Regulation of ghrelin receptor by microbial and inflammatory signals in human osteoblasts

Braz Oral Res. 2019 Apr 25:33:e025. doi: 10.1590/1807-3107bor-2019.vol33.0025.

Abstract

Recently, it has been suggested that the anti-inflammatory hormone ghrelin (GHRL) and its receptor GHS-R may play a pivotal role in periodontal health and diseases. However, their exact regulation and effects in periodontitis are not known. The aim of this in-vitro study was to investigate the effect of microbial and inflammatory insults on the GHS-R1a expression in human osteoblast-like cells. MG-63 cells were exposed to interleukin (IL)-1β and Fusobacterium nucleatum in the presence and absence of GHRL for up to 2 d. Subsequently, gene expressions of GHS-R1a, inflammatory mediators and matrix metalloproteinase were analyzed by real-time PCR. GHS-R protein synthesis and NF-κB p65 nuclear translocation were assessed by immunocytochemistry and immunofluorescence microscopy, respectively. IL-1β and F. nucleatum caused a significant upregulation of GHS-R1a expression and an increase in GHS-R1a protein. Pre-incubation with a MEK1/2 inhibitor diminished the IL-1β-induced GHS-R1a upregulation. IL-1β and F. nucleatum also enhanced the expressions of cyclooxygenase 2, CC-chemokine ligand 2, IL-6, IL-8, and matrix metalloproteinase 1, but these stimulatory effects were counteracted by GHRL. By contrast, the stimulatory actions of IL-1β and F. nucleatum on the GHS-R1a expression were further enhanced by GHRL. Our study provides original evidence that IL-1β and F. nucleatum regulate the GHS-R/GHRL system in osteoblast-like cells. Furthermore, we demonstrate for the first time that the proinflammatory and proteolytic actions of IL-1β and F. nucleatum on osteoblast-like cells are inhibited by GHRL. Our study suggests that microbial and inflammatory insults upregulate GHS-R1a, which may represent a protective negative feedback mechanism in human bone.

Publication types

  • Evaluation Study

MeSH terms

  • Analysis of Variance
  • Cells, Cultured
  • Fusobacterium nucleatum / physiology*
  • Humans
  • Immunohistochemistry
  • Interleukin-1beta / pharmacology*
  • Microscopy, Fluorescence
  • Osteoblasts / chemistry*
  • Osteoblasts / drug effects
  • Osteoblasts / microbiology
  • Periodontitis / microbiology
  • Periodontitis / pathology
  • Real-Time Polymerase Chain Reaction
  • Receptors, Ghrelin / analysis*
  • Receptors, Ghrelin / physiology
  • Statistics, Nonparametric
  • Up-Regulation / physiology

Substances

  • Interleukin-1beta
  • Receptors, Ghrelin