邻苯二甲酸二乙基己酯对斑马鱼胚胎心脏发育的毒性  

Toxic effect of di（2-ethylhexyl）-phthalate on zebrafish heart development

作　　者：李萌萌[1] 余章斌[1] 韩树萍[1] 胡晓山[1] 朱金改[1] 朱莎莎[1] 朱春[1] 郭锡熔[1] 

机构地区：[1]南京医科大学附属南京市妇幼保健院儿科,210004

出　　处：《中华实用儿科临床杂志》2013年第1期48-51,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（30973213）;江苏省医学重点人才基金（RC2011021）

摘　　要：目的探讨环境污染物邻苯二甲酸二乙基己酯（DEHP）对斑马鱼胚胎心脏发育的毒性及其机制。方法采用水域染毒法，将受精后4h（4hpf）的斑马鱼胚胎分别暴露于质量浓度为1、2、4、8mg／L的DEHP作为4个质量浓度组，同时设79．2mg／L甲醇对照组、60mg／L的氯化钠溶液空白对照组。通过分析72hpf的孵化率，24、48、72、96、120hpf的死亡率和畸形率等指标以及胚胎整体原位杂交的方法来评价DEHP对斑马鱼胚胎心脏发育的毒性。结果与对照组比较，随着DEHP质量浓度的增加，斑马鱼胚胎72hpf的孵化率降低，24、48、72、96、120hpf的死亡率和畸形率增加。当DEHP质量浓度达到8mg／L时，斑马鱼72hpf的孵出率由对照组的98．33％下降到62．22％，并且120hpf的死亡率达40．00％、畸形率达27．30％。整体原位杂交结果显示DEHP处理组斑马鱼胚胎心脏的位置和心管环化均出现不同程度的异常。与对照组比较，DEHP处理组胚胎在24hpf心房肌球蛋白重链（amhc）、心室肌球蛋白重链（vmhc）、心肌肌浆球蛋白轻链（cmlc2）的表达位置均发生了变化；48hpf amhc的表达量明显下降，vmhc的表达位置发生了变化，cmlc2标志的整个心脏发育异常，未能完成心脏环化过程，仅形成1条心管。结论DEHP能影响斑马鱼胚胎发育，导致斑马鱼胚胎和幼鱼的畸形与死亡，而它的心脏毒性可能发生在心管的环化过程中。Objective To determine the toxicity of the di（2-ethylhexyl）-phthalate （DEHP） on early develop- ment of zebrafish（danio rerio） embryos and to explore its mechanism during this procedure. Methods The fertilized embryos of zebrafish were divided into 6 groups ：4 groups of them were exposed to different concentrations of DEHP（ 1, 2,4,8 mg/L） ,while 2 other groups were treated with 79.2 mg/L methanol and 60 mg/L sodium chloride solution respectively as controls. To investigate the interaction between DEHP and early zebrafish embryonic development, the rate of hatching, malformation and lethality of embryos were studied from different DEHP-exposed and control groups. And to gain insight into the mechanisms underlying DEHP function in early zebrafish cardiac development,in situ hybridization for early cardiac molecular markers were performed. Results By carefully collecting and studying the results of DEHP-treated and control embryos,the rates of malformation of embryos and lethality of larvae were increased in a dose-dependent way,coupled with decreased hatching rate of embryos at 72 hpf. When the concentration of DEHP reached 8 mg/L,the hatching rate of embryo at 72 hpf was reduced to 62.22% ,the malformation rate of embryo was increased to 27.30% , and the lethality rate of larvae at 120 hpf reached 40.00%. Expression of cardiac specific gene, indicated by using in situ hybridization, showed that heart position and heart tube looping of zebrafish embryos from DEHP-treated groups were strongly affected. Compared with control embryos ,the expression of atrial myosin heavy chain（amhc） ,ventricular myosin heavy chain （vmhc） and cardiac myosin light chains （cmlc2） had been changed in the DEHP-treated groups at 24 hpf and 48 hpf. Conclusions DEHP could impact the development of zebrafish heart, and further lead to the malformation of zebrafish embryo and death of larvae. And the toxic effect of DEHP on zebrafish heart development may occur in the progress of heart tube looping.

关 键 词：斑马鱼 邻苯二甲酸二乙基己酯 心脏发育 胚胎毒性 

分 类 号：R114[医药卫生—卫生毒理学]