高グルコースによる歯肉線維芽細胞とTHP-1マクロファージのサイトカイン制御 : 糖尿病患者における歯周病重症化機序
High Glucose Induces Severe Periodontitis.
リュウ, チョン ファン 徳島大学大学院口腔科学教育部（口腔科学専攻）
成石, 浩司 Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
梶浦, 由加里 Tokushima University KAKEN研究者をさがす
西川, 泰史 Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
生田, 貴久 Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
木戸, 淳一 Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
永田, 俊彦 Tokushima University 徳島大学 教育研究者総覧 KAKEN研究者をさがす
Background/Aims: Diabetic patients are susceptible to severe periodontitis, but the precise mechanism is not fully understood. Aim of this study was to explore the biological pathogenesis of severe periodontitis in diabetic patients focusing on the crosstalk of human gingival fibroblasts (HGFs) and macrophages. Methods: A total of 70 periodontitis patients with or without diabetes mellitus (DM) were enrolled, and the statistical relationships of diabetic conditions to the periodontal inflammatory parameters were examined by cross-sectional study. In in vitro study, HGFs cell line CRL-2014® (ATCC) and differentiated THP-1 macrophages were cultured with normal glucose (NG: 5.5 mM) or high glucose (HG: 25 mM) condition, and treated with indicated inflammatory factors such as calprotectin (CPT), interleukin (IL)-1β and IL-6. To examine the effects of HG on soluble IL-6 receptor (sIL-6R) production in THP-1 macrophages, the supernatants were collected and the sIL-6R levels were measured by ELISA. To examine the effects of HG on IL-1β or IL-6-induced matrix metalloproteinase (MMPs) production in HGFs, the supernatants were collected. Levels of MMP-1 and tissue inhibitor of MMP-1 (TIMP-1) were measured by ELISA. Finally, after conditioned medium (CM) from THP-1 macrophages cultured with NG or HG conditions was collected, HGFs were treated with the CM. The supernatants were collected 24 hours later and the levels of MMP-1 and TIMP-1 were measured. To examine the specific effects of IL-1β contained in CM on MMP-1 and TIMP-1 production in HGFs, IL-1 receptor antagonist (IL-1ra) was used. Results: There were statistical correlation between IL-1β and sIL-6R levels in gingival crevicular fluid (GCF) and HbA1c in periodontitis patients with DM (IL-1β: P=0.035, sIL-6R: P=0.040). HG and CPT significantly induced sIL-6R production in THP-1 macrophages. HG significantly enhanced IL-1β or IL-6/sIL-6R-induced MMP-1 production in HGFs. The increase of MMP-1 by both IL-1β and IL-6/sIL-6R was significantly inhibited by specific ERK or IκB inhibitors. Corresponding to the regulation of MMP-1 production, HG condition increased the phosphorylation of p44/42 MAPK and IκBα in HGFs treated with IL-1β or IL-6/sIL-6R. Finally, MMP-1 production in HGFs cultured with HG increased significantly by CM from THP-1 macrophages cultured with HG. The induction of MMP-1 by the CM from THP-1 macrophages cultured with HG was significantly inhibited by dose dependent of IL-1ra in HGFs cultured with HG. Conclusion: Diabetic conditions such as HG induce IL-1β and sIL-6R production from macrophages in inflammatory periodontal tissues and may exacerbate the periodontitis synergistically via MMP-1 production from HGFs.
Cellular Physiology and Biochemistry
This article is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND) ( http://www.karger.com/Services/OpenAccessLicense ). Usage and distribution for commercial purposes as well as any distribution of modified material requires written permission.
k3280_abstract.pdf 150 KB
k3280_review.pdf 43 KB
k3280_fulltext.pdf 1.28 MB
k3280_supple.pdf 99.8 KB