Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin
Impaired degradation of matrix collagen in human gingival fibroblasts by the antiepileptic drug phenytoin
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DOI：
10.1902/jop.2005.76.6.941
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发表时间：
2005-06-01
影响因子：
4.3
通讯作者：
Amano, A
中科院分区：
文献类型：
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作者：
Kato, T;Okahashi, N;Amano, A
背景：牙龈过度生长（GO）是与苯妥英（PHT）给药相关的严重不良反应，PHT诱导的牙龈过度生长的特点是牙龈结缔组织中细胞外基质成分，特别是胶原蛋白的大量积累。然而，这种胶原蛋白积累的病因仍然很大程度上未知。我们研究了 PHT 对导致人牙龈成纤维细胞 (HGF) 中胶原蛋白积累的胶原蛋白降解过程的影响。方法：用不同浓度的 PHT 培养 HGF，并测定活细胞数和胶原蛋白量。分别通过逆转录聚合酶链式反应 (RT-PCR) 分析和蛋白质印迹法对基质金属蛋白酶 (MMP) 和 MMP 组织抑制剂 (TIMP) 的基因和蛋白质表达进行定量。使用流式细胞术分析测定胶原蛋白的细胞内吞作用。检测PHT对细胞外信号调节激酶1/2（ERK1/2）和抑制剂κB-α（IκB-α）的影响。结果：PHT不影响HGF的增殖，但显着增加胶原蛋白的积累。此外，MMP-1、-2和-3的表达被PHT显着抑制，而TIMP-1的表达以剂量和时间依赖性方式被诱导。 PHT 还显着阻止 HGF 的胶原蛋白内吞作用，这与抑制 α2β1-整合素表达有关。此外，PHT 抑制了 ERK1/2 的磷酸化和 I kappa B-α 降解。结论：这些结果表明，PHT 通过抑制 MMP/TIMP-1 和 α2β1-整合素介导的内吞作用的酶促降解，导致胶原蛋白降解受损。这些改变可能是通过 ERK1/2 和核因子 kappa B 的细胞信号通路介导的。这些协同作用可能会导致胶原蛋白积累，从而导致 GO。
Background: Gingival overgrowth (GO) is a serious adverse effect associated with the administration of phenytoin (PHT), with PHT-induced GO characterized by a massive accumulation of extracellular matrix components, especially collagen, in gingival connective tissues. However, the etiology of such collagen accumulation is still largely unknown. We examined the effects of PHT on the Collagen degradation process leading to collagen accumulation in human gingival fibroblasts (HGF).Methods: HGFs were cultured with various concentrations of PHT and viable cell numbers and collagen amounts were determined. Gene and protein expressions of matrix metalloproteinases (MMP) and tissue inhibitors of MMPs (TIMP) were quantified with reverse transcription-polymerase chain reaction (RT-PCR) analyses and Western blotting, respectively. Cellular endocytosis of collagen was assayed using flow-cytometric analysis. The effects of PHT on extracellular signal-regulated kinase 1/2 (ERK1/2) and inhibitor kappa B-alpha (I kappa B-alpha) were assayed.Results: The proliferation of HGFs was not affected by PHT, whereas it significantly increased collagen accumulation. Further, the expressions of MMP- 1, -2, and -3 were markedly suppressed by PHT, whereas that of TIMP-1 was induced in a dose- and time-dependent manner. PHT also markedly prevented Collagen endocytosis by HGFs, which was associated with the suppression of alpha 2 beta 1-integrin expression. In addition, the phosphorylation of ERK1/2 and I kappa B-alpha degradation were suppressed by PHT.Conclusions: These results suggest that PHT causes an impaired degradation of collagen by suppression of enzymatic degradation with MMPs/TIMP-1 and alpha 2 beta 1-integrin-mediated endocytosis. Those alterations are likely mediated through the cellular signaling pathways of ERK1/2 and nuclear factor kappa B. These synergistic effects may cause collagen accumulation, leading to GO.