Effect of Galardin on collagen degradation by Pseudomonas aeruginosa
Effect of Galardin on collagen degradation by Pseudomonas aeruginosa
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DOI：
10.1006/exer.1999.0755
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发表时间：
1999-12-01
影响因子：
3.4
通讯作者：
Nishida, T
中科院分区：
文献类型：
--
作者：
Hao, JL;Nagano, T;Nishida, T
作者研究了一种人工合成的基质金属蛋白酶抑制剂Galardin在角质形成细胞存在或不存在的情况下对铜绿假单胞菌(P.uginosa)降解胶原的影响。将含有或不含有悬浮角质形成细胞的I型胶原凝胶在含有无菌铜绿假单胞菌培养液和/或Galardin的培养液中孵育24小时。培养过程中胶原纤维的降解通过羟脯氨酸的释放来衡量。用明胶酶谱和Western blotting分析铜绿假单胞菌因子(S)对角质形成细胞释放的基质金属蛋白酶(MMPs)的激活作用。研究了抑肽酶、亮肽酶和胃抑素对铜绿假单胞菌降解胶原蛋白的影响。铜绿假单胞菌发酵液本身可引起胶原胶降解。当角质形成细胞存在时，铜绿假单胞菌肉汤进一步增加了胶原的降解量。Galardin显著减少了铜绿假单胞菌发酵液降解的胶原量，无论凝胶中是否存在角质形成细胞。然而，这些蛋白水解酶抑制剂对胶原降解没有抑制作用。明胶酶谱和Western blotting显示，在铜绿假单胞菌发酵液的存在下，角质形成细胞释放的失活的前基质金属蛋白酶-1、-2和-3被转化为活性形式。Galardin可降低活性MMPs的数量，增加失活的proMMPs的数量，提示Galardin可抑制铜绿假单胞菌对proMMPs的激活。本研究结果提示Galardin通过抑制原MMPs向活性MMPs的转化，从而抑制绿脓杆菌培养液中角质形成细胞介导的胶原降解。(C)1999年学术出版社。
The authors examined the effect of a synthetic peptidyl hydroxamate inhibitor of matrix metalloproteinase, Galardin, on collagen degradation by Pseudomonas aeruginosa (P. aeruginosa) in the presence or absence of keratocytes. Type I collagen gels, with or without suspended keratocytes, were incubated under medium containing sterile P. aeruginosa culture broth and/or Galardin for 24 hr. Degradation of collagen fibrils during culture was measured by the release of hydroxyproline. The conditioned media were also subjected to gelatin zymography and Western blotting to analyse the activation, by P. aeruginosa factor(s), of matrix metalloproteinases (MMPs) released by keratocytes. The effects of protease inhibitors, aprotinin, leupeptin and pepstatin, on collagen degradation by P. aeruginosa were also examined. P. aeruginosa broth by itself induced collegen gel degradation. When keratocytes were present, P. aeruginosa broth increased the amount of degraded collagen even further. Galardin significantly reduced the amounts of collagen degraded by P. aeruginosa culture broth, whether keratocytes were present or absent in the gel. However, the protease inhibitors had no inhibitory effects on collagen degradation. Gelatin zymography and Western blotting revealed that inactive proMMP-1, -2 and -3, released by keratocytes, were converted to active forms in the presence of P. aeruginosa broth. Galardin decreased the amounts of active MMPs and increased those of inactive proMMPs, suggesting that Galardin inhibited the activation of proMMPs by P. aeruginosa. The present results suggest that Galardin inhibits the keratocyte-mediated collagen degradation by P. aerginosa culture broth, resulting from preventing the conversion of proMMPs to active MMPs. (C) 1999 Academic Press.