Preventive Effect of Jie-Gu-Tang on Osteoporosis Model Using Zebrafish Induced by Prednisolone / 世界科学技术-中医药现代化

ABSTRACT

Prednisolone-induced zebrafish osteoporosis model was used to explore the bone-strengthening effect of Jie-Gu-Tang (JGT). Zebrafish larvae of 5 days post fertilization (d.p.f.) were co-exposed with 25 μmol·L-1 pred-nisolone and a series of JGT solutions with a range of concentrations (0.025, 0.25, 2.5, 25 and 100 mg crude herb per liter). The 25 μmol·L-1 prednisolone was selected as the model group. Etidronate disodium (15 and 30 mg·mL-1) with 25 μmol·L-1 prednisolone was used as the positive group. And 0.5% DMSO was used as the vehicle control group. All groups were incubated in 24-well plates (28.5℃) until 10 d.p.f. Zebrafish skeleton at 10 d.p.f. was anes-thetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscop-ic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. The results showed that prednisolone group at 25 μmol·L-1 concentration can obviously decrease the staining area and the stain-ing optical density values when compared with the vehicle control group (0.5% DMSO). Compared with the model group, both etidronate disodium (15 and 30 mg·mL-1) and JGT (2.5, 25 and 100 mg crude herb per liter) can in-crease the mineralized matrix and integrated optical density (IOD) of zebrafish head skeleton significantly with dose-effect relationship. It was concluded that zebrafish osteoporosis model was successfully used in the evaluation on bone loss prevention and bone formation promotion of JGT, which provided basis for the reliability and reasonability of zebrafish model.