Weckroth M, Vaheri A, Lauharanta J, Sorsa T, Konttinen Y T
Department of Virology, Haartman Institute, University of Helsinki, Finland.
J Invest Dermatol. 1996 May;106(5):1119-24. doi: 10.1111/1523-1747.ep12340167.
Although extracellular proteolysis is a prerequisite for normal wound healing, uncontrolled proteolytic tissue destruction appears to be a pathogenic factor in non-healing wounds. The aim of our study was to compare the activities of the serine proteinases of polymorphonuclear origin, elastase and cathepsin G, and the metalloproteinases, gelatinase and collagenase, in chronic leg ulcer exudate (10 patients) and acute wound fluid (6 patients). Serine proteinase activities were low in leg ulcer exudates but very high in some but not all acute wound fluids. Total collagenase activity, measured as activity against type I collagen monitored by SDS-PAGE and densitometry, was higher in chronic leg ulcer exudate than in acute wound fluid and its degree of autoactivation was relatively high. Doxycycline inhibition studies suggested that the collagenase activity in chronic leg ulcer exudate was MMP-1 ("fibroblast-type") and not MMP-8 ("neutrophil-type"). Zymographic analysis of the gelatinolytic enzymes in acute wound fluid showed a progressive increase from the day of operation to postoperative day 5, but the degree of activity was lower than in chronic leg ulcer exudate and the low molecular mass activation products were faint. The leg ulcer gelatinase profiles were characterized by high expression of 92/82- and 72/62-kDa duplex bands and by the presence of low molecular mass activation products. Leg ulcer collagenase seems to be derived from mononuclear rather than polymorphonuclear cells, which are known to be involved in acute wound healing. In conclusion, the present study shows that gelatinase and collagenase, but not elastase and cathepsin G are found in chronic leg ulcer exudate.
尽管细胞外蛋白水解是正常伤口愈合的先决条件，但不受控制的蛋白水解组织破坏似乎是难愈合伤口的致病因素。我们研究的目的是比较多形核来源的丝氨酸蛋白酶（弹性蛋白酶和组织蛋白酶G）以及金属蛋白酶（明胶酶和胶原酶）在慢性腿部溃疡渗出液（10例患者）和急性伤口液（6例患者）中的活性。腿部溃疡渗出液中的丝氨酸蛋白酶活性较低，但在一些而非所有急性伤口液中活性非常高。通过SDS-PAGE和光密度法监测，以针对I型胶原的活性来衡量的总胶原酶活性在慢性腿部溃疡渗出液中高于急性伤口液，并且其自身激活程度相对较高。强力霉素抑制研究表明，慢性腿部溃疡渗出液中的胶原酶活性是MMP-1（“成纤维细胞型”）而非MMP-8（“中性粒细胞型”）。对急性伤口液中明胶分解酶的酶谱分析显示，从手术当天到术后第5天活性逐渐增加，但活性程度低于慢性腿部溃疡渗出液，且低分子量激活产物较淡。腿部溃疡明胶酶谱的特征是92/82-kDa和72/62-kDa双链带的高表达以及低分子量激活产物的存在。腿部溃疡胶原酶似乎来源于单核细胞而非已知参与急性伤口愈合的多形核细胞。总之，本研究表明慢性腿部溃疡渗出液中存在明胶酶和胶原酶，但不存在弹性蛋白酶和组织蛋白酶G。
