Gelatinase A (MMP-2) activation by skin fibroblasts: dependence on MT1-MMP expression and fibrillar collagen form.
Gelatinase A (MMP-2) activation by skin fibroblasts: dependence on MT1-MMP expression and fibrillar collagen form.
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皮肤成纤维细胞激活明胶酶 A (MMP-2)：依赖于 MT1-MMP 表达和纤维状胶原蛋白形式。
DOI：
10.1016/s0945-053x(01)00135-4
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Thompson,EW
中科院分区：
文献类型：
--
作者：
Ruangpanit,N;Chan,D;Holmbeck,K;Birkedal-Hansen,H;Polarek,J;Yang,C;Bateman,JF;Thompson,EW
进一步探讨了原代成纤维细胞培养物激活 MMP-2 时胶原蛋白和 MT1-MMP 各自的需求。富含人类 I 型和 III 型胶原蛋白或由重组人 II 型或 III 型胶原蛋白组成的三维凝胶可导致 MT1-MMP 产生（mRNA 和蛋白质）增加并诱导 MMP-2 激活。使用干燥的单体胶原仅观察到边缘诱导，证实需要胶原纤维组织来活化。令我们惊讶的是，添加到成纤维细胞培养物中的相对少量（低至 25 μg/ml）的酸溶性 I 型胶原蛋白也诱导了有效的 MMP-2 激活。然而，通过将胶原蛋白与原纤维阻断肽预温育时所观察到的抑制以及已知会损害原纤维化的碱处理胶原制剂所观察到的活化减少来表明添加的胶原蛋白对胶原原纤维形成的需要。用高碘酸钠预处理胶原蛋白也消除了 MMP-2 激活诱导。当在培养物中添加不形成胶原纤维的IV型胶原时，缺乏活化提供了胶原纤维形成需要的进一步证据。与同窝对照小鼠相比，MT1-MMP 缺陷小鼠的成纤维细胞无法响应三维胶原凝胶或添加的胶原溶液而激活 MMP-2。总的来说，这些数据表明胶原蛋白和 MT1-MMP 的纤维结构对于成纤维细胞中 MMP-2 激活反应至关重要。
The respective requirements of collagen and MT1-MMP in the activation of MMP-2 by primary fibroblast cultures were explored further. Three-dimensional gels enriched in human collagen types I and III or composed of recombinant human type II or III collagen, caused increased MT1-MMP production (mRNA and protein) and induced MMP-2 activation. Only marginal induction was seen with dried monomeric collagen confirming the need for collagen fibrillar organisation for activation. To our surprise, relatively low amounts (as low as 25 μg/ml) of acid soluble type I collagen added to fibroblast cultures also induced potent MMP-2 activation. However, the requirement for collagen fibril formation by the added collagen was indicated by the inhibition seen when the collagen was pre-incubated with a fibril-blocking peptide, and the reduced activation seen with alkali-treated collagen preparations known to have impaired fibrilisation. Pre-treatment of the collagen with sodium periodate also abrogated MMP-2 activation induction. Further evidence of the requirement for collagen fibril formation was provided by the lack of activation when type IV collagen, which does not form collagen fibrils, was added in the cultures. Fibroblasts derived from MT1-MMP-deficient mice were unable to activate MMP-2 in response to either three-dimensional collagen gel or added collagen solutions, compared to their littermate controls. Collectively, these data indicate that the fibrillar structure of collagen and MT1-MMP are essential for the MMP-2 activational response in fibroblasts.