Evaluation of antiosteoporotic activity for micro amount icariin and epimedin B based on the osteoporosis model using zebrafish / 中国药学杂志

ABSTRACT

OBJECTIVE: Prednisolone-induced osteoporosis model using zebrafish was used to evaluate the antiosteoporotic activity of micro amount icariin and epimedin B. METHODS: Zebrafish larvae were co-exposed with 25 μmol·L-1 prednisolone and a series of icariin and epimedin B solutions with a range of concentration (0.2, 1, 5 and 10 μmol·L-1) from 4 to 9 days post fertilization(dpf), 25 μmol·L-1 prednisolone was selected as model group, 15 μg·mL-1 etidronate disodium with 25 μmol·L-1 prednisolone 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°C) 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 skeleton mineral aera and integrated optical density (IOD) of 25 μmol·L-1 prednisolone model group were significantly decreased when compared with vehicle control group and negative control group; and 15 μg·mL-1 etidronate disodium can increase the mineralized matrix of zebrafish head skeleton when compared with model group and prevent osteoporosis induced by prednisolone; head skeleton mineral area and IOD of 10 and 5 μmol·L-1 icariin groups and 1μmol·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.