Human Neutrophil Defensins and Their Effect on Epithelial Cells
Human Neutrophil Defensins and Their Effect on Epithelial Cells
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DOI：
10.1902/jop.2012.120017
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发表时间：
2013-01-01
影响因子：
4.3
通讯作者：
Uitto, Veli-Jukka
中科院分区：
文献类型：
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作者：
Gursoy, Ulvi K.;Kononen, Eija;Uitto, Veli-Jukka
背景：本研究的目的包括：1)通过HNP-3在牙龈和中性粒细胞提取物处理的上皮细胞单层中定位人中性粒细胞防御素-1 (HNP-1)；2)确定HNP-1对上皮细胞活力、附着和扩散的影响；3)分析HNP-1对细菌粘附上皮细胞的影响。方法：采用免疫组织化学和免疫细胞化学方法定位龈组织和预培养单层细胞中HNP-1和HNP-3的位置。不同HNP-1浓度（低，1 ~ 5 μ g/mL，中，10 μ g/mL，高，20 ~ 50 μ g/mL）培养角质形成细胞后，用商用试剂盒检测上皮细胞的活力和增殖。显微镜下观察在HNP-1存在下，角化细胞在纤维连接蛋白包被表面的附着和扩散情况。用标准抗生素试验测定核梭菌ATCC25586和普雷沃菌中间ATCC25611在HNP-1预孵育的角质形成细胞上的附着。结果：HNP-1至HNP-3在临床健康牙龈的交界处上皮和牙周炎牙龈的袋上皮中均有分布。当角质形成细胞单层与中性粒细胞提取物孵育时，HNP-1到HNP-3被结合到生长的细胞集落的周围。在低HNP-1浓度下，角质形成细胞增殖增强。中、高浓度的HNP-1显著增加细胞死亡。HNP-1以浓度依赖的方式增加了角化细胞在纤维连接蛋白包被表面上的附着和扩散，以及细菌对角化细胞的附着。结论：HNP-1通过刺激角质形成细胞的增殖、附着和扩散，对角质形成细胞的完整性起作用，而高剂量会增加细菌附着和角质形成细胞的死亡。中华牙周病杂志（英文版）；2013;31(4):391 - 391。
Background: The aims of the present study include: 1) to localize human neutrophil defensin-1 (HNP-1) through HNP-3 in gingiva and in neutrophil extract-treated epithelial cell monolayers; 2) to determine the effects of HNP-1 on the epithelial cell viability, attachment, and spreading; and 3) to analyze the effect of HNP-1 on the bacterial adherence to epithelial cells.Methods: For localization of HNP-1 through HNP-3 in gingival tissue and in preincubated cell monolayers, immunohistochemical and immunocytochemical methods were used. Viability and proliferation of epithelial cells were determined with commercial kits after incubating the keratinocytes with different HNP-1 concentrations (low, 1 to 5 mu g/mL; moderate, 10 mu g/mL; high, 20 to 50 mu g/mL). Attachment and spreading of keratinocytes on fibronectin-coated surfaces in the presence of HNP-1 were evaluated under microscope. Attachment of Fusobacterium nucleatum ATCC25586 and Prevotella intermedia ATCC25611 on keratinocytes preincubated with HNP-1 were determined with a standard antibiotic test.Results: HNP-1 through HNP-3 localized in the junctional epithelium of clinically healthy gingiva and in the pocket epithelium of gingiva with periodontitis. When keratinocyte monolayers were incubated with neutrophil extracts, HNP-1 through HNP-3 were bound to the periphery of the growing cell colonies. In low HNP-1 concentrations, the keratinocyte proliferation enhanced. Moderate and high concentrations of HNP-1 increased the cellular death significantly. HNP-1 increased the attachment and spreading of keratinocytes on fibronectin-coated surfaces and bacterial attachment to keratinocytes in a concentration-dependent manner.Conclusion: HNP-1 plays a role in the integrity of keratinocytes by stimulating their proliferation, attachment, and spreading, whereas higher doses increase the bacterial attachment and keratinocyte death. J Periodontol 2013;84:126-133.