Inhibitory effects of tumor necrosis factor-alpha on migration of human periodontal ligament cells.
Inhibitory effects of tumor necrosis factor-alpha on migration of human periodontal ligament cells.
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DOI：
10.1902/jop.2006.050192
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发表时间：
2006-05
影响因子：
4.3
通讯作者：
A. Takemura;I. Nakagawa;S. Kawai;H. Inaba;Takahiro Kato;S. Hamada;A. Amano
中科院分区：
文献类型：
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作者：
A. Takemura;I. Nakagawa;S. Kawai;H. Inaba;Takahiro Kato;S. Hamada;A. Amano
肿瘤坏死因子-α（TNF-α）与慢性牙龈炎症有关，并被怀疑影响牙周破坏。然而，TNF-α在伤口愈合和牙周组织再生中的确切作用在很大程度上是未知的。在本研究中，我们研究了TNF-α对人牙周膜细胞（PDL）迁移和增殖的影响。方法体外培养人牙周膜细胞，观察TNF-α对细胞迁移和增殖的影响。研究了α 5和β 1整合素亚基及其相关分子桩蛋白和粘着斑激酶（FAK）的蛋白表达谱。同时检测纤维连接蛋白基因表达。此外，使用GTP负载下拉测定法评估Rho家族小鸟苷三磷酸（GTP）结合蛋白（RhoA）的活化，并且还使用共聚焦显微镜观察转染paxillin融合的绿色荧光蛋白（GFP）的表达载体后PDL细胞的粘着斑形成。结果TNF-α可损伤细胞迁移，加入抗TNF-α抗体后细胞迁移恢复。相反，PDL细胞增殖不受TNF-α的影响。TNF-α上调α 5和β 1整合素亚单位的表达，而纤连蛋白没有过表达。PDL细胞可诱导桩蛋白和FAK磷酸化，并诱导RhoA活化。共聚焦显微镜分析显示，TNF-α诱导粘着斑和应力纤维形成的所有部分的细胞。结论TNF-α通过增强细胞粘附能力，抑制细胞迁移，促进粘着斑形成和应力纤维形成。
BACKGROUND Tumor necrosis factor-alpha (TNF-alpha) is associated with chronic gingival inflammation and is suspected to influence periodontal destruction. However, the exact roles of TNF-alpha in wound healing and periodontal tissue regeneration are largely unknown. In the present study, we examined the effects of TNF-alpha on migration and proliferation of human periodontal ligament (PDL) cells. METHODS PDL cells were cultured in the presence of TNF-alpha to determine its effects on cellular migration and proliferation. The protein expression profiles of alpha5 and beta1 integrin subunits and their related molecules, paxillin and focal adhesion kinases (FAK), were investigated. Gene expression of fibronectin also was assayed. Further, the activation of Rho-family small guanosine triphosphate (GTP)-binding protein (RhoA) was evaluated using a GTP-loading pull-down assay, and focal adhesion formation by PDL cells after transfection with the expression vector of paxillin-fused green fluorescent protein (GFP) also was observed with confocal microscopy. RESULTS Cellular migration was impaired by TNF-alpha and recovered following the addition of anti-TNF-alpha antibodies. In contrast, PDL cell proliferation was not affected by TNF-alpha. TNF-alpha upregulated the expression of the alpha5 and beta1 integrin subunits, whereas fibronectin was not overexpressed. Phosphorylation of paxillin and FAK by PDL cells was induced, and RhoA activation also was induced. Confocal microscopic analysis revealed that TNF-alpha induced focal adhesion and stress fiber formation in all parts of the cells. CONCLUSION Our results suggested that TNF-alpha impairs cellular migration by enhancing cellular adhesive ability following significant focal adhesion and stress fiber formation.