泼尼松龙诱导斑马鱼骨质疏松模型的建立

Zebrafish Model of Osteoporosis Induced by Prednisolone

本文首次采用斑马鱼为模型动物,以糖皮质激素泼尼松龙为造模用药,建立一种快速、高效的骨质疏松新动物模型。选用受精后4 d或5 d的幼鱼暴露于不同浓度泼尼松龙溶液中,同时设0.5%DMSO溶媒阴性对照组,28.5℃下在24孔板中培养5 d,至斑马鱼颅骨形成。采用茜素红对培养至9 d或10 d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。与溶媒阴性对照组相比,2.5,10,25μmol.L-1泼尼松龙组的斑马鱼骨骼染色面积和染色光密度值显著减少,提示泼尼松龙使斑马鱼骨骼矿化量和骨密度显著降低,成功诱导斑马鱼骨质疏松。并用该模型成功验证鲑鱼降钙素和依替膦酸二钠防治骨质疏松的作用。

Zebrafish was selected as model animal,and glucocorticoid prednisolone was used as a model compound to develop a rapid and high efficient osteoporosis novel model.A 5-day exposure was given to a series of prednisolone solutions with a range of concentration starting from either 4 days post fertilization（dpf） or 5 dpf.And 0.5% DMSO was selected as the negative solvent control group.All groups were incubated in 24-well plates（28.5℃）.Zebrafish at 9 dpf or 10 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.And the results showed that the prednisolone group at the concentration of 2.5 μmol.L-1,10 μmol.L-1 and 25 μmol.L-1 can obviously decrease the staining area and the staining optical density values when compared with the negative solvent control group（0.5% DMSO）,which suggested that prednisolone can significantly reduce the zebrafish mineralized bone and the bone mineral density.Therefore,osteoporosis model using zebrafish induced by prednisolone was successfully developed.In addition,zebrafish osteoporosis model was successfully used to verify the effect of salmon calcitonin and etidronate disodium in the prevention of osteoporosis.