纳米氧化铈对斑马鱼胚胎早期发育毒性

Toxic effect of nano-cerium oxide on zebrafish embryonic early development

目的探讨纳米氧化铈对斑马鱼胚胎早期发育的影响及机制。方法将生长发育良好的斑马鱼胚胎随机分为对照组和纳米氧化铈染毒50、100、200、400、800 mg/L剂量组,每组40个。各剂量组斑马鱼胚胎分别予相应质量浓度的纳米氧化铈染毒5 d,对照组斑马鱼胚胎不予染毒。观察斑马鱼胚胎死亡以及畸形情况,采用激光共聚焦显微镜实时记录斑马鱼胚胎心率,采用蛋白免疫印迹实验检测斑马鱼胚胎活化的含半胱氨酸天冬氨酸蛋白水解酶3(cleaved Caspase-3)、微管相关蛋白1轻链3(LC3)蛋白的表达情况。结果各组斑马鱼胚胎死亡率、胚胎畸形率随染毒剂量增大而增加(P<0.01);800 mg/L剂量组斑马鱼胚胎心率低对照组[(77±8)vs(93±4)次/分,P<0.01]。各组斑马鱼胚胎LC3-Ⅱ蛋白相对表达水平比较,差异无统计学意义(P>0.05)。各剂量组斑马鱼胚胎cleaved Caspase-3蛋白相对表达水平均高于对照组(P<0.05);200 mg/L剂量组斑马鱼胚胎cleaved Caspase-3蛋白相对表达水平高于50 mg/L剂量组(P<0.05)。结论纳米氧化铈颗粒可能通过激活细胞凋亡,引起斑马鱼胚胎早期发育的毒性效应。

Objective To study the effect of nano-cerium oxide on the early development of zebrafish embryos.Methods The well-developed zebrafish embryos were randomly divided into the control group and the 50,100,200,400 and 800 mg/L dose groups,with 40 embryos in each group.The dose groups were treated with nano-cerium oxide at the corresponding mass concentration for 5 days.The control group received no treatment.The death and malformation of embryos were observed.The heart rate of zebrafish embryos was recorded using confocol microscope.The protein expression of microtubule-associated protein 1 light chain 3(LC3)and cleaved Caspase-3 and were observed by Western blot technology.Results The death and embryonic malformation rate of zebrafish embryos increased with the increase of doses,showing statistical significance(P<0.01).The heart rate of the 800 mg/L dose group was decreased compared with the control group[(77±8)vs(93±4)beats/min,P<0.01].There was no statistical significant difference in LC3-Ⅱprotein expression in each groups(P>0.05).The cleaved Caspase-3 protein expression increased in all dose groups compared with the control group(P<0.05).The cleaved Caspase-3 protein expression in the 200 mg/L dose group was higher than that in the 50 mg/L dose group(P<0.05).Conclusion The nano-cerium oxide may induce cell apoptosis,causing toxic effect in early development of zebrafish embryos.