Alteration of Osteocalcin MRNA Expression in Ovine Osteoblasts in Dependence of Sodium Fluoride and Sodium Selenite Medium Supplementation
Alteration of Osteocalcin MRNA Expression in Ovine Osteoblasts in Dependence of Sodium Fluoride and Sodium Selenite Medium Supplementation
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DOI：
10.1556/abiol.61.2010.1.6
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发表时间：
2010-03
影响因子：
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通讯作者：
Liting Sun;Fuqing Yu;Zhi-hong Xu;Xi-bo Zeng;M. Ferreri;B. Han
中科院分区：
文献类型：
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作者：
Liting Sun;Fuqing Yu;Zhi-hong Xu;Xi-bo Zeng;M. Ferreri;B. Han
本研究的目的是评估用不同浓度的氟化钠 (F) 和亚硒酸钠 (Se) 培养的绵羊成骨细胞对骨钙素 (OCN) 表达的定量，以评估这些试剂在体外对 OCN 表达的相互作用。我们想证明硒对氟化物的毒性作用可能具有保护作用。通过完全胰蛋白酶和胶原酶消化从绵羊颅骨中分离出成骨细胞，并在加湿的5%CO_2培养箱中于37℃下在补充有15%FBS的DMEM中培养。将鉴定出的成骨细胞分为1个对照组（C）和8个实验组，将其暴露于不同浓度的氟化钠（F；0、0.5、1 mM）和亚硒酸钠（Se；0、0.1、1 μM）。在处理后的不同时间点提取总RNA并反转录成第一链cDNA。通过实时荧光定量 PCR (qPCR) 间接测量 OCN mRNA。 F 1 mM with Se 1 μM 组 OCN mRNA 表达在第 7 天出现高峰，前后较低。除对照组外，所有组中 OCN mRNA 的表达均可被 F 和/或 Se 促进，表现出普遍上调。此外，通过使用中等浓度的硒可以避免成骨细胞过度暴露于氟而产生的毒性。体外培养的成骨细胞在最初几天可能会对 F 和 Se 产生应激反应。低浓度的Se可抑制高浓度F的毒性作用。因此，F和Se可作为拮抗因子，调节骨钙素的表达。
Objective of this study was to assess the quantification of osteocalcin (OCN) expression by ovine osteoblasts cultured with different concentrations of sodium fluoride (F) and sodium selenite (Se) to evaluate the interaction of these agents on OCN expression in vitro. We wanted to demonstrate a possible protective effect of selenium on the toxic effect of fluoride. Osteoblasts were isolated by complete trypsin and collagenase digestion from ovine calvarial bone and cultured in DMEM supplemented with 15% FBS at 37 °C in a humidified 5% CO_2 incubator. Identified osteoblasts were divided into one control group (C) and eight experimental groups, which were exposed to different concentrations of sodium fluoride (F; 0, 0.5, 1 mM) sodium selenite (Se; 0, 0.1, 1 μM). At different time points after treatment total RNA was extracted and reverse transcribed into first-strand cDNA. OCN mRNA was indirectly measured by real-time fluorescent quantitative PCR (qPCR). OCN mRNA expression in F 1 mM with Se 1 μM group was found to have a high peak at day seven and was lower before and afterwards. Expression of OCN mRNA in all groups except control could be promoted by F and/or Se showing a general upregulation. Furthermore, the toxicity from excessive exposure of osteoblast with F could be circumvented by usage of moderate concentration of Se. Osteoblasts cultured in vitro may have stressful responses to F and Se at the first few days. Low concentrations of Se inhibit the toxic effects of high concentrations of F. Therefore, F and Se could be used as antagonistic factors, which could regulate osteocalcin expression.