百里香酚通过调控炎症通路抑制脂多糖诱导的大鼠牙龈成纤维细胞炎症的研究

doi:10.13701/j.cnki.kqyxyj.2025.09.008

口腔医学研究 ›› 2025, Vol. 41 ›› Issue (9): 781-787.DOI: 10.13701/j.cnki.kqyxyj.2025.09.008

百里香酚通过调控炎症通路抑制脂多糖诱导的大鼠牙龈成纤维细胞炎症的研究

郭嘉¹, 高梦洁¹, 牛慧¹, 刘丹枫², 陈曦^1*

1.郑州大学第一附属医院郑东院区口腔内科郑州河南 450052;
2.郑州大学第一附属医院牙体牙髓科郑州河南 450052

收稿日期:2025-02-13 出版日期:2025-09-28 发布日期:2025-09-24
通讯作者: * 陈曦,E-mail:2423074549@qq.com
作者简介:郭嘉(1983~ ),男,河南焦作人,博士,副主任医师,研究方向:牙周病学及口腔遗传病学。
基金资助:
河南省医学科技攻关计划联合共建项目基金(2023年度)(编号:LHGJ20230242)

Thymol Inhibits LPS-induced Inflammation in Rat Gingival Fibroblasts by Regulating Inflammatory Pathways

GUO Jia¹, GAO Mengjie¹, NIU Hui¹, LIU Danfeng², CHEN Xi^1*

1. Department of Stomatology, Zhengdong Branch of the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China;
2. Department of Endodontics and Endodontics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China

Received:2025-02-13 Online:2025-09-28 Published:2025-09-24

摘要/Abstract

摘要： 目的: 探讨百里香酚(Thymol)对牙龈卟啉单胞菌(Porphyromonas gingivalis,P. g)的影响,和对脂多糖(lipopolysaccharide,LPS)诱导的大鼠牙龈成纤维细胞炎症反应的抑制作用。方法: 采用微量稀释法测定Thymol对P. g的最低抑菌浓度(minimum inhibitory concentration,MIC)和杀菌浓度(minimum bactericidal concentration,MBC);生物膜粘附减少-结晶紫法测定Thymol对P. g的抗生物膜活性;扫描电子显微镜和共聚焦激光扫描显微镜(confocal scanning laser microscopy,CSLM)检测细菌。通过大鼠牙龈成纤维细胞建立5组牙周炎体外实验模型:对照组、LPS+不同浓度Thymol(0、10、20、40 μg/mL)组。通过ELISA测定上清中相关蛋白的表达,包括肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)、骨保护素(osteoclastogenesis inhibitory factor,OPG)和核因子kappa-B配体的可溶性受体激活因子(soluble receptor activator of nuclear factor-κB ligand,sRANKL),并计算OPG/sRANKL比值;采用Western blot检测牙龈成纤维细胞核蛋白中NF-κB p65和IκBα的磷酸化水平。结果: Thymol减弱了P. g的毒性和生物膜形成。与LPS组相比,Thymol刺激后,细胞上清液中炎症因子TNF-α、IL-1β和IL-6的表达水平显著降低,而抗炎因子IL-10的表达水平显著升高;OPG/sRANKL比值增加;LPS诱导的NF-κB p65和IκBα的磷酸化水平显著降低(P＜0.001)。结论: Thymol通过抑制P. g的毒性和生物膜形成,调控相关炎症因子的表达(TNF-α、IL-1β、IL-6、IL-10、OPG/sRANKL)及炎症通路(NF-κB p65、IκBα),抑制LPS诱导的大鼠牙龈成纤维细胞的炎症反应。

