COLLAGENASE AND COLLAGENASE INHIBITORS IN OSTEOARTHRITIC AND NORMAL HUMAN CARTILAGE
COLLAGENASE AND COLLAGENASE INHIBITORS IN OSTEOARTHRITIC AND NORMAL HUMAN CARTILAGE
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DOI：
10.1172/jci108632
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发表时间：
1977-01-01
影响因子：
15.9
通讯作者：
VIGLIANI, G
中科院分区：
文献类型：
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作者：
EHRLICH, MG;MANKIN, HJ;VIGLIANI, G
在晚期骨关节炎中，关节表面的所有软骨成分都会丢失。虽然蛋白多糖的降解机制是存在的，但目前还没有已知的去除胶原蛋白的系统。胶原成分的丢失可能是关节软骨胶原酶的功能之一。正常人软骨中的这种酶与一种抑制物结合在一起，似乎只有很少量的存在。试图证明胶原酶活性在磨碎的人关节软骨或其溶酶体部分没有成功。7天的软骨组织培养也未能证明该酶的存在；同样的培养液与胰酶孵育后显示胶原显著降解，表明胰酶破坏了该抑制物。7天的培养液在肝素充满的Sepharose4B亲和层析柱上进行层析，该亲和层析用溴化氰活化。这去除了抑制物，骨关节炎软骨的层析液释放了42%的胶原底物结合计数，但正常软骨释放了10.1%，而胰酶对照组释放了6.4%。37℃孵育的酶-胶原复合体降解产物的电泳法。C显示破裂完全为可透析的小碎片，而在25度。C较大的碎片被剥离。
In advanced osteoarthritis, all of the cartilaginous components are lost from the joint surface. Although mechanisms exist for proteoglycan degradation, no system is known for removal of the collagen. The loss of the collagen components may be a function of articular cartilage collagenase. The enzyme in normal human cartilage is bound to an inhibitor and appears to be present in very small amounts. Attempts to demonstrate collagenase activity in ground human articular cartilage or in its lysosomal fraction were unsuccessful. Seven-day cartilage tissue cultures also failed to demonstrate the presence of the enzyme; the same culture fluid, incubated with trypsin, showed significant degradation of collagen, suggesting that trypsin destroyed the inhibitor. Seven-day culture fluids were then chromatographed on a heparin-charged Sepharose 4B affinity column that was activated with cyanogen bromide. This removed the inhibitor, and the chromatographed fluid from osteoarthritic cartilage released 42% of the incorporated counts of the collagen substrate, but normal cartilage released 10.1% and a trypsin control, 6.4%. Electrophoresis of the degradation products of the enzyme-collagen complex incubated at 37.degree. C revealed breakdown was complete to small dialyzable fragments, while at 25.degree. C larger fragments were split off.