Screening on antiosteoporotic active parts of Epimedium koreanum based on zebrafish model / 中草药

ABSTRACT

Objective: Prednisolone-induced osteoporosis model using zebrafish was used to screen the antiosteoporotic active parts of Epimedium koreanum, and to investigate the applicability and rationality of the zebrafish model of osteoporosis. Methods: Zebrafish larva at 4 d post fertilization were exposed with 25 μmol/L prednisolone (model group), 15 μg/mL etidronate disodium with 25 μmol/L prednisolone (positive group), 0.5% DMSO (vehicle control group), E. koreanum extracts (crude drug 0.1, 1.0, 10.0, and 100.0 μg /mL) with 25 μmol/L prednisolone, and their water and 30%, 50%, 70%, and 90% ethanol elution parts of macroporous resin (crude drug 1.0, 10.0, 100.0, and 1 000.0 μg/mL). All groups were incubated in 24-well plates (28.5°C) for 9 d until execution, and then zebrafish skeleton was 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 head skeleton mineral area and integrated optical density (IOD) of 25 μmol/L prednisolone model group were significantly decreased when compared with vehicle control group, and E. koreanum extract and its 30%, 50%, 70%, and 90% ethanol elution parts of macroporous resin could rescue the further bone loss of zebrafish induced by prednisolone when compared with model group. HPLC analysis indicated that the components of 30%, 50%, 70%, and 90% ethanol elution parts of macroporous resin mainly included flavonoids and minor nonflavonoids components. Conclusion: This novel osteoporosis zebrafish model is successfully used to screen the antiosteoporotic active parts of E. koreanum, which has the advantages of simplity, high efficiency, and easy to perform.