High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes

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...

Full description

Bibliographic Details
Main Authors: Jung-Hwan Lew, Koji Naruishi, Yukari Kajiura, Yasufumi Nishikawa, Takahisa Ikuta, Jun-ichi Kido, Toshihiko Nagata
Format: Article
Language:English
Published: Cell Physiol Biochem Press GmbH & Co KG 2018-10-01
Series:Cellular Physiology and Biochemistry
Subjects:
Online Access:https://www.karger.com/Article/FullText/494481
id doaj-c87a03d1e22b4627a30ab56b8456cd96
record_format Article
spelling doaj-c87a03d1e22b4627a30ab56b8456cd962020-11-25T01:34:23ZengCell Physiol Biochem Press GmbH & Co KGCellular Physiology and Biochemistry1015-89871421-97782018-10-0150397398610.1159/000494481494481High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with DiabetesJung-Hwan LewKoji NaruishiYukari KajiuraYasufumi NishikawaTakahisa IkutaJun-ichi KidoToshihiko NagataBackground/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.https://www.karger.com/Article/FullText/494481DiabetesFibroblastsIL-1βMacrophagesPeriodontitisSIL-6R
collection DOAJ
language English
format Article
sources DOAJ
author Jung-Hwan Lew
Koji Naruishi
Yukari Kajiura
Yasufumi Nishikawa
Takahisa Ikuta
Jun-ichi Kido
Toshihiko Nagata
spellingShingle Jung-Hwan Lew
Koji Naruishi
Yukari Kajiura
Yasufumi Nishikawa
Takahisa Ikuta
Jun-ichi Kido
Toshihiko Nagata
High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes
Cellular Physiology and Biochemistry
Diabetes
Fibroblasts
IL-1β
Macrophages
Periodontitis
SIL-6R
author_facet Jung-Hwan Lew
Koji Naruishi
Yukari Kajiura
Yasufumi Nishikawa
Takahisa Ikuta
Jun-ichi Kido
Toshihiko Nagata
author_sort Jung-Hwan Lew
title High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes
title_short High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes
title_full High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes
title_fullStr High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes
title_full_unstemmed High Glucose-Mediated Cytokine Regulation in Gingival Fibroblasts and THP-1 Macrophage: a Possible Mechanism of Severe Periodontitis with Diabetes
title_sort high glucose-mediated cytokine regulation in gingival fibroblasts and thp-1 macrophage: a possible mechanism of severe periodontitis with diabetes
publisher Cell Physiol Biochem Press GmbH & Co KG
series Cellular Physiology and Biochemistry
issn 1015-8987
1421-9778
publishDate 2018-10-01
description 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.
topic Diabetes
Fibroblasts
IL-1β
Macrophages
Periodontitis
SIL-6R
url https://www.karger.com/Article/FullText/494481
work_keys_str_mv AT junghwanlew highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
AT kojinaruishi highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
AT yukarikajiura highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
AT yasufuminishikawa highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
AT takahisaikuta highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
AT junichikido highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
AT toshihikonagata highglucosemediatedcytokineregulationingingivalfibroblastsandthp1macrophageapossiblemechanismofsevereperiodontitiswithdiabetes
_version_ 1725072496213884928