磷酸三苯酯及磷酸甲苯二苯酯的早期胚胎发育毒性:PPARγ的作用  

Early embryonic developmental toxicity of TPhP and CDP:Roles of PPARγ

作　　者：黄静[1] 翟雨诺 李博洋 纪静[1] 李传海 刘世昕 刘议蔓 袁骏华 姜启晓 HUANG Jing;ZHAI Yunuo;LI Boyang;JI Jing;LI Chuanhai;LIU Shixin;LIU Yiman;YUAN Junhua;JIANG Qixiao(School of Public Health,Qingdao University,Qingdao,Shandong 266071,China;School of Basic Medicine,Qingdao University,Qingdao,Shandong 266071,China)

机构地区：[1]青岛大学公共卫生学院,山东青岛266071 [2]青岛大学基础医学院,山东青岛266071

出　　处：《环境与职业医学》2024年第12期1369-1375,1383,共8页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金项目(82100854);山东省博士后创新项目(SDCX-ZG-202400024)。

摘　　要：[背景]有机磷阻燃剂属于新兴环境污染物,已有多种毒性报道,但其对胚胎发育毒性的研究较少。亟待阐明相关效应及机制,以期完善风险评估,更好地保护敏感人群。[目的]评价磷酸三苯酯(TPhP)和磷酸甲苯二苯酯(CDP)暴露对早期鸡胚的潜在发育毒性,以鸡胚报告基因表达系统揭示TPhP和CDP在体激活过氧化物酶体增殖物激活受体γ(PPARγ)的能力,并以慢病毒在体沉默为工具,探讨PPARγ在TPhP和CDP诱导鸡胚发育毒性中的作用。[方法]首先采用不同剂量的TPhP及CDP对受精鸡蛋进行气室注射染毒,孵化6d后评价不同剂量下鸡胚发育情况,选取最佳剂量进行后续实验;接着采用报告基因系统来评价TPhP及CDP对PPARγ的胚胎内激活情况;最后采用慢病毒沉默PPARγ,与TPhP及CDP染毒共处理,进一步揭示PPARγ在观察到的发育毒性中所发挥的作用。[结果]TPhP及CDP发育染毒后,在孵化6 d时观察到10、30 mg·kg^(-1)的TPhP及3、10、30 mg·kg^(-1)的CDP染毒引起鸡胚重量(以蛋重标准化)降低(P<0.05),提示二者均具有一般发育毒性,CDP毒性较高。此外CDP染毒还引起鸡胚矢状面脑区面积(以鸡胚体重标准化)增加(P<0.05),而鸡胚矢状面眼区面积(以鸡胚体重标准化)减小(P<0.05),提示CDP具有特异性的神经及眼发育毒性。报告基因实验结果显示,阳性对照罗格列酮、TPhP、CDP与对照相比均能激活PPARγ(P<0.05),效应强度大小:罗格列酮>CDP>TPhP。以PPARγ沉默慢病毒实现鸡胚PPARγ在体沉默,实时荧光定量多聚核苷酸链式反应(qRT-PCR)结果显示沉默效率约为55%。PPARγ在体沉默可以有效地缓解TPhP及CDP染毒引起的体重下降(P<0.05)以及CDP染毒引起的脑区扩大及眼球发育不良(P<0.05)。[结论]TPhP及CDP染毒在早期鸡胚均可引起一般发育毒性,CDP毒性较高。同时,CDP还引起特异性的脑区增大及眼球发育不良,其机制与PPARγ的激活有关。[Background]Organic phosphate flame retardants are emerging environmental pollutants.While there have been multiple toxicities reported following organic phosphate flame retardants exposure,few studies focus on their potential developmental toxicities.It is necessary to elucidate these developmental toxicological effects and underlying mechanisms to improve risk assessments and better protect sensitive populations.[Objective]To evaluate potential developmental toxicities in early chicken embryos following exposure to triphenyl phosphate(TPhP)or cresyl diphenyl phosphate(CDP),to reveal TPhP and CDP’s capabilities to activate peroxisome proliferator-activated receptorγ(PPARγ)in vivo in an established chicken embryo gene reporter system,and to investigate the roles of PPARγin TPhP/CDP-induced developmental toxicities with lentivirus-mediated in vivo gene silencing.[Methods]Firstly,diverse doses of TPhP and CDP were injected into the air sacs of fertilized eggs to assess the development of chicken embryos after 6 d of incubation,and an optimal dose was chosen for subsequent experiments.Subsequently,the report gene system was employed to evaluate the intraembryonic activation of PPARγby TPhP and CDP.Eventually,PPARγwas silenced using lentivirus,and the embryos were co-treated with TPhP and CDP to further disclose the roles of PPARγin the observed developmental toxicity.[Results]Following developmental exposure to TPhP or CDP,significantly lower chicken embryo weights(normalized with egg weights)were observed in the 6 d embryos(10,30 mg·kg^(-1) TPhP and 3,10,30 mg·kg^(-1) CDP),indicating that both chemicals have general developmental toxicities and CDP is more potent.Additionally,exposure to CDP also resulted in remarkably increased sagittal brain area(normalized to embryo weights)and decreased sagittal eye area(normalized to embryo weights)(P<0.05),suggesting that CDP has specific developmental neurotoxicity and ocular toxicity.The PPARγreporter gene experiment results revealed that rosiglitazone(positive c

关 键 词：磷酸三苯酯 磷酸甲苯二苯酯 发育毒性 鸡胚 过氧化物酶体增殖物激活受体Γ 

分 类 号：R11[医药卫生—公共卫生与预防医学]