Evaluation of metabolism-effect/toxicity of Epimedium sagittatum based on zebrafish M-Act/Tox integration method / 中草药

ABSTRACT

Objective To evaluate the metabolism, anti-osteoporosis efficacy and toxicity of Epimedium sagittatum based on zebrafish M-Act/Tox integration method. Methods The safety of E. sagittatum (crude drug 200, 500, 1 000, 1 500, and 2 000 μg/mL) was evaluated with 1-6 dpf zebrafish, the morphology of fish organs was observed and the number of deaths was counted and the half death concentration of zebrafish (LC50) was calculated; The 5 dpf zebrafish were exposed to E. sagittatum (crude drug 250 μg/mL ) for 24 h, six representative flavonoids, including epimedin A, epimedin B, epimedin C, icariin, sagittatoside C, and baohuoside I, were analyzed; The zebrafish osteoporosis model was induced with 25 μmol/L prednisolone, microscopic detection and digital imaging of zebrafish larvae of each group cultured to 8 dpf were performed using alizarin red, and the bone staining area was quantitatively analyzed by image software to evaluate the anti-osteoporosis activity of E. sagittatum. Results E. sagittatum caused zebrafish poisoning at crude drug 500 μg/mL and above concentration (organs deformation or death), and toxicity was related to drug concentration and exposing time; After the action of zebrafish, epimedin A, epimedin B, epimedin C, and icariin were almost completely transformed, and sagittatoside C and baohuoside I were the main metabolites; E. sagittatum had significant anti-prednisolone-induced bone loss in zebrafish at a certain concentration (crude drug 6.25, 12.5, and 25 μg/mL). Conclusion The metabolism, anti-osteoporosis efficacy and toxicity of E. sagittatum are evaluated using zebrafish. The zebrafish M-Act/Tox integration method integrates the advantages of zebrafish metabolism, osteoporosis model and toxicity evaluation method, and realizes the effect/toxicity evaluation based on in vivo process, providing new ideas and method for screening anti-osteoporosis Chinese medicine.