Department of Periodontology, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, Jinan, Shandong, China.
Department of General Dentistry, School and Hospital of Stomatology, Cheeloo College of Medicine, Shandong University & Shandong Key Laboratory of Oral Tissue Regeneration & Shandong Engineering Research Center of Dental Materials and Oral Tissue Regeneration & Shandong Provincial Clinical Research Center for Oral Diseases, Jinan, Shandong, China.
Oral Dis. 2024 Oct;30(7):4767-4781. doi: 10.1111/odi.14899. Epub 2024 Feb 26.
This study aimed to clarify the difference in Fusobacterium nucleatum (F. nucleatum) induced inflammatory cytokines and nod-like receptor protein 3 (NLRP3) inflammasomes dysregulation among three periodontal cells.
Oral epithelial cells (HIOECs), THP-1 macrophages, and human gingival fibroblasts (HGFs) were exposed to F. nucleatum with/without adenosine triphosphate (ATP) and nigericin (Nig). Cell morphology was assessed by scanning electron microscopy. qRT-PCR, protein microarrays, and bioinformatic methods were used to evaluate the cytokines and their complex interplay. NLRP3 inflammasomes activation was detected by western blotting and ELISA.
F. nucleatum adhered to and invaded cells. In HIOECs, F. nucleatum enhanced interleukin (IL)-1α/1β/6/10/13, TNF-α, and interferon (IFN)-γ expression. In THP-1 macrophages, F. nucleatum up-regulated IL-1α/1β/6/10 and TNF-α levels. In HGFs, F. nucleatum increased IL-6 levels. F. nucleatum and ATP synergistically boosted IFN-γ level in THP-1 macrophages and IL-13 level in HGFs. IL-1α/1β/6, and TNF-α served as epicenters of the inflammatory response. Additionally, F. nucleatum activated NLRP3 inflammasomes in HIOECs, and ATP/Nig boosted the activation. F. nucleatum also triggered NLRP3 inflammasomes in THP-1 macrophages, but in HGFs, only NLRP3 and caspase-1 levels were elevated.
F. nucleatum infiltrated periodontal supporting cells and dysregulated inflammatory cytokines and NLRP3 inflammasomes.
本研究旨在阐明三种牙周细胞中，具核梭杆菌（Fusobacterium nucleatum，F. nucleatum）诱导的炎症细胞因子和核苷酸结合寡聚化结构域样受体蛋白 3（NLRP3）炎性小体失调的差异。
将口腔上皮细胞（HIOECs）、THP-1 巨噬细胞和人牙龈成纤维细胞（HGFs）分别用含/不含三磷酸腺苷（adenosine triphosphate，ATP）和 Nigericin（Nig）的具核梭杆菌进行孵育。通过扫描电子显微镜评估细胞形态。采用 qRT-PCR、蛋白质微阵列和生物信息学方法评估细胞因子及其相互作用。通过 Western blot 和 ELISA 检测 NLRP3 炎性小体的激活。
F. nucleatum 黏附和侵袭细胞。在 HIOECs 中，F. nucleatum 增强白细胞介素（interleukin，IL）-1α/1β/6/10/13、肿瘤坏死因子（tumor necrosis factor，TNF）-α 和干扰素（interferon，IFN）-γ 的表达。在 THP-1 巨噬细胞中，F. nucleatum 上调了 IL-1α/1β/6/10 和 TNF-α 水平。在 HGFs 中，F. nucleatum 增加了 IL-6 水平。F. nucleatum 和 ATP 协同增强了 THP-1 巨噬细胞中的 IFN-γ 水平和 HGFs 中的 IL-13 水平。IL-1α/1β/6、TNF-α 是炎症反应的中心。此外，F. nucleatum 激活了 HIOECs 中的 NLRP3 炎性小体，ATP/Nig 增强了其激活。F. nucleatum 还触发了 THP-1 巨噬细胞中的 NLRP3 炎性小体，但在 HGFs 中，只有 NLRP3 和半胱天冬酶-1（caspase-1）水平升高。
F. nucleatum 侵袭牙周支持细胞，扰乱炎症细胞因子和 NLRP3 炎性小体。
