【作者】 张瑾；

【导师】 杨力；

【作者基本信息】 重庆大学 ， 细胞生物学， 2009， 硕士

【摘要】 在运动中,膝关节内的前交叉韧带是极容易受伤的组织,且损伤后不可自主修复。与此同时,内侧副韧带受损后却可以自我愈合。而MMPs(基质金属蛋白酶)与TIMPs(基质金属蛋白酶抑制剂)在参与调节组织的降解及再生的新陈代谢动态平衡过程中起着极为重要的作用。因此,前交叉韧带及内侧副韧带受损后,基质金属蛋白酶及其天然抑制剂的分子响应机制可能存在差异,这可能是导致两者修复能力不同的原因之一。本研究首先检测前交叉韧带成纤维细胞及内侧副韧带成纤维细胞中上述这两种分子是否有表达。本实验设计了适用于Realtime PCR的引物序列,首先在RT-PCR水平上检测了MMPs及TIMPs家族所有成员在两种成纤维细胞中的表达情况,发现除了MMP-8, 10, 12, 13, 15, 16, 20, 26这几种MMP之外的其他MMP家族成员在两种细胞中均有表达,而MMP-7仅在内侧副韧带成纤维细胞中被检测出有表达,TIMPs家族的4个成员在两种细胞中均有表达。实时定量荧光PCR检测结果显示在前交叉韧带成纤维细胞中MMP-1, 2, 14, 17, 23A, 23B及TIMP-4的表达量明显高于在内侧副韧带成纤维细胞中的表达。由此,我们得出结论:MMP家族与TIMPs家族中大部分成员在两种细胞中均有表达,且存在差异,这个现象有可能是二者愈合能力不同的原因之一。更多还原

【Abstract】 The human ACL (anterior cruciate ligament) is susceptible to injury but has poor healing response, whereas an injured MCL (medial collateral ligament) can be repaired relatively well. Since MMPs (matrix metalloproteases) and TIMPs (tissue inhibitor of metalloproteases) are involved in this tissue remodeling process, investigation of different response of MMPs and TIMPs family in ACL and MCL fibroblasts might lead to understanding the differential matrix remodeling process as well as their different healing ability. The first step would be determination of whether these tissue remodeling effectors are present in ligaments. In this study, we designed primers for real-time RT-PCR and determined the expression of MMPs and TIMPs family in ACL and MCL fibroblasts with synovium as a positive control. Semiquantitative RT-PCR revealed that multiple MMPs and TIMPs expressed in human ACL and MCL fibroblasts except MMP-8, 10, 12, 13, 15, 16, 20, and 26. MMP-7 was present in MCL but not in ACL fibroblast.Quantitative real-time RT-PCR showed that mRNA levels of MMP-1, 2, 14, 17, 23A, and 23B and TIMP-4 are significantly higher in MCL than in ACL fibroblasts. However, MMP-3 is higher in ACL than in MCL fibroblasts. We conclude that numerous MMPs and TIMPs family members that are differentially expressed in ACL and MCL might be involved in the differential matrix remodeling process as well as the differential healing ability of ACL and MCL.更多还原