Characterization of diabetic osteoarthritic cartilage and role of high glucose environment on chondrocyte activation: toward pathophysiological delineation of diabetes mellitus-related osteoarthritis

Osteoarthritis Cartilage. 2015 Sep;23(9):1513-22. doi: 10.1016/j.joca.2015.04.026. Epub 2015 May 15.

Abstract

Objective: To examine the relationship between osteoarthritis (OA) and type 2 diabetes mellitus (DM).

Methods: OA cartilage from DM and non-DM patients undergoing knee replacement were stimulated by IL-1β for 24 h and release of interleukin-6 (IL-6) and prostaglandin E2 (PGE2) was measured. Primary cultured murine chondrocytes were stimulated for 24 and 72 h with or without IL-1β (5 ng/mL) under normal-glucose (5.5 mM) or high-glucose (25 mM) conditions. The expression and release of pro-inflammatory mediators (IL-6, cyclooxygenase 2 [COX2]/PGE2) were analyzed by quantitative RT-PCR and ELISA/EIA. Glucose uptake was assessed with ((14)C)-2-deoxyglucose. Reactive oxygen species (ROS) and nitric oxide (NO) production were measured. To analyze the mechanism of IL-1β-induced inflammation, cells were pretreated or treated with inhibitors of glucose transport (cytochalasin B), the polyol pathway (epalrestat), mitochondrial oxidative stress (MitoTEMPO) or nitric oxide synthase (l-NAME).

Results: With IL-1β stimulation, IL-6 and PGE2 release was greater in human DM than non-DM OA cartilage (2.7- and 3-fold, respectively) (P < 0.05). In vitro, with IL-1β stimulation, IL-6 and COX2 mRNA expression, IL-6 and PGE2 release, and ROS and NO production were greater under high-than normal-glucose conditions in cultured chondrocytes. IL-1β-increased IL-6 release was reduced with cytochalasin B, epalrestat, L-NAME or MitoTEMPO treatment (-45%, -62%, -38% and -40%, respectively).

Conclusion: OA cartilages from DM patients showed increased responsiveness to IL-1β-induced inflammation. Accordingly, high glucose enhanced IL-1β-induced inflammation in cultured chondrocytes via oxidative stress and the polyol pathway. High glucose and diabetes may thus participate in the increased inflammation in OA.

Keywords: Diabetes mellitus; Glucose; Metabolic osteoarthritis; Osteoarthritis; Oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cartilage / metabolism*
  • Cells, Cultured
  • Cyclooxygenase 2 / metabolism
  • Diabetes Mellitus, Type 2 / complications*
  • Dinoprostone / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Glucose / metabolism
  • Glucose / physiology
  • Humans
  • Interleukin-1beta / physiology
  • Interleukin-6 / metabolism
  • Male
  • Middle Aged
  • Osteoarthritis / etiology*
  • Osteoarthritis / physiopathology
  • Oxidative Stress / physiology
  • Peptide Fragments / physiology
  • Polymers / metabolism
  • Real-Time Polymerase Chain Reaction

Substances

  • Interleukin-1beta
  • Interleukin-6
  • Peptide Fragments
  • Polymers
  • polyol
  • interleukin-1beta (163-171)
  • Cyclooxygenase 2
  • Glucose
  • Dinoprostone