双酚A诱导内质网应激致甲状腺功能损害的作用机制研究  

Effects of thyroid dysfunction induced by Bisphenol A through endoplasmic reticulum stress

作　　者：夏瑞瑞 张姣香 齐从政 杨光宇[1] 张岩 李文学[1] 李军涛[1] 张波 胡前胜[2] 朱伟[1,5] XIA Rui-rui;ZHANG Jiao-xiang;QI Cong-zheng;YANG Guang-yu;ZHANC Yan;LI Wen-xue;LI Jun-tao;ZHANG Bo;HU Qian-sheng;ZHU Wei(Department of Toxicology,Guangzhou Center for Disease Control and Prevention,Guangzhou Guangdong 510440,China;School of Public Health,Sun Yat-sen University,Guangzhou Guangdong 510080,China;Southern Medical University,Guangzhou Guangdong 510515,China;Shenzhen Nanshan Center for Chronic Disease Control,Shenzhen Guangdong 518064,China;Institute of public health of Guangzhou Medical University,Guangzhou Guangdong 510635,China)

机构地区：[1]广州市疾病预防控制中心毒理学检验部,广东广州510440 [2]中山大学公共卫生学院预防医学系,广东广州510080 [3]南方医科大学公共卫生学院,广东广州510515 [4]深圳市南山区慢性病防治院,广东深圳518064 [5]广州医科大学公共卫生研究院,广东广州510635

出　　处：《毒理学杂志》2022年第6期471-477,489,共8页Journal of Toxicology

基　　金：国家自然科学基金面上项目(81773403);国家自然科学基金应急管理项目(81741101);广州市医学重点学科建设项目(20212023012)。

摘　　要：目的观察双酚A(Bisphenol A,BPA)染毒对成年雌性大鼠甲状腺功能的影响及甲状腺内质网应激机制在该过程中的作用。方法选用40只SD雌性大鼠,随机分为4组,分别经口灌胃给予玉米油(溶剂对照)和BPA,剂量分别为0.05、0.5和5 mg/kg·bw。10周时,麻醉处死动物并留取样品进行检测。采用实时荧光定量PCR法检测TG mRNA表达水平,采用蛋白质印迹法检测甲状腺球蛋白(thyroglobulin,TG)和内质网应激相关蛋白的表达水平的变化,采用苏木素-伊红(HE)染色进行甲状腺组织学检测及使用透射电子显微镜观察甲状腺滤泡细胞内的内质网结构的变化。结果BPA染毒可使实验动物血浆T3、T4和TG水平明显升高,低和中剂量染毒可明显上调甲状腺组织TG mRNA转录水平(P_(T3)=0.004,P_(T4)<0.001,PTG<0.001,PTG mRNA=0.001),并可致T3/T4比值明显下降,差异均有统计学意义(P<0.001)。Western blot检测结果显示BPA染毒可明显上调内质网应激中GRP78、ATF6和IRE1α蛋白水平(P_(GRP78)=0.008,P_(ATF6)=0.016,PIRE1α=0.001)。结论BPA染毒可诱导甲状腺功能紊乱,其中ATF6和IRE1通路激活及内质网应激可能是其机制之一。Objective To observe the effect of Bisphenol A exposure on thyroid function in adult female rats and the role of endoplasmic reticulum stress mechanism in this process.Methods Forty female SD rats were randomly divided into four groups according to body weight,with 10 rats in each group,and were exposed to BPA(0,0.05,0.5 and 5 mg/kg·bw,respectively)by oral gavage.At 10 weeks,the animals were anesthetized and executed to take samples for testing.The content of TG mRNA was detected by real-time fluorescence quantitative PCR and the expression levels of TG protein and endoplasmic reticulum stress related proteins were detected by Western blot.HE staining was used to detect the injury degree of thyroid tissue and transmission electron microscopy was used to observe the changes of endoplasmic reticulum structure in thyroid follicular cells.Results The levels of T3,T4 and TG in plasma.Low dose and medium dose exposure can significantly up-regulate TG mRNA transcription levels in thyroid tissue(P_(T3)=0.004,P_(T4)<0.001,PTG<0.001,PTG mRNA=0.001),and significantly decreased the T3/T4 ratio(P<0.001),but had no significant effect on plasma TSH and TG protein in tissues.The levels of GRP78,ATF6 and IRE1αin ER stress were significantly up-regulated by exposure to Bisphenol A(P_(GRP78)=0.008,P_(ATF6)=0.016,P_(IRE1α)=0.001),but,p-eIF2αand Caspase-12 and Caspase-9 apoptotic proteins were not significantly affected.Conclusion Bisphenol A exposure can induce thyroid dysfunction and activate ATF6 and IRE1 pathways to induce endoplasmic reticulum stress.

关 键 词：甲状腺激素 甲状腺球蛋白 双酚A 内质网应激 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]