二极管激光对牙龈卟啉单胞菌脂多糖刺激下人牙龈成纤维细胞增殖和炎症反应的影响

Effect of diode laser on the proliferation and inflammation of Porphyromonas gingivalis LPS-treated human gingival fibroblast cells

目的:探讨二极管激光对牙龈卟啉单胞菌(Porphyromonas gingivalis,P.gingivalis)脂多糖(lipopolysaccharide,LPS)刺激下的人牙龈成纤维细胞(human gingival fibroblast cells,HGFs)增殖和炎症反应的影响,初步揭示二极管激光对慢性牙周炎的作用机制。方法:原代培养HGFs;不同浓度的LPS和2μg/mL LPS+二极管激光作用于HGFs 24、48、72 h后,细胞增殖实验检测增殖能力变化;2μg/mL LPS,2μg/mL LPS+二极管激光作用于HGFs 24 h后,免疫荧光实验观察波形蛋白(Vimentin)的表达,实时定量PCR检测炎症因子白介素-6(interleukin-6,IL-6)和IL-8的表达。结果:运用组织块贴壁法培养出HGFs;与对照组相比,LPS抑制HGFs的增殖,但二极管激光改善LPS对HGFs增殖影响,促进HGFs的增殖;LPS能够提高HGFs中IL-6和IL-8的表达,抑制Vimentin的表达,二极管激光可显著逆转LPS刺激下HGFs中IL-6、IL-8和Vimentin的表达水平。结论:二极管激光可逆转P.gingivalis LPS对HGFs的增殖作用及其对炎症反应影响,二极管激光能够抑制LPS在慢性牙周炎过程中的作用。

Objective:To investigate the role of diode laser on the proliferation and inflammation of Porphyromonas gingivalis lipopolysaccharide(LPS)-treated human gingival fibroblast cells(HGFs),and discovered the mechanisms of diode laser in the process of periodontitis.Methods:Primary HGFs were isolated from human periodontal tissues.Treatment with LPS under different concentrations or 2μg/mL LPS combined with diode laser for 24,48 and 72 h,the proliferation assay were performed to investigate the proliferation activity of HGFs.After treatment with LPS or 2μg/mL LPS combined with diode laser,immunofluorescence assay was used to detect the expression of Vimentin in HGFs,and real-time PCR was performed to analyze the expression of IL-6 and IL-8 in HGFs.Results:HGFs were identified using immunocytochemical methods.Incubation with LPS decreased the proliferation of HGFs.Compared with LPS,treatment with LPS and diode laser significantly enhanced the proliferation of HGFs,and treatment with LPS and diode laser decreased the expression of IL-6,IL-8,and increased Vimentin in HGFs.Conclusion:Diode laser can enhance the proliferation of LPS-treated HGFs,and decreased the inflammatory activity of LPS-treated HGFs.Studies pave the way for the clinical use of diode laser in periodontitis.