诺氟沙星对斑马鱼胚胎发育的毒性作用及对TGF-β1的影响

The Deleterious Effects of Norfloxacin on Zebrafish Embryonic Development and TGF-β1 Expression

旨在通过观察不同浓度诺氟沙星对斑马鱼胚胎不同发育时期的毒性作用,以及对TGF-β1基因表达的影响。配制诺氟沙星浓度为0、10、20、40μmol/L,将斑马鱼胚胎暴露在上述浓度的诺氟沙星中。观察在胚胎不同发育时期,诺氟沙星对斑马鱼脊柱弯曲、心包囊肿、卵黄囊肿和死亡率的影响,以及使用实时定量聚合酶链式反应(q PCR)检测其对TGF-β1基因表达的影响。结果显示,诺氟沙星对斑马鱼的胚胎发育有明显影响,主要表现在脊柱弯曲、心包囊肿和卵黄囊肿,随着诺氟沙星暴露浓度的增大,胚胎的发育延迟,孵化时间延长,胚胎死亡率增加;当诺氟沙星暴露浓度为40μmol/L时,96 hpf的胚胎死亡率达到76.45%;与正常状态相比,暴露于不同浓度诺氟沙星的斑马鱼胚胎中TGF-β1基因的m RNA表达随发育时间延长而增加趋势减缓。说明诺氟沙星对斑马鱼胚胎发育的致畸作用与致死作用有显著的影响。提示水体中残留的诺氟沙星对鱼类的生殖与发育具有潜在的危害。

The aim of this work is to investigate the deleterious effects of norfloxacin on zebrafish embryonic development in different periods, and determine its effects on TGF-β1 expression. Zebrafish embryos were exposed to different concentrations of norfloxacin（0, 10, 20, and 40 μmol/L）. The effects of norfloxacin on spinal curvature, pericardial cyst, yolk edema and mortality of zebrafishes in the different developmental periods were observed. Furthermore, the m RNA level of TGF-β1 was examined by q PCR. The results showed that norfloxacin had significantly deleterious effects on zebrafish embryonic development, as mainly evidenced by spinal curvature, pericardial cyst and yolk edema. With the increase of concentration of norfloxacin, embryonic development was delayed, and the hatch period and mortality increased. The mortality rate of zebrafish embryos at 96 hpf re ached 76.46% after exposure to 40 μmol/L norfloxacin. Moreover, comparing with control group, the increase trend on m RNA level of TGF-β1 in zebrafish embryo exposing to different dose of norfloxacin slowed down with the prolonging of development time. Conclusively, norfloxacin has teratogenic and lethal effects on zebrafish embryonic development, suggesting norfloxacin residue in water system has serious latent dangers to the reproduction and development of fish.