内毒素刺激人牙龈成纤维细胞表达mCD14和TLR4的免疫细胞化学研究

Expression of mCD14 and TLR4 in human gingival fibroblast stimulated with E.coli lipopolysaccharide

目的:检测内毒素刺激后体外培养人牙龈成纤维细胞mCD14和TLR4的表达,探讨牙龈成纤维细胞作为免疫活性细胞在牙周炎发生发展中的作用。方法:组织块法原代培养人牙龈成纤维细胞并进行来源鉴定。以0.1、1、5、10μg/mL内毒素刺激第4代细胞,培养24h后用免疫细胞化学染色法检测mCD14和TLR4在牙龈成纤维细胞中的表达,用多功能彩色细胞分析管理系统进行图像分析并进行统计学处理。结果:正常人牙龈成纤维细胞mCD14和TLR4表达均为弱阳性,经内毒素刺激后,mCD14和TLR4均呈强阳性表达,且随内毒素浓度的增加而染色增强,各实验组染色强度均显著高于对照组(P<0.01)。结论:内毒素刺激可使牙龈成纤维细胞表达mCD14和TLR4增强,提示牙龈成纤维细胞具有免疫细胞的功能特性并参与局部牙周组织炎症。

AIM： To examine the expression of mCD14 and Toll-like vecehtor 4 （TLR4） in human gingival fibroblasts （HGFs） stimulated by E coli lipopolysaccharide （LPS） and to provide new insight about the role of human gingival fibroblasts as immunocompetent cells in the development of periodontitis. METHODS： Human gingival fibroblasts were cultured and immunologically identified. The fourth passage cells were inoculated onto glass covers and stimu- lated with LPS at different concentration for 24h. Then immunocytochemistry was calxied out to examine the expression of mCD14 and TLR4 in the cells. RESULTS： Immunocytochemistry revealed that mCD14 and TLR4 were expressed only mildly in normal human gingival fibroblasts. After 24h stimulation with LPS, both mCD14 and TLR4 were strongly ex- pressed, and the staining intensity was increased with the LPS concentration （P 〈 0.01 ）. CONCLUSION： Human gin- gival fibroblasts express mCD14 and TLR4 under LPS stimulation, indicating that it has the characteristics of inununocom- petent cells.