CD14-mediated signal pathway of Porphyromonas gingivalis lipopolysaccharide in human gingival fibroblasts
CD14-mediated signal pathway of Porphyromonas gingivalis lipopolysaccharide in human gingival fibroblasts
复制标题
人牙龈成纤维细胞中CD14介导的牙龈卟啉单胞菌脂多糖信号通路
作者：
A. Watanabe;Akira Takeshita;S. Kitano;S. Hanazawa
脂多糖（LPS）通过CD 14诱导单核细胞/巨噬细胞中炎性细胞因子的表达，CD 14是LPS受体之一，其主要在这些细胞中表达。牙龈卟啉单胞菌脂多糖（P-LPS）也能诱导人牙龈成纤维细胞产生炎性细胞因子。因此，确定牙龈成纤维细胞中是否表达CD 14并确定细胞中P-LPS介导的信号转导机制是重要的。在这项研究中，我们观察到出乎意料的免疫组织化学，蛋白质印迹（免疫印迹），和北方（RNA）印迹分析，CD 14在人牙龈成纤维细胞高密度表达。P-LPS诱导的单核细胞趋化蛋白1（MCP-1）基因在细胞中的表达被抗人CD 14抗体处理显着抑制，并完全抑制除草霉素A，一种有效的酪氨酸激酶抑制剂。该抑制剂还显著抑制单核细胞趋化活性和细胞产生MCP-1。此外，细胞中P-LPS诱导的MCP-1基因表达也被两种转录因子的抑制剂阻断，即，AP-1抑制剂姜黄素和NF-κ B抑制剂吡咯烷二硫代氨基甲酸酯。这两种抑制剂抑制单核细胞趋化活性的培养上清液中的P-LPS处理的细胞。凝胶迁移率分析显示P-LPS处理的细胞中AP-1和NF-κ B含量增加。本研究首次证实了CD 14在人牙龈成纤维细胞中的表达，并表明P-LPS在细胞中的信号转导途径是由CD 14介导的。
Lipopolysaccharide (LPS) induces expression of inflammatory cytokines in monocytes/macrophages via CD14, one of the LPS receptors, which is expressed predominantly in these cells. It has been demonstrated that Porphyromonas gingivalis LPS (P-LPS) also is able to induce inflammatory cytokines in human gingival fibroblasts. Therefore, it is important to determine whether CD14 is expressed in gingival fibroblasts and to define the P-LPS-mediated signal-transducing mechanism in the cells. In this study, we observed unexpectedly by immunohistochemical, Western blotting (immunoblotting), and Northern (RNA) blotting assays that CD14 is expressed at high density in human gingival fibroblasts. P-LPS-induced expression of the monocyte chemoattractant protein 1 (MCP-1) gene in the cells was inhibited markedly by treatment with anti-human CD14 antibody and was completely inhibited by herbimycin A, a potent inhibitor of tyrosine kinase. The inhibitor also dramatically inhibited monocyte chemotactic activity of and MCP-1 production by the cells. Furthermore, P-LPS-induced expression of the MCP-1 gene in the cells also was blocked by inhibitors of two transcription factors, i.e., curcumin, an inhibitor of AP-1, and pyrolidine dithiocarbamate, an inhibitor of NF-kappaB. Both inhibitors inhibited monocyte chemotactic activity in the culture supernatant of P-LPS-treated cells. Gel shift mobility assay showed stimulation of the AP-1 and NF-kappaB contents in P-LPS-treated cells. This study is the first to demonstrate the expression of CD14 in human gingival fibroblasts and to show that the signal-transducing pathway of P-LPS in the cells is mediated by CD14.
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影响因子：
56.9
作者：
WRIGHT, SD;RAMOS, RA;MATHISON, JC
通讯作者：
MATHISON, JC
蛋白激酶 C 和蛋白酪氨酸激酶参与脂多糖诱导的人单核细胞产生 TNF-α 和 IL-1β。
DOI：
--
发表时间：
1994
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Shapira,L;Takashiba,S;Champagne,C;Amar,S;VanDyke,TE
通讯作者：
VanDyke,TE
DOI：
10.1016/s0021-9258(19)74564-5
发表时间：
1993-11
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Jiahuai Han;Jiing-Dwan Lee;P. Tobias;R. Ulevitch
通讯作者：
Jiahuai Han;Jiing-Dwan Lee;P. Tobias;R. Ulevitch
由脂多糖诱导的 CD14 介导的核因子-κ B 易位不需要酪氨酸激酶活性。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Delude,RL;Fenton,MJ;SavedraJr,R;Perera,PY;Vogel,SN;Thieringer,R;Golenbock,DT
通讯作者：
Golenbock,DT
影响因子：
4.4
作者：
A. Haziot;B. Tsuberi;S. Goyert
通讯作者：
A. Haziot;B. Tsuberi;S. Goyert