Aggrecan degradation in human cartilage - Evidence for both matrix metalloproteinase and aggrecanase activity in normal, osteoarthritic, and rheumatoid joints
Aggrecan degradation in human cartilage - Evidence for both matrix metalloproteinase and aggrecanase activity in normal, osteoarthritic, and rheumatoid joints
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人体软骨中的凝集素降解--正常关节、骨关节炎关节和类风湿关节中基质金属蛋白酶和凝集素酶活性的证据
DOI：
10.1172/jci119526
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发表时间：
1997-07-01
影响因子：
15.9
通讯作者：
Lohmander, LS
中科院分区：
文献类型：
--
作者：
Lark, MW;Bayne, EK;Lohmander, LS
为了检测基质金属蛋白酶（MMPs）和聚集蛋白聚糖酶在对照和患病的人关节软骨中的活性，用单特异性抗肽抗体检测聚集蛋白聚糖的代谢片段。比较MMP产生的终止于VDIPEN 341的聚集蛋白聚糖G1片段的分布和数量与聚集蛋白聚糖酶产生的终止于NITEGE（373）的G1片段的分布。大小与球间结构域区域中的单一酶促切割一致，这些片段没有进一步的蛋白水解加工。两个新表位也通过免疫组织化学在来自骨关节炎（OA）、类风湿性关节炎（RA）的接受关节置换的患者的关节软骨和来自没有已知关节疾病的成人的软骨中检测到。在对照样本中，两个G1片段的染色强度随着年龄增加，在来自22周龄胎儿样本的软骨中几乎没有染色。MMP产生的新表位的提取量相对于聚集蛋白聚糖和胶原蛋白含量也随着年龄的增加而增加，证实了免疫组织化学结果。在20-30岁之后，这种关系保持在稳定状态。MMP产生的表位的染色在显示损伤的组织学迹象的对照软骨中最显著，而聚集蛋白聚糖酶产生的片段的强染色经常在缺乏明显组织学损伤的成人软骨中被注意到。两种新表位的强烈染色出现在组织的更严重的原纤维化的浅表区域中。在来自OA或RA患者的关节软骨中也检测到VDIPEN-和NITEGE-新表位的强烈免疫染色，这些标本中的软骨素显著更降解，并且在具有广泛软骨损伤的区域中总是观察到两种表位的高水平染色。OA和RA样品中提取的VDIPEN新表位相对于胶原蛋白或聚集蛋白聚糖的水平与年龄匹配的对照标本中观察到的相似。在细胞周围观察到两种类型的聚集蛋白聚糖片段的免疫染色，但在区域间基质中也进一步去除。在组织的一些区域中，发现了两个新表位，而在其他区域中仅检测到一个。因此，这些特定分解代谢聚集蛋白聚糖片段的产生和/或周转不一定是协调的。我们的结果与基于经典MMP和聚集蛋白聚糖酶的针对聚集蛋白聚糖的蛋白水解活性在正常和关节炎关节软骨中的存在一致。''
To examine the activity of matrix metalloproteinases (MMPs) and aggrecanase in control and diseased human articular cartilage, metabolic fragments of aggrecan were detected with monospecific antipeptide antibodies. The distribution and quantity of MMP-generated aggrecan G1 fragments terminating in VDIPEN341 were compared with the distribution of aggrecanase-generated G1 fragments terminating in NITEGE(373), Both types of G1 fragments were isolated from osteoarthritic cartilage. The sizes were consistent with a single enzymatic cleavage in the interglobular domain region, with no further proteolytic processing of these fragments. Both neoepitopes were also detected by immunohistochemistry in articular cartilage from patients undergoing joint replacement for osteoarthritis (OA), rheumatoid arthritis (RA), and in cartilage from adults with no known joint disease,In control specimens, the staining intensity for both G1 fragments increased with age, with little staining in cartilage from 22-wk-old fetal samples. There was also an increase with age in the extracted amount of MMP-generated neoepitope in relation to both aggrecan and collagen content, confirming the immunohistochemical results, After the age of 20-30 yr this relationship remained at a steady state, The staining for the MMP-generated epitope was most marked in control cartilage exhibiting histological signs of damage, whereas intense staining for the aggrecanase-generated fragment was often noted in adult cartilage lacking overt histological damage. Intense staining for both neoepitopes appeared in the more severely fibrillated, superficial region of the tissue,Intense immunostaining for both VDIPEN- and NITEGE-neoepitopes was also detected in joint cartilage from patients with OA or RA, Cartilage in these specimens was significantly more degraded and high levels of staining for both epitopes was always seen in areas with extensive cartilage damage. The levels of extracted VDIPEN neoepitope relative to collagen or aggrecan in both OA and RA samples were similar to those seen in age-matched control specimens,Immunostaining for both types of aggrecan fragments was seen surrounding the cells but also further removed in the interterritorial matrix. In some regions of the tissue, both neoepitopes were found while in others only one was detected, Thus, generation and/or turnover of these specific catabolic aggrecan fragments is not necessarily coordinated, Our results are consistent with the presence in both normal and arthritic joint cartilage of proteolytic activity against aggrecan based on both classical MMPs and ''aggrecanase.''