论文部分内容阅读
目的用泼尼松龙诱导的斑马鱼骨质疏松模型评价相同摩尔浓度下淫羊藿苷和朝藿定C单体的抗斑马鱼骨质疏松作用。方法将受精后4日的斑马鱼胚胎分为S组(0.5%二甲亚砜DMSO)、A组(泼尼松龙25μmol/L,0.5%DMSO)、B组(2IU/L鲑降钙素,泼尼松龙25μmol/L,0.5%DMSO)、C组(淫羊藿苷1.5μmol/L,泼尼松龙25μmol/L,0.5%DMSO)、D组(淫羊藿苷15μmol/L,泼尼松龙25μmol/L,0.5%DMSO)、E组(淫羊藿苷150μmol/L,泼尼松龙25μmol/L,0.5%DMSO)、F组(朝藿定C 1.5μmol/L,泼尼松龙25μmol/L,0.5%DMSO)、G组(朝藿定C 15μmol/L,泼尼松龙25μmol/L,0.5%DMSO)、H组(朝藿定C 150μmol/L,泼尼松龙25μmol/L,0.5%DMSO)。所有培养液均含有0.5%DMSD。将各组幼鱼放置与24孔板中,每日更换培养液,在恒温环境28.5℃培养箱中,培养至第9天处死,以茜素红染色。用显微镜对斑马鱼颅骨腹侧进行观察,定量分析将成像染色区域。结果与S组比较,A组骨矿化累计光密度值降低(P<0.01);B组骨矿化累计光密度值升高(P<0.01)。而C、D、E组随浓度增加矿化面积呈微弱的递增趋势(P<0.05)。与A组比较,B组累计光密度值增高明显,差异有统计学意义(P<0.01),C、D组差异无统计学意义(P>0.05),而E组明显升高(P<0.05)。F、G组随浓度增加,矿化面积呈递增趋势,颅骨染色累计光密度值升高(P<0.05),H组内斑马鱼胚胎无存活。E、F、G组间比较,差异无统计学意义(P>0.05)。S组斑马鱼头颅骨染色清晰,脊椎骨及两旁鳃骨染色清晰。A组在相同染色区域的强度明显减少。B组在相同条件下呈现骨组织成骨加快,矿化面积明显增多,骨组织深染等特点。C、D、E、F、G组,在染色中颅骨矿化程度逐渐增加,椎骨及两侧腮骨矿化面积及染色强度递增,以椎骨改变最为显著,但均未达到B组的染色强度。结论斑马鱼骨质疏松模型是一种简单、高效的中药成分筛选模型,低浓度朝藿定C在该模型中的活性优于淫羊藿苷,高浓度朝藿定C的可能毒性作用还需要深入研究。
OBJECTIVE: To evaluate the anti-zebrafish osteoporosis effect of icariin and epimedin C at the same molar concentration using prednisolone-induced zebrafish osteoporosis model. Methods Four days after fertilization, zebrafish embryos were divided into group S (DMSO 0.5%), group A (25μmol / L prednisolone 0.5% DMSO), group B (2IU / L salmon calcitonin , 25μmol / L prednisolone (25μmol / L, 0.5% DMSO), C group (15μmol / L icariin, Prednisolone 25μmol / L, 0.5% DMSO), group E (icariin 150μmol / L, prednisolone 25μmol / (30 μmol / L prednisolone, 25 μmol / L prednisolone and 0.5% DMSO) in group G (epimedin C 150 μmol / L, prednisolone 25 μmol / Dragon 25 μmol / L, 0.5% DMSO). All broths contained 0.5% DMSD. The larvae of each group were placed in a 24-well plate, and the medium was changed daily. In a constant temperature environment of 28.5 ° C incubator, the cells were cultured until the 9th day, and were stained with alizarin red. The zebrafish skull was ventrally observed with a microscope and the area of imaging staining was quantified. Results Compared with group S, the cumulative optical density of bone mineralization in group A decreased (P <0.01), and that of group B increased (P <0.01). While the mineralized area of C, D and E groups showed a weak increasing trend with increasing concentration (P <0.05). Compared with group A, the cumulative optical density of group B increased significantly (P <0.01), but there was no significant difference between group C and D (P> 0.05), while group E increased significantly (P <0.05) ). F and G groups increased with the increase of concentration, the area of mineralization showed an increasing trend, the accumulated optical density of skull increased (P <0.05), and the zebrafish embryos of group H did not survive. There was no significant difference between E, F and G groups (P> 0.05). S group zebrafish skull staining, vertebrae and both sides of the gill bone staining clear. The intensity of group A in the same staining area decreased significantly. Group B under the same conditions, bone tissue showed accelerated bone mineralization increased significantly, such as bone tissue stained and so on. C, D, E, F and G groups. Skull mineralization gradually increased in staining. The mineralization area and staining intensity of vertebrae and both sides of the cheekbone gradually increased. The change of vertebrae was the most significant, but the staining intensity of group B did not reach . Conclusion Osteoporosis model of zebrafish is a simple and effective screening model of traditional Chinese medicine ingredients. The low concentration of epimedin C in this model is superior to icariin, and the possible toxicological effects of high concentration of epimedin C In-depth study.