邻苯二甲酸二(2-乙基己基)酯暴露对斑马鱼胚胎的发育毒性及作用机制  

Developmental toxicity and molecular mechanism of Di-(2-ethylhexyl)phthalate exposure on zebrafish embryos

作　　者：钱显 冯伟伟 茆广华 陈瑶 罗梦娜 吴朝琼 仰榴青[2] 吴向阳 QIAN Xian;FENG Weiwei;MAO Guanghua;CHEN Yao;LUO Mengna;WU Chaoqiong;YANG Liuqing;WU Xiangyang(School of Environment and Safety Engineering,Jiangsu University,Zhenjiang Jiangsu 212013,China;School of Chemistry and Chemical Engineering,Jiangsu University,Zhenjiang Jiangsu 212013,China)

机构地区：[1]江苏大学环境与安全工程学院 [2]江苏大学化学化工学院,江苏镇江212013

出　　处：《江苏大学学报（医学版）》2024年第6期507-513,521,共8页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金面上项目(21976072)。

摘　　要：目的:研究邻苯二甲酸二(2-乙基己基)酯[di-(2-ethylhexyl)phthalate,DEHP]对斑马鱼胚胎发育的影响及潜在的分子机制。方法:将斑马鱼胚胎随机分为5组,分别为空白对照组、二甲基亚砜组以及8、40、200μg/L DEHP组,暴露至96 hpf,记录死亡率、孵化率、畸形率、心率、体长和胚胎自主运动次数等发育指标;采用酶学分析检测仔鱼体内超氧化物歧化酶(SOD)活力、谷胱甘肽过氧化氢酶(GPx)活力及丙二醛含量;采用实时荧光定量PCR法测定细胞凋亡、葡萄糖转运以及蛋白质、脂肪酸和糖原合成相关mRNA表达。结果:(1)与对照组相比,8μg/L DEHP组斑马鱼胚胎/仔鱼死亡率显著增加(P<0.01),仔鱼体长显著降低(P<0.05);40、200μg/L DEHP组斑马鱼胚胎/仔鱼死亡率和畸形率显著增加(P<0.01),孵化率、心率、体长以及胚胎自主运动次数明显降低(P<0.05或P<0.01)。(2)与对照组相比,8μg/L DEHP组斑马鱼仔鱼体内GPx活力显著降低(P<0.05);40、200μg/L DEHP组斑马鱼仔鱼体内SOD、GPx活力明显降低且丙二醛含量显著增加(P<0.01)。(3)与对照组相比,8μg/L DEHP组葡萄糖转运信号通路中Pi3k mRNA表达显著上调(P<0.01);40、200μg/L DEHP组凋亡信号通路中Bax/Bcl-2、Caspase-3和Ras mRNA表达显著上调(P<0.01),葡萄糖转运信号通路中Akt和Pi3k mRNA表达显著上调(P<0.01),Erk1/2 mRNA表达显著下调(P<0.05或P<0.01),蛋白质、脂肪酸和糖原合成信号通路中Mtor、Gsk-3α和Gsk-3βmRNA表达显著上调(P<0.05或P<0.01)。结论:DEHP可能通过诱导细胞凋亡,改变代谢相关mRNA表达,造成氧化损伤,从而影响斑马鱼胚胎的生长发育,产生发育毒性。Objective:To study the effects of di-(2-ethylhexyl)phthalate(DEHP)on embryonic development of zebrafish and its potential molecular mechanism.Methods:Zebrafish embryos were randomly divided into 5 groups:blank control group,dimethyl sulfoxide group,and 8,40,200μg/L DEHP groups,which were exposed to 96 hours post-fertilization(hpf).The mortality rate,hatching rate,malformation rate,heart rate,body length and the number of spontaneous movements of embryos were recorded.The activity of superoxide dismutase(SOD),glutathione catalase(GPx)and malondialdehyde(MDA)content in larvae were measured by enzymatic analysis.Real-time quantitative PCR was used to determine the expression of mRNA related to cell apoptosis,glucose transport,and the synthesis of proteins,fatty acids,and glycogen.Results:(1)Compared with the control group,the embryo/larvae mortality of zebrafish in 8μg/L DEHP group was significantly increased(P<0.01),and the larvae body length was greatly decreased(P<0.05);in 40 and 200μg/L DEHP groups,the mortality and malformation rate of zebrafish embryo/larvae were significantly increased(P<0.01),and the hatching rate,heart rate,body length and the number of embryonic autonomous movement were greatly decreased(P<0.05 or P<0.01).(2)Compared with the control group,the activity of GPx in zebrafish larvae in 8μg/L DEHP group was significantly decreased(P<0.05);the activities of SOD and GPx in zebrafish larvae were significantly decreased and the content of MDA was greatly increased in 40 and 200μg/L DEHP groups(P<0.01).(3)Compared with the control group,the expression of Pi3k mRNA in glucose transport signaling pathway in 8μg/L DEHP group was significantly up-regulated(P<0.01);the mRNA expressions of Bax/Bcl-2,Caspase-3 and Ras were greatly up-regulated(P<0.01),and the mRNA expressions of Akt and Pi3k in glucose transport signaling pathway were significantly up-regulated(P<0.01),the mRNA expression of Erk1/2 was greatly down-regulated(P<0.05 or P<0.01),and the mRNA expressions of Mtor,Gsk-3 and GSK-3βin prote

关 键 词：邻苯二甲酸二(2-乙基己基)酯 斑马鱼胚胎 发育毒性 分子机制 

分 类 号：R994[医药卫生—毒理学]