Involvement of adhesion molecule in in vitro plaque-like formation of macrophages stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide
Involvement of adhesion molecule in in vitro plaque-like formation of macrophages stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide
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DOI：
10.1111/j.1600-0765.2010.01270.x
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发表时间：
2010-08-01
影响因子：
3.5
通讯作者：
Tatsuji, N.
中科院分区：
文献类型：
--
作者：
Takeshi, T.;Keisuke, N.;Tatsuji, N.
背景与目的：牙周袋内的炎症因子如脂多糖（LPS）可通过激活白细胞促进动脉粥样硬化的形成。在我们之前的研究中，我们开发了一种微通道芯片来观察流体系统中的细胞粘附过程。本次调查的目的是研究牙周病细菌脂多糖增强斑块样形成的微通道chipe.Material和方法的机制：要评估的粘附分子，如细胞间粘附分子1（ICAM-1），淋巴细胞功能相关抗原1（LFA-1）和L-选择素，在小鼠巨噬细胞RAW 264.7细胞的表面上的表达，放线菌伴聚杆菌脂多糖的影响，每个粘附分子的表达进行了检查，通过流式细胞术和蛋白质印迹分析。结果：LPS作用12 h后，RAW264.7细胞表面ICAM-1和LFA-1表达增加，L-selectin表达无明显变化。免疫印迹分析也证实了ICAM-1表达的增加。流动试验显示抗ICAM-1抗体可抑制LPS刺激的巨噬细胞在微通道芯片微柱上形成噬斑样。结论：ICAM-1在LPS刺激的巨噬细胞形成噬斑样中起重要作用。我们的微通道芯片是一个合适的工具，动脉粥样硬化，包括牙周炎，在体外的病因调查。
Background and Objective:Inflammatory agents, such as lipopolysaccharide (LPS), in periodontal pockets may promote atherogenesis by activating leukocytes. In our previous study, we developed a microchannel chip to observe the cell adhesion process in a fluid system. The objective of this investigation was to examine the mechanism by which periodontopathic bacterial LPS enhances plaque-like formation on a microchannel chip.Material and Methods:To evaluate the effect of Aggregatibacter actinomycetemcomitans LPS on the expression of adhesion molecules, e.g. intercellular adhesion molecule 1 (ICAM-1), lymphocyte function-associated antigen 1 (LFA-1) and L-selectin, on the surface of murine macrophage RAW264.7 cells, the expression of each adhesion molecule was examined by flow cytometry and western blot analysis. Moreover, a flow test on the microchannel chip involving anti-adhesion molecule antibodies was conducted to clarify which adhesion molecule is related to plaque-like formation of RAW264.7 cells.Results:The expressions of ICAM-1 and LFA-1 on the surface of RAW 264.7 cells increased following 12 h culture with LPS; L-selectin expression was unaffected. An increase in ICAM-1 expression was also confirmed by western blot analysis. The flow test revealed that anti-ICAM-1 antibody inhibited plaque-like formation of LPS-stimulated macrophages on the micropillars of the microchannel chip.Conclusion:These findings indicate that ICAM-1 plays an important role in plaque-like formation of LPS-stimulated macrophages. Our microchannel chip is a suitable tool for the investigation of etiological factors of atherosclerosis, including periodontitis, in vitro.