Adhesion of monocytes to periodontal fibroblasts requires activation of NOD1/2-and TLR4-mediated LFA-1 and VLA-4
Adhesion of monocytes to periodontal fibroblasts requires activation of NOD1/2-and TLR4-mediated LFA-1 and VLA-4
复制标题
单核细胞与牙周成纤维细胞的粘附需要激活 NOD1/2 和 TLR4 介导的 LFA-1 和 VLA-4。
DOI：
10.1016/j.archoralbio.2014.12.012
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发表时间：
2015-06-01
影响因子：
3
通讯作者：
Ouyang, Xiangying
中科院分区：
文献类型：
--
作者：
Liu, Jianru;Liu, Wenyi;Ouyang, Xiangying
目的：本研究的目的是探讨含核苷酸结合寡聚化结构域蛋白1/2（NOD 1/2）和Toll样受体4（TLR 4）在介导单核细胞通过白细胞功能相关抗原1（LFA-1）和极晚期抗原4（VLA-4）与牙周成纤维细胞粘附中的作用。采用免疫组化法检测慢性牙周炎患者牙龈组织中NOD 1、NOD 2和TLR 4的表达。分别用NOD 1、NOD 2和TLR 4激动剂处理U937细胞24 h，或用针对NOD 1、NOD 2和TLR 4的小干扰RNA（siRNA）转染U937细胞48 h，观察U937细胞与人牙龈成纤维细胞（hGFs）和人牙周膜细胞（hPDLCs）的粘附情况。同时通过实时荧光定量PCR、Western blot和流式细胞术检测U937细胞中LFA-1和VLA-4的表达。结果：NOD 1、NOD 2和TLR 4在牙周炎患者牙龈组织中均呈高表达，尤其是在单核细胞浸润区。通过激动剂激活这些受体，上调U937细胞中LFA-1和VLA-4的表达，并增加U937细胞与hGFs或hPDLCs的亲和力。另一方面，通过特异性siRNA敲低这些受体导致相反的结果。此外，阻断U937细胞中的LFA-1或VLA-4可显著减弱激动剂触发的U937与牙周成纤维细胞的粘附（P
Objective: The aim of this study was to investigate the roles of nucleotide-binding oligomerization domain-containing protein 1/2 (NOD1/2) and Toll-like receptor 4 (TLR4) in mediating the adhesion of monocytes to periodontal fibroblasts through leucocyte function-associated antigen-1 (LFA-1) and very late antigen-4 (VLA-4).Design: The expression of NOD1, NOD2, and TLR4 was detected in the gingival tissue of patients with chronic periodontitis by immunohistochemistry. Then the adhesion of cells of human monocytic cell line U937 to human gingival fibroblasts (hGFs) and human periodontal ligament cells (hPDLCs) was investigated after U937 cells were treated with the agonists of NOD1, NOD2, and TLR4 for 24h, or transfected with small interfering RNAs (siRNAs) targeting NOD1, NOD2, and TLR4 for 48h. Meanwhile, the expression of LFA-1 and VLA-4 was examined in U937 cells through real-time polymerase chain reaction (PCR), Western blot, and flow cytometry. To confirm the roles of LFA-1 and VLA-4 involved in the process of adhesion, the adhesion blockade assay was performed using the corresponding blocking antibodies against these adhesion molecules.Results: The immunostaining results showed that NOD1, NOD2, and TLR4 were highly expressed in the gingival tissue of patients with periodontitis, especially in the monocyte-infiltrated area. The activation of these receptors by agonists upregulated the expression of LFA-1 and VLA-4 in U937 cells, and it increased the affinity of U937 cells to hGFs or hPDLCs. On the other hand, knockdown of these receptors by specific siRNAs resulted in the opposite results. In addition, blocking either LFA-1 or VLA-4 in U937 cells significantly attenuated the agonist-triggered adhesion of U937 to periodontal fibroblasts (P