Expression of glutathione peroxidase 1 in the spheno-occipital synchondrosis and its role in ROS-induced apoptosis

Eur J Orthod. 2015 Jun;37(3):308-13. doi: 10.1093/ejo/cju045. Epub 2014 Oct 13.

Abstract

Background/objective: Chondrogenesis is an integral part of endochondral bone formation, by which the midline cranial base is developed. Reactive oxygen species (ROS) are required in chondrogenic differentiation and antioxidant enzymes regulate their levels. The aim of this study was to localize the antioxidant enzyme glutathione peroxidase 1 (Gpx1) at the spheno-occipital synchondrosis, as well as its effect on ROS challenge and its expression pattern in the course of differentiation.

Materials and methods: Gpx1 was semiquantified in immunohistochemically stained sections of spheno-occipital synchondroses of rats. The effect of Gpx1 on ROS-induced apoptosis was investigated by manipulating the expression of Gpx1 in ATDC5 cells. The temporal pattern of Gpx1 expression was determined during chondrocyte differentiation for 21 days in vitro.

Results: Proliferating chondrocytes exhibited the greatest Gpx1 immunoreactivity and hypertrophic ones the lowest (P = 0.02). Cells transfected with Gpx1-siRNA had the highest apoptotic rate, while cells overexpressing Gpx1 the lowest one (P < 0.001). Gpx1 was significantly increased on days 10 (P = 0.02) and 14 (P = 0.01).

Conclusions: Hypertrophic chondrocytes have the lowest Gpx1 activity in the spheno-occipital synchondrosis. Gpx1 is implicated in the ROS-induced apoptosis in chondrocytes. Its expression was not constitutive during chondrogenic differentiation.

MeSH terms

  • Animals
  • Animals, Newborn
  • Apoptosis / physiology*
  • Cell Culture Techniques
  • Cell Differentiation / physiology
  • Cell Line
  • Cell Proliferation
  • Chondrocytes / enzymology
  • Chondrocytes / physiology
  • Chondrogenesis / physiology
  • Cranial Sutures / cytology
  • Cranial Sutures / enzymology*
  • Gene Knockdown Techniques
  • Glutathione Peroxidase / analysis*
  • Glutathione Peroxidase / genetics
  • Glutathione Peroxidase / physiology
  • Glutathione Peroxidase GPX1
  • Hypertrophy
  • Occipital Bone / cytology
  • Occipital Bone / enzymology*
  • Osteogenesis / physiology
  • RNA, Small Interfering / genetics
  • Rats
  • Rats, Wistar
  • Reactive Oxygen Species / analysis*
  • Reactive Oxygen Species / pharmacology
  • Skull Base / cytology
  • Sphenoid Bone / cytology
  • Sphenoid Bone / enzymology*
  • Time Factors

Substances

  • RNA, Small Interfering
  • Reactive Oxygen Species
  • Glutathione Peroxidase
  • Glutathione Peroxidase GPX1
  • Gpx1 protein, rat