alpha(1)-proteinase inhibitor in gingival crevicular fluid of humans with adult periodontitis: Serpinolytic inhibition by doxycycline
alpha(1)-proteinase inhibitor in gingival crevicular fluid of humans with adult periodontitis: Serpinolytic inhibition by doxycycline
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DOI：
10.1111/j.1600-0765.1997.tb01377.x
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发表时间：
1997-01-01
影响因子：
3.5
通讯作者：
Crout, R
中科院分区：
文献类型：
--
作者：
Lee, HM;Golub, LM;Crout, R
血清蛋白，α(1)-蛋白酶抑制剂（α (1)-PI），保护宿主免受丝氨酸蛋白酶，如PMN弹性酶的侵害。利用兔抗人α (1)-PI血清，采用点印迹法构建标准曲线，定量测定GCF中的α (1)-PI蛋白。分别从健康(H)、牙龈炎(G)和成人牙周炎（AP）患者的滤纸上收集GCF，然后用Tris/NaCl/CaCl2缓冲液（pH 7.6）提取。α (1)-PI浓度随G升高而升高，AP组最高。H位点仅显示完整的α (1)-PI (52 kDa)；未检测到48 kDa的降解片段。在G和AP受试者中，分别在17%和71%的GCF样本中发现α (1)-PI降解片段。胶原酶和α (1)- pi降解活性随着炎症的严重程度而增加（GCF流量）。此外，α (1)-PI降解（或丝氨酸溶解）活性被表征为基质金属蛋白酶，可能是胶原酶，基于其对一系列不同蛋白酶抑制剂（包括强力霉素）的体外反应。我们建议:(1)胶原酶不仅通过降解胶原蛋白促进牙周破坏，还通过消耗调节弹性蛋白酶和其他丝氨酸蛋白酶的α (1)-PI，从而有利于更广泛地攻击细胞外基质（ECM）成分；(2)基于最近一项纵向双盲研究，使用上述α (1)-PI分析技术，成人牙周炎患者服用低剂量强力霉素可以抑制ECM降解的广泛级联反应。
The serum protein, alpha(1)-proteinase inhibitor (alpha(1)-PI), defends the host against serine proteinases, e.g. PMN elastase. Using a rabbit anti-serum against human alpha(1)-PI, this protein in GCF was quantified from a standard curve constructed from dot-blot analysis and characterized by Western blot. GCF was collected on filter paper strips from healthy (H), gingivitis (G) and adult periodontitis (AP) patients, then extracted with Tris/NaCl/CaCl2 buffer, pH 7.6. alpha(1)-PI concentration increased with G and was highest in AP subjects. H sites only showed intact alpha(1)-PI (52 kDa); no degradation fragments (48 kDa) were detected. In G and AP subjects, alpha(1)-PI degradation fragments were seen in 17% and 71% of GCF samples, respectively. Both collagenase and alpha(1)-PI-degrading activities in GCF increased with severity of inflammation (GCF flow). Moreover, the alpha(1)-PI degrading (or serpinolytic) activity was characterized as a matrix metalloproteinase, probably collagenase, based on its in vitro response to a panel of different proteinase inhibitors including doxycycline. We propose: (1) that collagenase promotes periodontal breakdown not only by degrading collagen, but also by depleting alpha(1)-PI regulation of elastase and other serine-proteinases, thereby favoring a broader attack on extracellular matrix (ECM) constituents, and (2) based on a recent longitudinal double-blind study using the techniques described above for alpha(1)-PI analysis, that low-dose doxycycline administration to humans with adult periodontitis can inhibit this broad cascade of ECM degradation.