Bioactive Study on Mixtures of Epimedin C and Icariin Monomers with Invariant Molarity to Zebrafish Osteoporosis Model

Abstract

ObjectiveTo evaluate the anti-osteoporosis effect of the mixtures of epimedin C and icariin monomers with invariant molarity on zebrafish osteoporosis model. MethodsThe zebrafishes, fertilized 4 d ago, were exposed in 11 groups of nutrient solutions with prednisolone (25 μmol/L) as well as epimedin C and icariin (15 μmol/L) of various contents. The ratio of epimedin C and icariin in the 11groups were as follows: A (10:0), B (9:1), C (8:2), D (7:3), E (6:4), F (5:5), G (4:6), H (3:7), I (2:8), J (1:9), and K (0:10). Meanwhile, a negative control group with prednisolone (25 μmol/L) was prepare as S. The selected zebrafish fetus was put into the 24-hole culture plate, and ensure every 5 zebrafishes in 1 hole and 2 holes as a group. They were placed in incubator at 28.5 °C, and the daily changes of fluid were investigated until they were put to death on day 8 and then fixed. After dyeing with alizarin red, the segmental venter of zebrafish skulls was observed and quantitative analysis of dyed area was conducted. ResultsCompared with the negative control group S, the integrated optical density (IOD) values of cranial dyed area in all groups increased significantly (P < 0.05); Compared with group S, the IOD value of cranial dyed area in mixtures of epimedium monomers increased significantly (P < 0.05). The mixtures of epimedium monomers were all effective in facilitating zebrafish cranial mineralization and preventing prednisolone from inducing osteoporosis. According to mixtures of A-K groups, zebrafish cranial mineralization gradually decreased with gradually reduced content of epimedin C, with significant difference among groups (P < 0.05). ConclusionThe higher the content of epimedin C in mixtures with invariant molarity is, the more active the anti-osteoporosis effect of epimedinC to zebrafish osteoporosis model is.