补肾运脾方对糖皮质激素性骨质疏松斑马鱼模型骨代谢的调节作用

Regulation of Bushen Yunpi Prescription on bone metabolism in zebrafish model with glucocorticoid-induced osteoporosis

目的:探究补肾运脾方对糖皮质激素性骨质疏松斑马鱼骨代谢的作用机制。方法:将受精后72h的斑马鱼幼鱼随机置于空白对照组、25μg/ml泼尼松龙造模组、阳性对照依替膦酸二钠300μg/mL组、补肾运脾方25、50、100μg/mL组,每组30条。每24 h记录斑马鱼的死亡率,144 h后进行麻醉,用0.2%钙黄绿素进行荧光染色,体视荧光显微镜下观察,拍照记录并计算斑马鱼脊椎骨的个数,脊椎硬骨染色面积(Area)和前三节椎体的荧光强度(S)。结果:与空白对照组相比,泼尼松龙处理的模型对照组的斑马鱼椎体个数、染色面积及荧光强度均明显降低;与模型对照组相比,100μg/mL补肾运脾方能显著增加斑马鱼椎体个数、染色面积及荧光强度。结论:25μg/mL泼尼松龙能成功诱导斑马鱼骨质疏松的建立,100μg/mL补肾运脾方能有效改善斑马鱼骨质疏松模型骨代谢并防治骨质疏松。

Objective: To explore the mechanism of Bushen Yunpi Prescription(BSYP) on bone metabolism in zebrafish with glucocorticoid-induced osteoporosis. Methods: Zebrafish embryos 72 h after fertilization were randomly placed in the control group, 25 ug/mL prednisolone model group, disodium etiphosphate(300 ug/mL) positive control group and different concentrations of Bushen Yunpi Prescription groups(25、50、100 ug/mL), 30 in each group. The mortality was recorded every 24 hours. 14 h after anesthesia, zebrafish were dyed by 0.2% calcein and observed under stereoscopic fluorescence microscope. Then vertebrae numbers, staining Area of spinal hard bones(Area) and fluorescence intensity of the first three vertebrae(S) were photographed and recorded.Results: Compared with the control group, the number of vertebral bodies, staining area and fluorescence intensity in prednisolone-treated model control group were significantly reduced. Compared with the model group, 100 ug/mL Bushen Yunpi Prescription significantly increased the number of zebrafish vertebral bodies, staining area and fluorescence intensity. Conclusion: 25 ug/mL prednisolone can successfully induce the establishment of zebrafish osteoporosis. 100 ug/mL Bushen Yunpi Prescription can effectively improve the bone metabolism of zebrafish osteoporosis and prevent osteoporosis.