Koretsi Vasiliki, Kirschneck Christian, Proff Peter, Römer Piero
Department of Orthodontics, University Medical Centre Regensburg, Germany.
Department of Orthodontics, University Medical Centre Regensburg, Germany
Eur J Orthod. 2015 Jun;37(3):308-13. doi: 10.1093/ejo/cju045. Epub 2014 Oct 13.
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.
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.
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).
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.
背景/目的:软骨形成是软骨内骨形成的一个组成部分,通过软骨内骨形成发育中线颅底。软骨分化过程中需要活性氧(ROS),抗氧化酶可调节其水平。本研究的目的是在蝶枕软骨结合处定位抗氧化酶谷胱甘肽过氧化物酶1(Gpx1),以及其对ROS刺激的影响及其在分化过程中的表达模式。
在大鼠蝶枕软骨结合处的免疫组织化学染色切片中对Gpx1进行半定量分析。通过操纵ATDC5细胞中Gpx1的表达来研究Gpx1对ROS诱导的细胞凋亡的影响。在体外软骨细胞分化21天的过程中确定Gpx1表达的时间模式。
增殖期软骨细胞表现出最强的Gpx1免疫反应性,肥大期软骨细胞表现出最弱的Gpx1免疫反应性(P = 0.02)。转染Gpx1-siRNA的细胞凋亡率最高,而过度表达Gpx1的细胞凋亡率最低(P < 0.001)。在第10天(P = 0.02)和第14天(P = 0.01)Gpx1显著增加。
在蝶枕软骨结合处,肥大期软骨细胞的Gpx1活性最低。Gpx1与软骨细胞中ROS诱导的细胞凋亡有关。在软骨分化过程中其表达不是组成性的。