华西口腔医学杂志2025，Vol.43Issue(6)：808-818,11.DOI:10.7518/hxkq.2025.2024391
龈沟液中细胞外游离DNA与牙周临床指标及环磷酸鸟苷—磷酸腺苷合成酶—干扰素基因刺激因子信号通路相关分子的关联性分析
Correlation analysis of cell-free DNA in gingival crevicular fluid with periodontal clinical indicators and cyclic guanosine phosphate-adenosine phosphate synthase-stimulator of interferon genes signaling pathway
摘要
Abstract
Objective This study aims to explore the potential relationships of cell-free DNA(cfDNA)in gingival cre-vicular fluid(GCF)with periodontal clinical indicators and the expression of DNA receptor pathway cyclic guanosine phosphate-adenosine phosphate synthase(cGAS)-stimulator of interferon genes(STING)in gingival tissues and human gingival fibroblasts(HGFs).Methods GCF and gingival tissue samples were collected from periodontally healthy indi-viduals and patients diagnosed with periodontitis.Periodontal clinical indicators were recorded,including plaque index(PLT),bleeding index(BI),probing depth(PD),and clinical attachment level(CAL).The concentration of cfDNA in GCF was quantified,and the correlation between GCF and periodontal clinical indicators was analyzed.Immunofluorescence and reverse transcription-quantitative polymerase chain reaction(RT-qPCR)were used to assess the distribution of cGAS,STING,and p-STING in gingival tissues.Additionally,the mRNA expression levels of the key components of the cGAS-STING signaling pathway,namely,cGAS,STING,inhibitory of kappa-B kinase(IKK),nuclear factor kappa-B p65(NF-κB p65),interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α),were measured.Furthermore,cfDNA extracted from GCF was employed to stimulate HGFs in the healthy control and periodontitis groups,and the mRNA expression lev-els of the key molecules of cGAS-STING signaling pathway were detected through Western blot and RT-qPCR.Results The concentration of cfDNA in GCF was found to be significantly elevated in the periodontitis group compared with the control group.Moreover,cfDNA concentration demonstrated a strong positive correlation with the periodontal clinical indi-cators.Immunofluorescence analysis revealed considerably increased percentage of fluorescence co-localization of cGAS,STING,and p-STING with the gingival fibroblast FSP-1 marker in the gingival tissues of the periodontitis group.The mRNA expression levels of cGAS,STING,IKK,NF-κB p65,IL-1β,IL-6,and TNF-α were significantly higher in the peri-odontitis group.In vitro stimulation of HGFs with GCF-derived cfDNA resulted in increased protein expression of cGAS and p-STING and considerably upregulated the mRNA expression levels of cGAS,STING,IKK,NF-κB p65,IL-1β,IL-6,and TNF-α in the healthy and periodontitis groups compared with the blank group.Correlation analysis showed that the concentration of cfDNA at the sampling site was positively correlated with the mRNA expression levels of cGAS,STING,NF-κB p65,and IL-6 in gingival tissues.Conclusion cfDNA concentrations in the GCF of patients with periodontitis are considerably elevated,and are associated with the activation of the cGAS-STING signaling pathway in HGFs.These find-ings suggest that cfDNA contributes to the progression of periodontitis.关键词
牙周炎/细胞外游离DNA/环磷酸鸟苷—磷酸腺苷合成酶—干扰素基因刺激因子信号通路/人牙龈成纤维细胞Key words
periodontitis/cell-free DNA/cyclic guanosine phosphate-adenosine phosphate synthase-stimulator of interferon genes signaling pathway/human gingival fibroblasts分类
医药卫生引用本文复制引用
陈兰,朱轩智,周婕妤,李继遥,赵蕾..龈沟液中细胞外游离DNA与牙周临床指标及环磷酸鸟苷—磷酸腺苷合成酶—干扰素基因刺激因子信号通路相关分子的关联性分析[J].华西口腔医学杂志,2025,43(6):808-818,11.基金项目
国家自然科学基金(81970944,81991501,81991502) （81970944,81991501,81991502）
四川省自然科学基金(2023NSFSC0553) National Natural Science Foundation of China(81970944,81991501,81991502) （2023NSFSC0553）
Natural Science Foun-dation of Sichuan Province(2023NSFSC0553) （2023NSFSC0553）
