摘要
本文首次采用斑马鱼模型动物,以糖皮质激素地塞米松为模型药,以建立一种快速、高效的骨质疏松新模型。将受精后4天(4 dpf)的斑马鱼幼鱼暴露于不同浓度地塞米松溶液中,设阴性(0.5%DMSO)对照组,28.5℃下在24孔板中培养至9 dpf,至斑马鱼头部骨骼形成。并用此模型观察依替膦酸二钠对斑马鱼头骨矿化的影响及对10μmol.L 1地塞米松诱导的斑马鱼骨质疏松的防治作用。采用茜素红对培养9天的斑马鱼骨骼进行染色,以显微检测、数码成像方法定量分析骨骼染色区域。结果显示:与阴性对照组相比,地塞米松(2.5、10和25μmol.L 1)组的斑马鱼头部骨骼染色面积和染色光密度值显著减少;依替膦酸二钠(15及30μg.mL 1)能增加斑马鱼头骨矿化量并有效防止地塞米松诱导斑马鱼产生的骨质疏松。结果提示,地塞米松使斑马鱼头骨骼矿化量和骨密度显著降低,成功诱导斑马鱼产生骨质疏松。该模型亦成功验证依替膦酸二钠能防止骨质疏松的作用。
Zebrafish was selected as model animal, and glucocorticoid dexamethasone was used as a model compound to establish a rapid and high efficient osteopenia model. Zebrafish larvae at 4 days post fertilization （dpf） were exposed to a serial concentrations of dexamethasone solutions, and 0.5% DMSO was selected as the vehicle control group. All groups were incubated in 24-well plates （28.5 ℃） until 9 dpf. In addition, effects of 10 μmol·L？1 dexamethasone on preventing against osteopenia induced by etidronate disodium were also investigated. Zebrafish bones at 9 dpf were stained with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results showed that dexamethasone group at 2.5, 10 and 25 μmol·L？1 can decrease the staining area and the staining optical density values of zebrafish head bones when compared with the vehicle control group （0.5% DMSO）, which suggested that dexamethasone can significantly reduce the zebrafish mineralized bone and the bone mineral density. Results also showed that 15 and 30 μg·mL？1 etidronate disodium can increase the mineralized matrix of zebrafish head bone and prevent against osteopenia induced by dexamethasone. In conclusion, the study indicated that zebrafish can be an idea osteopenia model induced by dexamethasone.
出处
《药学学报》
CAS
CSCD
北大核心
2013年第2期255-260,共6页
Acta Pharmaceutica Sinica
基金
国家自然科学基金资助项目(30973978)
江苏省自然科学基金资助项目(BK2011866)