关键词: 百里香酚, 牙龈卟啉单胞菌, 脂多糖, 牙龈成纤维细胞, NF-κB 通路

Abstract: Objective: To investigate the effect of thymol on Porphyromonas gingivalis (P. g) and lipopolysaccharide (LPS)-induced inflammatory response in rat gingival fibroblasts. Methods: The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of thymol against P. g were determined by microdilution method. Biofilm adhesion reduction-crystal violet assay was used to determine the anti-biofilm activity of thymol against P. g. Bacteria were detected by scanning electron microscopy and confocal scanning laser microscopy (CSLM). Rat gingival fibroblasts were used to establish periodontitis models in vitro: control group, LPS + different concentrations of thymol (0, 10, 20, and 40 μg/mL) groups. ELISA was used to measure the expression of related proteins in the supernatant, including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), osteoclastogenesis inhibitory factor (OPG) and soluble receptor activator of nuclear factor-κB ligand (sRANKL), and OPG/sRANKL ratio was calculated. Western blot was used to detect the phosphorylation of NF-κB p65 and IκBα in the nuclear protein of gingival fibroblasts. Results: Thymol attenuated P. g toxicity and biofilm formation. Thymol had no adverse effect on LPS-induced growth of rat gingival fibroblasts. Compared with LPS group, the expression levels of inflammatory factors TNF-α, IL-1β, and IL-6 in the cell supernatant were significantly decreased after Thymol stimulation, while the expression level of anti-inflammatory factor IL-10 was significantly increased. The ratio of OPG/sRANKL was increased. LPS-induced phosphorylation of NF-κB p65 and IκBα was significantly decreased(P＜0.001). Conclusion: Thymol inhibits P. g toxicity and biofilm formation, and inhibits LPS-induced inflammation in GFS by regulating the expression of related inflammatory factors (TNF-α, IL-1β, IL-6, IL-10, OPG/sRANKL) and inflammatory pathways (NF-κB p65, IκBα).

Key words: thymol, Porphyromonas gingivalis, lipopolysaccharide, gingival fibroblasts, NF-κB signaling pathway

郭嘉, 高梦洁, 牛慧, 刘丹枫, 陈曦. 百里香酚通过调控炎症通路抑制脂多糖诱导的大鼠牙龈成纤维细胞炎症的研究[J]. 口腔医学研究, 2025, 41(9): 781-787.

GUO Jia, GAO Mengjie, NIU Hui, LIU Danfeng, CHEN Xi. Thymol Inhibits LPS-induced Inflammation in Rat Gingival Fibroblasts by Regulating Inflammatory Pathways[J]. Journal of Oral Science Research, 2025, 41(9): 781-787.

参考文献

[1] Bui FQ, Almeida-da-Silva CLC, Huynh B, et al. Association between periodontal pathogens and systemic disease[J]. Biomed J, 2019, 42(1):27-35.
[2] Dye BA. Global periodontal disease epidemiology[J]. Periodontol 2000, 2012, 58(1):10-25.
[3] Li YY, Cai Q, Li BS, et al. The effect of Porphyromonas gingivalis lipopolysaccharide on the pyroptosis of gingival fibroblasts[J]. Inflammation, 2021, 44(3):846-858.
[4] Sahoo CR, Paidesetty SK, Padhy RN. The recent development of thymol derivative as a promising pharmacological scaffold[J]. Drug Dev Res, 2021, 82(8):1079-1095.
[5] Marchese A, Orhan IE, Daglia M, et al. Antibacterial and antifungal activities of thymol: A brief review of the literature[J]. Food Chem, 2016, 210:402-414.
[6] Sapkota M, Li L, Kim SW, et al. Thymol inhibits RANKL-induced osteoclastogenesis in RAW264.7 and BMM cells and LPS-induced bone loss in mice[J]. Food Chem Toxicol, 2018, 120:418-429.
[7] Alfonso García SL, Parada-Sanchez MT, Arboleda Toro D. The phenotype of gingival fibroblasts and their potential use in advanced therapies[J]. Eur J Cell Biol, 2020, 99(7):151123.
[8] Jentsch HF, Eckert FR, Eschrich K, et al. Antibacterial action of chlorhexidine/thymol containing varnishes in vitro and in vivo[J]. Int J Dent Hyg, 2014, 12(3):168-173.
[9] Chatrath A, Gangwar R, Kumari P, et al. In vitro anti-biofilm activities of citral and thymol against Candida tropicalis[J]. J Fungi (Basel), 2019, 5(1):13.
[10] Kirkwood KL, Li F, Rogers JE, et al. A p38alpha selective mitogen-activated protein kinase inhibitor prevents periodontal bone loss[J]. J Pharmacol Exp Ther, 2007, 320(1):56-63.
[11] Naruishi K, Nagata T. Biological effects of interleukin-6 on gingival fibroblasts: Cytokine regulation in periodontitis[J]. J Cell Physiol, 2018, 233(9):6393-6400.
[12] Lopez-Castejon G, Brough D. Understanding the mechanism of IL-1β secretion[J]. Cytokine Growth Factor Rev, 2011, 22(4):189-195.
[13] Pan W, Wang Q, Chen Q. The cytokine network involved in the host immune response to periodontitis[J]. Int J Oral Sci, 2019, 11(3):30.
[14] Saraiva M, O'Garra A. The regulation of IL-10 production by immune cells[J]. Nat Rev Immunol, 2010, 10(3):170-181.
[15] Sojod B, Chateau D, Mueller CG, et al. RANK/RANKL/OPG signalization implication in periodontitis: New evidence from a RANK transgenic mouse model[J]. Front Physiol, 2017, 8:338.
[16] Liu B, Wu Y, Wang Y, et al. NF-κB p65 knock-down inhibits TF, PAI-1 and promotes activated protein C production in lipopolysaccharide-stimulated alveolar epithelial cells type Ⅱ[J]. Exp Lung Res, 2018, 44(4-5):241-251.

