摘要
目的本实验利用泼尼松龙诱导的斑马鱼骨质疏松模型评价微量淫羊藿苷和朝藿定B抗斑马鱼骨质疏松的作用。方法将受精后4 d的斑马鱼幼鱼暴露在含25μmol·L-1泼尼松龙的不同浓度淫羊藿苷(0.2、1、5和10μmol·L-1)和朝藿定B(0.2、1、5和10μmol·L-1),同时设25μmol·L-1泼尼松龙模型药物组,含25μmol·L-1泼尼松龙的15μg·mL-1依替膦酸二钠阳性药物组,0.5%二甲基亚砜溶媒对照组和培养基阴性对照组。28.5℃下在24孔板中培养,每天换液至第9天处死。采用茜素红对培养9 d的各组斑马鱼幼鱼骨骼染色,并以显微检测、数码成像方法定量分析骨骼染色区域。结果与二甲基亚砜溶媒和培养基阴性对照组相比,25μmol·L-1泼尼松龙模型组的斑马鱼头部骨骼染色面积和染色光密度值显著减少;与模型组比较,15μg·mL-1依替膦酸二钠能增加斑马鱼头骨矿化量并有效防止泼尼松龙诱导斑马鱼产生的骨质疏松;与模型组比较,10和5μmol·L-1的淫羊藿苷和1μmol·L-1朝藿定B都能显著提高斑马鱼头部骨骼的染色面积和染色累积光密度值,提示淫羊藿苷和朝藿定B能防止泼尼松龙诱导的斑马鱼骨质疏松。结论斑马鱼骨质疏松模型具简单、高效、化合物用量少及可操作性强的优势,成功用于评价微量淫羊藿苷和朝藿定B的抗骨质疏松活性。
OBJECTIVE Prednisolone-induced osteoporosis model using zebrafish was used to evaluate the antiosteoporotic activ- ity of micro amount icariin and epimedin B. METHODS Zebrafish larvae were co-exposed with 25μmol·L-1 prednisolone and a series of icariin and epimedin B solutions with a range of concentration （0. 2, 1, 5 and 10μmol·L-1） from 4 to 9 days post fertiliza- tion （dpf）, 25 μmol·L-1 prednisnlone was selected as model group, 15 μmol·L-1etidronate disodium with 25 μmol·L-1 prednisohme as positive group, 0. 5% DMSO as the vehicle control group, the medium as a negative blank control group, all groups were incubated in 24-well plates （ 28.5℃ ） until 9 dpf. The medium/solution was changed half every day. Zebrafish skeleton at 9 dpf were anesthetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. RESULTS The results indicated head skele- ton mineral aera and integrated optical density （ IOD ） of 25 μmol·L-1 prednisolone model group were significantly deereased when compared with vehicle control group and negative control group; and 15 μmol·L-1 etidronate disodium can increase the mineralized matrix of zebrafish head skeleton when compared with model group and prevent osteoporosis indueed by prednisolone; head skeleton mineral aera and IOD of 10 and 5 μmol·L-1 ieariin groups and 1 μmol·L-1 epimedin B groups were significantly increased when compared with model group, which indicated that icariin and epimedin B can prevent zebrafish osteoporosis induced by prednisolone. CONCLUSION This novel osteoporosis model using zebrafish has advantages of simple, high efficiency, far less amount of compound needed and easy to perform, and it was successfully applied in quickly evaluating the antiosteoporosis activity of micro amount compounds such as icariin and epimedin B.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2014年第1期30-35,共6页
Chinese Pharmaceutical Journal
基金
国家自然科学基金资助项目(81274088
30973978)
江苏省自然科学基金资助项目(BK2011866)
江苏省"六大人才高峰"项目(YY006)
