼠随机分为三组,手术组与CT干预组各35只,假手术组5只。对手术组与CT干预组的右膝行前交叉韧带切断加内侧半月板切除术,左膝作为空白对照,不予手术。假手术组的右膝只进入关节腔,不处理交叉韧带和半月板。CT干预组动物于术后第二天始行CT15IU/kg隔日皮下注射。前两组大鼠分别于术后7、10、14、21、28、42及56天处死,每组每次处死5只,假手术组动物于术后56天处死并提取标本。对标本进行大体和组织学切片观察。另用Real-time PCR法对手术组和CT干预组动物双膝关节内的软骨标本分别进行TLR2、TLR3、TLR4表达的检测。结果:手术组大鼠造模7天后可见组织学变化;10天后大体观察出现软骨面暗淡,失去光泽;28天后可见骨赘形成;56天后见软骨缺损,软骨下骨外露。Mankin评分由7天的1.0分到28天的6.6分发展到56天的12.2分,与假手术组相比有显著差异。与手术组相比在同一时间点上,CT干预组大鼠的滑膜增生较少;软骨损伤较轻;骨赘形成较小;Mankin评分也较低。手术组膝关节的软骨细胞中TLR2的表达早期增高,晚期明显受到抑制;TLR3的表达与病变严重程度呈正相关性;TLR4表达增高,但不随病程发展而变化。降钙素干预后TLR2表达早期受到抑制,28天后开始增高;TLR3的表达早期也受到抑制,但随着病程的进展表达逐渐的升高;TLR4的表达在早期也同样受到了抑制。结论:采用前交叉韧带切断加内侧半月板切除术可以有效的制作大鼠的OA模型;OA病程中,软骨内有TLRs的表达,TLR2先增高后降低;TLR3与病变严重程度呈正相关性;TLR4高表达但在病程中无明显变化。加用降钙素干预后,尤其是在早期,可明显抑制TLRs的表达,OA的症状也明显改善,在OA早期阶段有一定的治疗作用。更多还原

【Abstract】 Objective: To study the characteristics of the expression of Toll-like receptors ( TLRs) in chondrocyte in the process of Osteoarthritis(OA), and the effect after the intervention of Calcitonin (CT) .Methods: 75 SD rats were randomly divided into three groups, of which Surgery group and CT intervened group both consisted 35 rats, while non-surgery group consisted 5. Surgery Group and CT intervened Group were intervened by cutting off the anterior cruciate ligament and the medial meniscus of their right knees, while the left knees as the control samples. For the non-surgery group, we did not deal with the cruciate ligaments and meniscus , but only exposed the knee joint cavity. CT intervened Group were subcutaneous injected with CT 15IU/kg every other day from the second day of the surgery. The rats of CT intervened Group and Surgery Group were executed and the samples were extracted at 7,10,14,21,28,42, and 56 days after the surgery with 5 rats each time. Non-surgery group rats were executed at 56 days after knee joint exposion. Specimen’s general and histological sections were observed. Both sides of the knee cartilage specimens were carried out within the TLR2、TLR3、TLR4 expression detection by Real-time PCR.Results: The model of Group ACLT+MMx can be seen the histological changes 7 days after surgery; after 10 days the general observation of cartilage surface appears dull; after 28 days the formation of osteophyte were shown; after 56 days the defection of cartilage can be seen with exposion of subchondral bone. It is witnessed the development of Mankin score from 7 days of 1.0 to 28 days to 56 days of 6.6 points to 12.2 points. It were significantly different compared with the non-operated group. The expression of TLR2 of the knee cartilage cells of Group ACLT+MMx increased early and inhibited remarkablely in the later stage. The expression of TLR3 was correlated with the severity of diseases. The expression of TLR4 was high but does not change with the course of OA. After using the intervention of Calcitonin, the early expression of TLR2 was inhibited while 28d later the inhibition stopped and the expression increased. TLR3 expression was curtailed early with no change of the increasing tendency. The expression of TLR4 in early stage was also being inhibited.Conclusion: The resection of anterior cruciate ligament and medial meniscectomy can effectively produce rat OA model. The expression of TLRs in OA process of chondrocyte, of which TLR2 first increased, then decreased, the trend of expression of TLR3 was correlated with the disease severity. The expression of TLR4 was high but does not change with the course of OA. The expression of TLRs was significantly reduced and the symptoms of OA can significantly improve after using the intervention of Calcitonin, especially in the early stage , which have a certain therapeutic effect on OA.更多还原