Activation of neutrophil collagenase in periodontitis
Activation of neutrophil collagenase in periodontitis
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DOI：
10.1128/iai.67.5.2319-2326.1999
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发表时间：
1999-05-01
影响因子：
3.1
通讯作者：
McCulloch, CAG
中科院分区：
文献类型：
--
作者：
Romanelli, R;Mancini, S;McCulloch, CAG
中性粒细胞胶原酶（基质金属蛋白酶 8 [MMP-8]）是炎症性疾病中组织破坏的重要介质。厌氧性牙周感染的研究表明，龈沟液中的活性 MhP-8 与进行性牙周炎中牙周组织的降解有关，而潜伏酶在牙龈炎中占主导地位。由于 MMP-8 的激活似乎是牙周炎的关键步骤，因此我们使用可溶性生物素化胶原基质检测了龈沟液样品中 MMP-8 的激活。对牙周炎、牙龈炎和对照的龈沟液的分析显示，牙周炎 (n = 12) 样本中的活性胶原酶水平比牙龈炎 (n = 17) 样本高六倍 (P < 0.001)，牙龈炎 (n = 17) 样本的活性水平较低，而对照 (n = 25) 则没有活性。 收集龈沟液后，没有进一步激活潜在的胶原酶 尽管MMP-1和MMP-8都可以通过免疫印迹检测到，但不能检测到MMP-13，但MMP-1的封闭抗体表明胶原酶活性很大程度上是由MMP-8贡献的，MMP-8位于病变组织的基质中。龈沟液中的 MMP-8 主要以 60 kDa 形式迁移，并有少量 78 kDa 种类，而从外周中性粒细胞中分离出的 MMP-8 以 70 kDa 和 89 kDa 迁移，分别对应于酶的活性形式和潜在形式。 60 kDa 和 70 kDa 条带中的大多数 MMP-8 选择性结合金属蛋白酶 2 和胶原蛋白的组织抑制剂，表明这些条带中的大多数（但不是全部）酶处于激活形式。然而，不同样本中龈沟液中 78 kDa 和 60 kDa 形式的量与胶原酶测定测量的潜在酶和活性酶不相关 (r(2) = 0.028)。总的来说，这些研究已经确定了龈沟液中不同形式的潜在和活性 MMP-8，它们似乎是牙周炎中发生的独特激活机制的结果。 MMP-8 激活的复杂性通过分别从龈沟液和中性粒细胞样本中获得的 60 kDa 和 70 kDa 条带中存在的 MMP-8 潜在、激活和超激活形式进一步表明。
Neutrophil collagenase (matrix metalloproteinase 8 [MMP-8]) is an important mediator of tissue destruction in inflammatory diseases. Studies of anaerobic periodontal infections have shown that active MhP-8 in gingival crevicular fluid is associated with the degradation of periodontal tissues in progressive periodontitis whereas the latent enzyme is predominant in gingivitis. Since the activation of MMP-8 appears to be a crucial step in periodontitis, we have examined the activation of MMP-8 in gingival crevicular fluid samples by using a soluble biotinylated collagen substrate. Analysis of gingival crevicular fluid in periodontitis, gingivitis, and controls revealed sixfold (P < 0.001)-higher levels of active collagenase in periodontitis (n = 12) samples compared to gingivitis (n = 17) samples, which exhibited low levels of activity, while controls (n = 25) showed no activity, After gingival crevicular fluid was collected, no further activation of latent collagenase occurred in vitro, Although both MMP-1 and MMP-8, but not MMP-13, could be detected by immunoblots, blocking antibodies to MMP-1 showed that collagenase activity was largely contributed by MMP-8, which was localized to the matrix of diseased tissues. The MMP-8 in gingival crevicular fluid migrated primarily as a 60-kDa form with smaller amounts of a 78-kDa species, whereas MMP-8 isolated from peripheral neutrophils migrated at 70 and 89 kDa, corresponding to active and latent forms of the enzyme, respectively. Most of the MMP-8 in the 60- and 70-kDa bands selectively bound to tissue inhibitor of metalloproteinase 2 and collagen, indicating that most, but not all, of the enzyme in these bands was in an activated form. However, the amounts of the 78- and 60-kDa forms from gingival crevicular fluid in different samples did not correlate (r(2) = 0.028) with the latent and active enzyme measured by collagenase assay. Collectively, these studies have identified distinct forms of latent and active MMP-8 in gingival crevicular fluid that appear to result from a unique activation mechanism that occurs in periodontitis. The complexity of MMP-8 activation is further indicated by the presence of latent, activated, and superactivated forms of MMP-8 in the 60- and 70-kDa bands obtained from gingival crevicular fluid and neutrophil samples, respectively.