Department of Periodontology, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases, Beijing, China.
Department of Periodontology, Peking University School and Hospital of Stomatology & National Center of Stomatology & National Clinical Research Center for Oral Diseases, Beijing, China.
Int Dent J. 2024 Jun;74(3):607-615. doi: 10.1016/j.identj.2023.12.006. Epub 2024 Jan 15.
Periodontitis is one of the most common chronic oral inflammatory diseases. Over the past decade, herpes viruses, particularly Epstein-Barr virus (EBV), have been considered promising pathogenic candidates for periodontitis. However, the specific mechanism by which EBV contributes to the development of periodontitis is still unknown. This study aimed to explore the mechanism of EBV underlying the inflammatory response in human gingival fibroblasts (HGFs).
HGFs were stimulated with different concentrations of EBV (10, 10, 10, 10, and 10 DNA copies/mL) for 0, 8, 24, or 48 hours. The mRNA levels of interleukin (IL)-1β, tumour necrosis factor-α (TNF-α), IL-8, monocyte chemoattractant protein-1 (MCP-1), and Toll-like receptor 9 (TLR9) were measured using quantitative real-time polymerase chain reaction (PCR). Enzyme-linked immunosorbent assays (ELISAs) were performed for determining the mRNA and protein levels of IL-1β, TNF-α, IL-8, and MCP-1. Real-time PCR and ELISA were performed to determine the protein levels of IL-1β, TNF-α, IL-8, and MCP-1. Activation of the TLR9/myeloid differentiation factor 88 (MyD88)/nuclear factor kappa B (NF-κB) pathway was evaluated using western blotting.
The expressions of IL-1β, TNF-α, IL-8, and MCP-1 were significantly upregulated in HGFs under EBV stimulation in a concentration- and time-dependent manner. EBV promoted TLR9 and MyD88 expression and induced NF-κB transcription. On the contrary, the upregulation of these factors and the activation of NF-κB pathway were drastically inhibited by TLR9 antagonists.
Our findings demonstrate that EBV promotes the production of inflammatory cytokines IL-1β and TNF-α and chemokines IL-8 and MCP-1 in HGFs through the TLR9/MyD88/NF-κB pathway.
牙周炎是最常见的慢性口腔炎症性疾病之一。在过去的十年中，疱疹病毒，特别是 Epstein-Barr 病毒（EBV），已被认为是牙周炎有希望的致病候选物。然而，EBV 促进牙周炎发展的确切机制尚不清楚。本研究旨在探讨 EBV 引起人牙龈成纤维细胞（HGFs）炎症反应的机制。
用不同浓度的 EBV（10、10、10、10 和 10 DNA 拷贝/ml）刺激 HGFs 0、8、24 或 48 小时。用实时定量聚合酶链反应（PCR）检测白细胞介素（IL）-1β、肿瘤坏死因子-α（TNF-α）、IL-8、单核细胞趋化蛋白-1（MCP-1）和 Toll 样受体 9（TLR9）的 mRNA 水平。用酶联免疫吸附试验（ELISA）测定 IL-1β、TNF-α、IL-8 和 MCP-1 的 mRNA 和蛋白水平。用实时 PCR 和 ELISA 测定 IL-1β、TNF-α、IL-8 和 MCP-1 的蛋白水平。用 Western blot 法测定 TLR9/髓样分化因子 88（MyD88）/核因子 kappa B（NF-κB）通路的激活情况。
EBV 刺激 HGFs 后，IL-1β、TNF-α、IL-8 和 MCP-1 的表达呈浓度和时间依赖性显著上调。EBV 促进 TLR9 和 MyD88 的表达，并诱导 NF-κB 转录。相反，TLR9 拮抗剂可显著抑制这些因子的上调和 NF-κB 通路的激活。
我们的研究结果表明，EBV 通过 TLR9/MyD88/NF-κB 通路促进 HGFs 中炎症细胞因子 IL-1β 和 TNF-α以及趋化因子 IL-8 和 MCP-1 的产生。
