Hou W S, Li Z, Gordon R E, Chan K, Klein M J, Levy R, Keysser M, Keyszer G, Brömme D
Department of Human Genetics, Mount Sinai School of Medicine, New York, New York 10029-6574, USA.
Am J Pathol. 2001 Dec;159(6):2167-77. doi: 10.1016/S0002-9440(10)63068-4.
Synovial fibroblasts (SFs) play a critical role in the pathogenesis of rheumatoid arthritis (RA) and are directly involved in joint destruction. Both SF-resident matrix metalloproteases and cathepsins have been implicated in cartilage degradation although their identities and individual contributions remain unclear. The aims of this study were to investigate the expression of cathepsin K in SFs, the correlation between cathepsin K expression and disease severity, and the contribution of cathepsin K to fibroblast-mediated collagen degradation. Immunostaining of joint specimens of 21 patients revealed high expression of cathepsin K in SFs in the synovial lining and the stroma of synovial villi, and to a lesser extent in CD68-positive cells of the synovial lining. Cathepsin K-positive SFs were consistently observed at sites of cartilage and bone degradation. Expression levels of cathepsin K in the sublining and vascularized areas of inflamed synovia showed a highly significant negative correlation with results derived from the Hannover Functional Capacity Questionnaire (r = 0.78, P = 0.003; and r = 0.70, P = 0.012, respectively) as a measure of the severity of RA in individual patients. For comparison, there was no correlation between Hannover Functional Capacity Questionnaire and cathepsin S whose expression is limited to CD-68-positive macrophage-like synoviocytes. The expression of cathepsin K was also demonstrated in primary cell cultures of RA-SFs. Co-cultures of SFs on cartilage disks revealed the ability of fibroblast-like cells to phagocytose collagen fibrils whose intralysosomal hydrolysis was prevented in the presence of a potent cathepsin K inhibitor but not by an inhibitor effective against cathepsins L, B, and S. The selective and critical role of cathepsin K in articular cartilage and subchondral bone erosion was further corroborated by the finding that cathepsin K has a potent aggrecan-degrading activity and that cathepsin K-generated aggrecan cleavage products specifically potentiate the collagenolytic activity of cathepsin K toward type I and II collagens. This study demonstrates for the first time a critical role of cathepsin K in cartilage degradation by SFs in RA that is comparable to its well-known activity in osteoclasts.
滑膜成纤维细胞(SFs)在类风湿性关节炎(RA)的发病机制中起关键作用,并直接参与关节破坏。尽管其具体身份和各自作用尚不清楚,但SFs中的基质金属蛋白酶和组织蛋白酶均与软骨降解有关。本研究的目的是调查组织蛋白酶K在SFs中的表达、组织蛋白酶K表达与疾病严重程度之间的相关性,以及组织蛋白酶K对成纤维细胞介导的胶原蛋白降解的作用。对21例患者的关节标本进行免疫染色显示,滑膜衬里和滑膜绒毛基质中的SFs中组织蛋白酶K表达较高,滑膜衬里的CD68阳性细胞中表达较低。在软骨和骨降解部位始终观察到组织蛋白酶K阳性的SFs。炎症滑膜的衬里下层和血管化区域中组织蛋白酶K的表达水平与汉诺威功能能力问卷的结果呈高度显著负相关(分别为r = 0.78,P = 0.003;r = 0.70,P = 0.012),该问卷用于衡量个体患者RA的严重程度。作为对照,汉诺威功能能力问卷与组织蛋白酶S之间无相关性,组织蛋白酶S的表达仅限于CD-68阳性的巨噬细胞样滑膜细胞。在RA-SFs的原代细胞培养中也证实了组织蛋白酶K的表达。在软骨盘上进行SFs的共培养显示,成纤维样细胞具有吞噬胶原纤维的能力,在存在强效组织蛋白酶K抑制剂的情况下,其溶酶体内水解受到抑制,但对组织蛋白酶L、B和S有效的抑制剂则无此作用。组织蛋白酶K对关节软骨和软骨下骨侵蚀具有选择性和关键作用,这一发现进一步得到证实,即组织蛋白酶K具有强大的聚集蛋白聚糖降解活性,且组织蛋白酶K产生的聚集蛋白聚糖裂解产物可特异性增强组织蛋白酶K对I型和II型胶原的胶原olytic活性。本研究首次证明了组织蛋白酶K在RA中SFs介导的软骨降解中起关键作用,这与其在破骨细胞中众所周知的活性相当。