Chang Lien-Yu, Lai Yu-Lin, Yu Tzu-Hsuan, Chen Yen-Ting, Hung Shan-Ling
Institute of Oral Biology, National Yang-Ming University, Taipei, Taiwan.
J Periodontol. 2014 Jun;85(6):859-67. doi: 10.1902/jop.2013.130198. Epub 2013 Sep 3.
Areca chewers have a higher prevalence of periodontitis than non-chewers. Cell adhesion and movement (migration) are important for leukocyte recruitment to inflammation sites. This study investigates the effects of areca nut extract (ANE) on the adhesion and migration abilities of the human immune cells, peripheral blood mononuclear cells (PBMCs). The combined effects of nicotine and lipopolysaccharides (LPS) were also analyzed.
Purified PBMCs obtained from healthy adults were treated with ANE, nicotine, and/or LPS. Cell adhesion ability was examined using fibronectin-coated microslides, Liu stain, and light microscopy. Cell migration ability was evaluated using the transwell system followed by staining and fluorescence microscopy. Statistical difference was analyzed using the Mann-Whitney U test.
When compared with the media-treated control samples, PBMCs treated with ANE for 4 hours showed a significant reduction of the adherent cells on the microslides. Interestingly, LPS treatment increased cell adhesion, which could be reduced by simultaneous ANE plus nicotine treatment. The chemotactic migration of PBMCs was reduced by ANE treatment for 1, 4, or 24 hours in a dose-dependent manner. LPS treatment increased PBMC migration, which could be reduced by simultaneous treatment with ANE or with ANE plus nicotine.
ANE reduced the adhesion and migration abilities of PBMC. ANEs, with or without nicotine, also attenuated the migration of LPS-stimulated PBMCs. The results implicated that the immune cell functions were impaired in areca chewers, which might increase the host susceptibility to oral and periodontal infection.
嚼槟榔者患牙周炎的几率高于不嚼槟榔者。细胞黏附与移动(迁移)对于白细胞募集至炎症部位十分重要。本研究调查槟榔提取物(ANE)对人体免疫细胞即外周血单核细胞(PBMCs)黏附与迁移能力的影响。同时也分析了尼古丁与脂多糖(LPS)的联合作用。
从健康成年人中获取纯化的PBMCs,用ANE、尼古丁和/或LPS进行处理。使用纤连蛋白包被的载玻片、刘氏染色法和光学显微镜检查细胞黏附能力。使用Transwell系统,随后进行染色和荧光显微镜检查来评估细胞迁移能力。采用曼-惠特尼U检验分析统计学差异。
与培养基处理的对照样本相比,用ANE处理4小时的PBMCs在载玻片上的贴壁细胞显著减少。有趣的是,LPS处理增加了细胞黏附,而ANE加尼古丁同时处理可使其降低。ANE处理1、4或24小时以剂量依赖方式降低了PBMCs的趋化迁移。LPS处理增加了PBMC迁移,而ANE或ANE加尼古丁同时处理可使其降低。
ANE降低了PBMC的黏附与迁移能力。无论有无尼古丁,ANE均减弱了LPS刺激的PBMCs的迁移。结果表明嚼槟榔者的免疫细胞功能受损,这可能会增加宿主对口腔和牙周感染的易感性。