芳香烃受体介导四溴双酚A致斑马鱼成鱼心脏毒性作用  

Aryl Hydrocarbon Receptor-mediated Cardiac Toxicity in Adult Zebrafish Induced by Tetrabromobisphenol A

作　　者：单婧 马小发 吴梦宇 林雨佳 王义 王蕊 徐海明 SHAN Jing;MA Xiaofa;WU Mengyu;LIN Yujia;WANG Yi;WANG Rui;XU Haiming(School of Public Health and Management,Ningxia Medical University,Yinchuan 750004,China;The Key Laboratory of Environmental Factors and Chronic Disease Control of Ningxia,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院,银川750004 [2]宁夏回族自治区环境因素与慢性病控制重点实验室,银川750004

出　　处：《宁夏医科大学学报》2023年第4期325-331,共7页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(81660527);宁夏回族自治区重点研发计划项目(2021BEG02030);2020年宁夏青年科技人才托举工程;2020年宁夏医科大学青年骨干人才培育计划。

摘　　要：目的以模式生物斑马鱼作为实验动物,使用芳香烃受体(AhR)抑制剂CH223191(CH),探究AhR在四溴双酚A(TBBPA)致斑马鱼心脏毒性效应中的作用。方法将斑马鱼成鱼随机分为溶剂对照组(DMSO组)、CH组(0.05μmol·L^(-1))、TBBPA暴露组(10、100、1000 nmol·L^(-1))和CH+TBBPA干预组(1000 nmol·L^(-1)),暴露持续30 d。暴露结束后,称重并测量体长,采用qRT-PCR方法检测心脏组织中AhR相关基因(ahr2、cyp1a1和cyp1a2)以及心脏功能关键基因(sox9b、nkx2.5、gate4、myh6、tbx5和vmhc)以及氧化应激相关基因(nrf2、gpx1a、sod1、sod2和cat)的表达水平,使用试剂盒检测肝组织中抗氧化酶(SOD、CAT和GSH-Px/GPX)活性。结果与DMSO对照组相比,各暴露组体质量、体长差异均无统计学意义(P均>0.05)。AhR相关基因ahr2、cyp1a1和cyp1a2在CH组中表达下调,在10、100、1000 nmol·L^(-1) TBBPA组表达均上调(P均<0.05)。与1000 nmol·L^(-1) TBBPA组相比,上述AhR相关基因在CH+1000 nmol·L^(-1) TBBPA组中表达水平均下降(P均<0.05)。在1000 nmol·L^(-1) TBBPA组中,心脏功能关键基因sox9b、nkx2.5、gate4、myh6、tbx5和vmhc表达均下调(P均<0.05),氧化应激相关基因gpx1a、sod1、sod2和cat表达均下调,nrf2表达上调(P均<0.05)。抗氧化酶活性检测结果显示,1000 nmol·L^(-1) TBBPA组斑马鱼肝组织中SOD、CAT和GSH-Px/GPX活性均下降(P均<0.05),分别相当于DMSO对照组的0.45、0.57和0.44倍。与1000 nmol·L^(-1) TBBPA组相比,CH+1000 nmol·L^(-1) TBBPA组各抗氧化酶活性均有所上升(P均<0.05)。结论环境相关剂量的TBBPA可能通过激活AhR信号通路,引起氧化损伤并影响心脏功能关键基因的表达水平,最终导致心脏毒性。Objective In this study,zebrafish(Danio rerio),a model organism,was used as an experimental animal,and CH223191(CH),an aromatic hydrocarbon receptor(AhR)inhibitor,was used to explore the role of AhR in the cardiac toxic effect of tetrabromobisphenol A(TBBPA)on zebrafish.Methods Adult zebrafish were randomly divided into six groups for exposure,namely solvent control group(DMSO),CH group(0.05μmol·L^(-1)),TBBPA exposure groups(10,100,1000 nmol·L^(-1)),and CH+TBBPA(1000 nmol·L^(-1))group.After 30 days of exposure,the body weight and body length were measured.The expression of AhR-related genes(ahr2,cyp1a1 and cyp1a2),cardiac function-related genes(sox9b,nkx2.5,gate4,myh6,tbx5 and vmhc)and reactive oxygen species-related genes(nrf2,gpx1a,sod1,sod2 and cat)in cardiac tissue were detected by qRT-PCR.The antioxidant enzymes(SOD,CAT and GSH-Px/GPX)activities in liver tissue were tested by commercial kits.Results Compared with the DMSO control group,there was no significant difference in body weight and body length in all exposure groups(P all>0.05).The results of qRT-PCR showed that AhR-related genes(ahr2,cyp1a1 and cyp1a2)was down-regulated in the CH group,while up-regulated in 10,100,1000 nmol·L^(-1) TBBPA groups(P all<0.05).Compared with the 1000 nmol·L^(-1) TBBPA exposure group,these AhR related genes were decreased to a certain extent in the CH+1000 nmol·L^(-1) TBBPA group(P all<0.05).The cardiac function-related genes(sox9b,nkx2.5,gate4,myh6,tbx5 and vmhc)were down-regulated in the 1000 nmol·L^(-1) TBBPA group(P all<0.05).The reactive oxygen species-related genes(gpx1a,sod1,sod2 and cat)were down-regulated while nrf2 was up-regulated in the 1000 nmol·L^(-1) TBBPA group(P all<0.05).The results of the antioxidant enzyme activities showed that hepatic SOD,CAT and GSH-Px/GPX activities were decreased in the 1000 nmol·L^(-1) TBBPA group(P all<0.05).Specifically,the activities of these enzymes were equivalent to 0.45 times,0.57 times and 0.44 times of the DMSO control group respectively.Compared with the 1000 n

关 键 词：四溴双酚A 芳香烃受体 斑马鱼 心脏毒性 氧化应激 

分 类 号：R114[医药卫生—卫生毒理学]