生命早期6:2 Cl-PFESA暴露对子代小鼠海马AMPA受体基因表达的影响  

Effects of early-life 6:2 Cl-PFESA exposure on hippocampal AMPA receptor gene expression in offspring mice

作　　者：王瑞文 姚雅平 蔡景双 曲芙林 金晓霞 WANG Ruiwen;YAO Yaping;CAI Jingshuang;QU Fulin;JIN Xiaoxia(Department of Occupational Health and Occupational Medicine,School of Public Health,Shenyang Medical College,Shenyang,Liaoning 110034,China)

机构地区：[1]沈阳医学院公共卫生学院职业卫生与职业医学教研室,辽宁沈阳110034

出　　处：《环境与职业医学》2024年第5期552-559,共8页Journal of Environmental and Occupational Medicine

基　　金：辽宁省自然科学基金项目(2021-BS-285);沈阳医学院大学生科研课题(20239066)。

摘　　要：[背景]6:2氯代多氟醚基磺酸(6:2 Cl-PFESA)具有十分强大的生物蓄积性和胎盘屏障穿透力,还可穿过血脑屏障。然而,其生命早期暴露对子代产生神经发育毒性的机制尚为未知。[目的]通过建立6:2 Cl-PFESA暴露动物模型,探究生命早期6:2 Cl-PFESA暴露对仔鼠生长发育及海马α-氨基-3-羟基-5-甲基-4-异恶唑丙酸(AMPA)受体基因表达的影响。[方法]将30只昆明种孕鼠随机分为对照组以及2、10、50和250μg·L^(-1)6:2 Cl-PFESA暴露组,从受孕第1天开始以自由饮水方式暴露相应剂量的6:2 Cl-PFESA,至泌乳结束。仔鼠于出生后(PND)第21天断乳,通过自由饮水方式继续进行6:2 Cl-PFESA暴露。记录仔鼠出生体重和体长作为本次实验基础数据。各组仔鼠于PND7、PND21和PND35分别麻醉后处死,取脑组织并剥离海马,透射电镜观察神经元突触超微结构,实时荧光逆转录聚合酶链式反应法检测AMPA受体GluR1、GluR2和GluR3的基因表达变化情况。PND35仔鼠于处死前进行Morris水迷宫实验观察仔鼠空间学习记忆能力情况。[结果]10、50、250μg·L^(-1)6:2 Cl-PFESA暴露组仔鼠的出生体重和体长分别为(2.23±0.36)、(1.92±0.20)、(1.88±0.31)g和(33.73±0.98)、(32.91±1.30)、(32.52±2.07)mm,均低于对照组(2.78±0.35)g和(36.46±2.34)mm(P<0.05)。PND35仔鼠Morris水迷宫结果显示:定位航行实验第4天的250μg·L^(-1)6:2 Cl-PFESA暴露组、第5天的10、50以及250μg·L^(-1)6:2 Cl-PFESA暴露组仔鼠较比对照组逃避潜伏期延长(P<0.05);空间探索实验中,50和250μg·L^(-1)6:2 ClPFESA暴露组仔鼠的穿越平台次数、250μg·L^(-1)6:2 Cl-PFESA暴露组仔鼠的目标象限停留时间与对照组相比均减少(P<0.05)。经透射电镜观察发现:PND35的250μg·L^(-1)6:2 Cl-PFESA暴露组与对照组相比,仔鼠突触后致密物变薄、突触间隙增宽。不同发育阶段的250μg·L^(-1)6:2Cl-PFESA暴露组仔鼠海马中GluR1、GluR2和GluR3的mRNA表达水平与对照组相[Background]The compound 6:2 chlorinated polyfluorinated ether sulfonic acids(6:2 Cl-PFESA)has been demonstrated abilities of strong bioaccumulation and placental barrier penetration,and it can also cross the blood-brain barrier.However,the mechanism of its neurodevelopmental toxicity in offspring induced by early-life exposure is still unknown.[Objective]To explore effects of 6:2 Cl-PFESA on the growth and theα-amino-3-hydroxy-5-methylisoxazole-4-propionic(AMPA)receptor gene expression in the hippocampus of offspring mice by establishing a 6:2 Cl-PFESA exposure animal model.[Methods]Thirty Kunming pregnant mice were randomly divided into five groups:control group,and 2,10,50,and 250μg·L^(-1)6:2 Cl-PFESA exposure groups.The treatment groups were exposed to designed doses of 6:2 Cl-PFESA through drinking water from the first day of gestation until the end of lactation.The pups were weaned on postnatal day(PND)21,and continued to be exposed to 6:2 Cl-PFESA through drinking water.Birth weight and body length of the offspring were recorded.Offspring mice were anesthetized and sacrificed re-spectively on PND7,PND21,and PND35,then their hippocampus was peeled from harvested brain tissue.The ultrastructure of hippocampus was observed via transmission electron microscopy;and the expression of AMPA receptors GluR1,GluR2,and GluR3 in the hippocampus was evaluated by real-time reverse transcription polymerase chain reaction.The learning and memory ability of the PND35 mice was measured by Morris water maze test before they were sacrificed.[Results]The birth weights and the lengths of the pups in the 10,50,and 250μg·L^(-1)6:2 Cl-PFESA exposure groups were(2.23±0.36),(1.92±0.20),(1.88±0.31)g,and(33.73±0.98),(32.91±1.30),(32.52±2.07)mm,respectively,which were lower than those in the control group,(2.78±0.35)g and(36.46±2.34)mm(P<0.05),respectively.The results of Morris water maze showed that the escape latencies in the orientation navigation experiment on the 4th day in the 250μg·L^(-1)6:2 Cl-PFESA exposure grou

关 键 词：α-氨基-3-羟基-5-甲基-4-异恶唑丙酸受体 6:2氯代多氟醚基磺酸 神经发育毒性 突触可塑性 学习记忆 

分 类 号：R114[医药卫生—卫生毒理学]