百里香酚通过调控炎症通路抑制脂多糖诱导的大鼠牙龈成纤维细胞炎症的研究

Thymol Inhibits LPS-induced Inflammation in Rat Gingival Fibroblasts by Regulating Inflammatory Pathways

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	顾菀蓉, 冉蓓, 赵奕雯, 廖金凤. g-C₃N₄基复合材料在牙周炎中的光催化/光热抗菌治疗研究[J]. 口腔医学研究, 2025, 41(9): 773-780.
[2]	熊曼雯, 刘敏. 牙龈素在心血管疾病中致病机制的研究进展[J]. 口腔医学研究, 2025, 41(7): 547-551.
[3]	郭雅彤, 张正, 陈溯, 张研. 3D打印钛合金表面氢化TiO₂纳米管对人牙龈成纤维细胞生物学行为影响的实验研究[J]. 口腔医学研究, 2025, 41(7): 607-614.
[4]	张彩霞, 周怡文, 闻娟, 黄子维, 林爽, 杨任, 李煌, 李贵凤. 卵泡激素相互作用蛋白1对牙周炎症的调控研究[J]. 口腔医学研究, 2025, 41(6): 521-528.
[5]	魏巍, 谭小容, 李慕秋, 龚忠诚, 李晨曦. 牙龈卟啉单胞菌通过NDK/ATP/P2X7影响口腔鳞状细胞癌恶性表型的实验研究[J]. 口腔医学研究, 2025, 41(5): 378-385.
[6]	张黄, 苗瑞婧, 范旭升, 黄婕, 王永武. 探究NLRP3炎症小体对人原代牙龈成纤维细胞炎症刺激的影响[J]. 口腔医学研究, 2025, 41(2): 128-133.
[7]	向景, 陈祁航, 强晓叶, 王舒欣, 张得钧. 基于网络药理学和体外实验验证研究复方金银花防治牙周炎的作用机制[J]. 口腔医学研究, 2025, 41(1): 26-34.
[8]	唐路, 杨文文, 张献丽, 张欣然, 薛栋, 冷丽君, 赵颖. TRAF6/Runx2信号轴对牙龈卟啉单胞菌诱导血管平滑肌细胞钙化调控作用的研究[J]. 口腔医学研究, 2024, 40(5): 401-406.
[9]	庞晓婵, 陈青立, 雒国凤, 尚政军. 牙龈卟啉单胞菌对口腔鳞状细胞癌化疗耐药行为的影响[J]. 口腔医学研究, 2024, 40(5): 413-416.
[10]	李俊俊, 王雯, 郭慧颖, 袁长永, 夏廷旭, 王鹏来. 3D生物打印构建HGFs和HUVECs共培养体系促进HUVECs成血管[J]. 口腔医学研究, 2024, 40(4): 297-303.
[11]	姜震, 颜燕宏, 韩雪, 蒋备战. 大肠杆菌脂多糖对大鼠牙囊干细胞生物学行为的影响[J]. 口腔医学研究, 2023, 39(6): 534-540.
[12]	张福, 周平, 向锋, 汪春艳, 李镭. 硒代蛋氨酸通过NF-κB和MAPK通路抑制P.g-LPS诱导的RAW264.7细胞iNOS和COX-2表达[J]. 口腔医学研究, 2023, 39(3): 236-241.
[13]	闫锐, 郭凯利, 曹丰弟, 张军, 马哲. 牙龈卟啉单胞菌GroEL对人脐静脉内皮细胞影响的研究[J]. 口腔医学研究, 2022, 38(9): 872-876.
[14]	谢元栋, 李泽华, 刘晓莉, 胡玲, 詹骆宁, 李毅. 牙龈卟啉单胞菌脂多糖刺激人牙髓干细胞炎症相关蛋白质组学研究[J]. 口腔医学研究, 2022, 38(8): 736-741.
[15]	高源, 李佳, 李维善. 牙龈卟啉单胞菌脂多糖对HUVECs中COX-2表达及信号通路的研究[J]. 口腔医学研究, 2022, 38(5): 436-441.