基于两种饮食的斑马鱼骨质疏松模型评价骨肽的抗骨质疏松作用

Evaluation of the anti-osteoporosis effect of osteo peptide with two zebrafish osteoporosis models

试验评价骨肽的抗骨质疏松作用,通过优化传统的斑马鱼骨质疏松模型,为筛选促钙吸收的药物提供更适宜的斑马鱼骨质疏松模型。选择3 dpf发育正常的斑马鱼胚胎,用10μg/mL泼尼松诱导建立斑马鱼骨质疏松症模型,评价(250、500、1000、2000μg/mL)骨肽抗骨质疏松作用,同时增设低钙饮食的斑马鱼骨质疏松模型评价(250、500、1000、2000μg/mL)骨肽的抗骨质疏松作用,为减少低钙饮食对骨质疏松斑马鱼模型的影响,同时增设给予低钙饮食的模型组。干预4天后,采用0.2%钙黄绿素染色,在荧光显微镜下拍照,计算斑马鱼的脊椎骨荧光强度。结果表明,采用常规斑马鱼骨质疏松模型评价骨肽无抗骨质疏松作用;然而采用低钙饮食的斑马鱼骨质疏松模型评价骨肽与给予低钙饮食的模型组相比脊椎骨荧光强度显著减少。低钙饮食的斑马鱼骨质疏松模型建立成功,1000μg/mL骨肽能够增加斑马鱼脊椎骨的矿化量,并有效防止泼尼松诱导斑马鱼产生的骨质疏松。

To evaluate the anti-osteoporosis effect of osteo peptide,the traditional zebrafish osteoporosis model was optimized to provide a more appropriate zebrafish osteoporosis model for screening calcium absorption promoting drugs.Normal 3 dpf zebrafish embryos were selected and treated with 10μg/mL prednisone to establish a zebrafish osteoporosis model,the anti-osteoporosis effects of osteo peptide with 250,500,1000,2000μg/mL were evaluated.At the same time,the zebrafish osteoporosis model evaluation with low-calcium diet(250,500,1000,2000μg/mL)was added to the anti-osteoporosis effect of bone meal.in order to reduce the effect of low-calcium diet on osteoporotic zebrafish,the control group with a low-calcium diet was added.After 4 days of intervention,these zebrafish were dyed for 2 hours by 0.2%calcein,and then the fiuorescence intensity of vertebrae of zebrafish were observed,photographed and calculated under a stereoscopic fiuorescence microscope.The osteo peptide withconventional zebrafish osteoporosis model had no anti-osteoporosis effect;however,the fiuorescence intensity was significantly reduced with the zebrafish osteoporosis model with a low-calcium diet and the control group model given a low-calcium diet.The osteoporosis model of zebrafish with low calcium diet was successfully established.Osteo peptide at 1000μg/m L can increase the mineralization of zebrafish vertebrae and effectively prevent osteoporosis induced by prednisolone in zebrafish.