ABSTRACT
BACKGROUND: Our previous studies have found that Bushen Tiaogan prescription can significantly reduce pain and improve the clinical symptoms of traditional Chinese medicine in elderly patients with osteoporosis. The clinical effect is positive, but there is no relevant experimental evidence. OBJECTIVE: To explore the safety of Bushen Tiaogan prescription and its therapeutic effect on prednisolone-induced osteoporosis. METHODS: The zebrafish juveniles 5 hours after fertilization were evenly distributed into 24-well plates at a density of 20 per well. Different concentrations of Bushen Tiaogan prescription (crude drug concentrations 10, 50, 100, 250, 500 mg/L) were added to the corresponding wells. The development of the embryos was observed at 24, 48, 72, 96 hours after fertilization, and the cumulative survival rate and hatching rate were used to evaluate the safety of Bushen Tiaogan prescription. The zebrafish juveniles 3 days after fertilization were randomly divided into blank control group (0.5% dimethyl sulfoxide), 25 µmol/L prednisolone model group, alendronate sodium positive drug group and Bushen Tiaogan prescription (10, 50, 100, 250, 500 mg/L) groups, with 40 in each group. The culture fluid was changed every day and executed at the 8th day. Zebrafish juveniles were dyed using alizarin red for microscopic detection and digital imaging and the bone staining area was quantitatively analyzed by image software to evaluate the anti-osteoporosis activity of Bushen Tiaogan prescription. RESULTS AND CONCLUSION: The low-concentration Bushen Tiaogan prescription had no effect on embryo survival rate and hatching rate, while the high-concentration Bushen Tiaogan prescription could reduce embryo survival rate and hatching rate. Prednisolone, 25 µmol/L, was able to successfully induce significant loss of bone mass in zebrafish. Compared with the model group, 50, 100 and 250 mg/L Bushen Tiaogan prescription could resist prednisolone-induced zebrafish osteoporosis in a concentration-dependent manner. At low concentrations, there was no significant anti-osteoporosis effect, and at high concentrations, anti-osteoporosis effect showed a downward trend. These findings indicate that 25 µmol/L prednisolone successfully induces the establishment of zebrafish osteoporosis model, and successfully evaluates the initial safety of Bushen Tiaogan prescription. Bushen Tiaogan prescription begins to show an anti-osteoporosis effect at 50 mg/L, and has the most significant anti-osteoporosis effect at 250 mg/L.