以二维斑马鱼模型联合色谱技术筛选朝藿定A及其代谢物宝藿苷I的抗骨质疏松活性,建立高效筛选中药抗骨质疏松活性成分新方法。用斑马鱼成鱼对朝藿定A的0.5%DMSO溶液进行代谢,代谢液浓缩后用LC-MS分析、HPLC捕获代谢产物宝藿苷I,用斑马鱼骨质疏松模型评价朝藿定A和宝藿苷I的活性。结果表明朝藿定A和宝藿苷I均能防止泼尼松龙诱导斑马鱼骨质疏松。该方法使朝藿定A的量微代谢产物得到富集、分离分析,通过在体斑马鱼抗骨质疏松模型简单、高效的筛选朝藿定A及代谢产物宝藿苷I的抗骨质疏松活性,为早期、快速发现中药量微成分及其代谢物的抗骨质疏松活性提供新思路与方法。
This article firstly established a new efficient method for screening anti-osteoporosis ingredients, which used two-dimensional zebrafish model combined with hyphenated chromatographic techniques to evaluate anti-osteoporosis activities of epimedin A and its metabolite baohuoside I. Adult zebrafish was used for metabolism of epimedin A in 0.5% DMSO, and LC-MS was used for analysis of the metabolite, which was captured by HPLC, and prednisolone-induced osteoporosis model of zebrafish was used to evaluate the anti-osteoporotic activities of trace amounts of epimedin A and baohuoside I. The results indicated that epimedin A and baohuoside I can prevent prednisolone-induced osteoporosis in zebrafish. The developed method in this paper enables the separation, enrichment and analysis of micro-amount metabolite of epimedin A, and anti-osteoporosis activities in vivo of epimedin A and baohuoside I was simple and efficient screening resorting to zebrafish osteoporosis mode. This paper would provide new ideas and methods for a rapid and early discovery of anti-osteoporosis activities of micro-ingredients and its metabolite of traditional Chinese medicine.