[Activity Study of Icarlin and Epimedin C Monomer under the Same Molarity in Osteoporosis Ze- brafish Model].

Objective To evaluate anti-osteoporotic activity of icariin and Epimedin C monomer under the same molarity in predinsolone-induced osteoporosis zebrafish. Methods Zebrafish larvae after 4-day fertilization were divided into group S [0. 5% dimethyl sulfoxide (DMSO) , A (25 µmol/L prednisolone, 0. 5% DMSO), B (2 IU/L salmon calcitonin, 25 µmol/L prednisolone,0. 5% DMSO), C (1. 5 1,mol/L icariin, 25 µmol/L prednisolone, 0. 5% DMSO) , D (15 µLmol/L icariin,25 µmol/L prednisolone, 0. 5% DM- SO), E (150 µmol/L icariin, 25 µmol/L prednisolone, 0. 5% DMSO), F (1. 5 µmol/L Epimediri C, 25 µmol/L prednisolone, 0. 5% DMSO) , G (15 µmol/L Epimedin C, 25 µmol/L prednisolone, 0.5% DM- SO) , H (150 µmol/L Epimedin C, 25 µmol/L prednisolone, 0. 5% DMSO). All culture solution contained 0. 5% DMSO. All the young fishes were grown in a 24-well plate. The culture medium was changed every day. They were cultured in a incubator box at 28. 5 °C and killed at day 9. Zebrafish skeleton was stained with alizarin red. The stained Zebrafish ventral skull was observed using microscope, and mineralized area was quantitatively analyzed. Results Compared with group S, accumulative integrated optical densi- ty(IOD)of the mineralized area significantly decreased in group A (P <0. 01) ; accumulative IOD of the mineralized area significantly increased in group B (P <0. 01). The accumulative IOD of the mineralized area showed weakly increasing tendency in group C, D, and E along with increased concentration (P < 0. 05). Compared with group A, accumulative IOD obviously increased in group B with statistical difference (P <0. 01) , but with no statistical difference as compared with group C or group D (P >0. 05). Statistical difference existed in accumulative IOD between group A and group E (P <0. 05). The mineralized area showed increasing tendency in group F and group G along with increased concentration (P <0. 05), and accumulative IOD obviously increased as well (P <0. 05). No Zebrafish embryo survived in group H. There was no statistical difference in Zebrafish embryo survival among group E, F, or G (P >0. 05). The staining of Zebrafish skull was clearly seen in group S, with vertebrae and bilateral branchial skeleton clearly seen. The intensity of staining in the same area was obviously attenuated in group A. The osteo- genesis was speeded up under the same condition in group B, with obviously enlarged mineralized area and more darkly stained bone tissue. The mineralization of skull was gradually increasing during the stai- ning process in group C, D, E, F, and G. The mineralized area and the intensity of staining were gradually enhanced, and changes of vertebrae were most obviously seen in group C, D, E, F, and G, but they were not arrived at the stained intensity level in group B. Conclusions Osteoporosis Zebrafish model is a simple and efficient model for screening bioactive ingredients of Chinese herbs. The activity of Epimedin C at low concentration was better than icariin in this model. But possible toxicity of Epimedin C at high concentration needs to be further studied.

Androgen signaling inhibits de novo lipogenesis to alleviate lipid deposition in zebrafish.

Testosterone is closely associated with lipid metabolism and known to affect body fat composition and muscle mass in males. However, the mechanisms by which testosterone acts on lipid metabolism are not yet fully understood, especially in teleosts. In this study, cyp17a1-/- zebrafish ( Danio rerio) exhibited excessive visceral adipose tissue (VAT), lipid content, and up-regulated expression and activity of hepatic de novo lipogenesis (DNL) enzymes. The assay for transposase accessible chromatin with sequencing (ATAC-seq) results demonstrated that chromatin accessibility of DNL genes was increased in cyp17a1-/- fish compared to cyp17a1+/+ male fish, including stearoyl-CoA desaturase ( scd) and fatty acid synthase ( fasn). Androgen response element (ARE) motifs in the androgen signaling pathway were significantly enriched in cyp17a1+/+ male fish but not in cyp17a1-/- fish. Both androgen receptor ( ar)-/- and wild-type (WT) zebrafish administered with Ar antagonist flutamide displayed excessive visceral adipose tissue, lipid content, and up-regulated expression and activity of hepatic de novo lipogenesis enzymes. The Ar agonist BMS-564929 reduced the content of VAT and lipid content, and down-regulated acetyl-CoA carboxylase a ( acaca), fasn, and scd expression. Mechanistically, the rescue effect of testosterone on cyp17a1-/- fish in terms of phenotypes was abolished when ar was additionally depleted. Collectively, these findings reveal that testosterone inhibits lipid deposition by down-regulating DNL genes via Ar in zebrafish, thus expanding our understanding of the relationship between testosterone and lipid metabolism in teleosts.