Cu^(2+)、Pb^(2+)对青海湖裸鲤的急性毒性效应及相关基因表达的影响  

Toxicity effects of Cu^(2+)and Pb^(2+)on Gymnocypris przewalskii and regulation gene expression

作　　者：李梓瑄 金文杰 李长忠 LI Zi-xuan;JIN Wen-jie;LI Chang-zhong(College of Eco-Environmental Engineering,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学生态环境工程学院,西宁810016

出　　处：《淡水渔业》2023年第5期41-54,共14页Freshwater Fisheries

基　　金：国家自然科学基金地区项目(31460700);青海省重大科技专项(2019-NK-A2);青海大学生态环境工程学院实验教学创新项目(2018-STXY-B-b2)。

摘　　要：为了研究Cu^(2+)、Pb^(2+)对青海湖裸鲤(Gymnocypris przewalskii)的急性毒性效应及相关基因相对表达水平的变化,实验以静态暴露和qPCR法分别进行毒性试验及检测青海湖裸鲤鳃、肾脏和肝脏中Nrf2(核因子NF-E2相关因子)、Keap1(Kelch样环氧氟丙烷相关蛋白)、MT(金属硫蛋白)、TNF-α(肿瘤坏死因子α)、Hsp90(热休克蛋白90)、Hsp70(热休克蛋白70)、iNOS(诱导型一氧化氮合酶)和COX-2(环氧化酶2)的相对表达。毒性实验结果表明,Cu^(2+)(0.1、0.2、0.5、1和2 mg/L)对1龄青海湖裸鲤安全浓度为0.03 mg/L;Pb^(2+)(0.5、1、2.5、5和10 mg/L)安全浓度为0.1 mg/L;Cu^(2+)+Pb^(2+)的复合胁迫对青海湖裸鲤具有拮抗作用。在低于安全浓度Cu^(2+)、Pb^(2+)与Cu^(2+)+Pb^(2+)复合胁迫下,检测基因表达水平结果显示,Nrf2在Cu^(2+)和Pb^(2+)胁迫12 h时表达上调,Keap1在Cu^(2+)、Pb^(2+)与Cu^(2+)+Pb^(2+)复合胁迫12 h的肾脏中显著下调,表明Nrf2-Keap1两者之间存在负调控作用。各浓度条件下,MT的表达在鳃和肝脏中均下调,TNF-α的表达在鳃中上调。在Cu^(2+)+Pb^(2+)复合条件下,TNF-α的表达显著上调。在Cu^(2+)、Pb^(2+)条件下,Hsp90、Hsp70、iNOS和COX-2的表达量均下调。结果表明,各浓度Cu^(2+)、Pb^(2+)与Cu^(2+)+Pb^(2+)对青海湖裸鲤均产生毒性影响,各基因均对低浓度重金属离子作用产生应答。The relative expression level of related genes,static and qPCR methods was detected to reveal the relative expression of Nrf2,Keap1,MT,TNF-α,Hsp90,Hsp70,iNOS and COX-2 in gill,kidney and liver of Gymnocypris przewalskii to study the acute toxicity effect of Cu ^(2+)and Pb ^(2+)on G.przewalskii.The toxicity test showed that the safe concentration of Cu ^(2+)(0.1,0.2,0.5,1 and 2 mg/L)in the first instar G.przewalskii was 0.03 mg/L.The safe concentration of Pb ^(2+)(0.5,1,2.5,5 and 10 mg/L)was 0.1 mg/L.The combined stress of Cu ^(2+)+Pb ^(2+)has an antagonistic effect on G.przewalskii.The expression of Nrf2 was up-regulated under Cu ^(2+),Pb ^(2+)and Cu ^(2+)+Pb ^(2+)combined stress for 12 h.Keap1 was down-regulated in kidneys.The results indicated the negative regulation between Nrf2-Keap1 and Cu ^(2+)+Pb ^(2+)combined stress.The expression of MT was down-regulated in the gill and liver,and the expression of TNF-αwas up-regulated in the gill.Under Cu ^(2+)+Pb ^(2+)combined stress,the expression of TNF-αwas up-regulated.Under Cu ^(2+)and Pb ^(2+)stress,the expressions of Hsp90,Hsp70,iNOS and COX-2 were down-regulated.The results showed that all concentrations of Cu ^(2+),Pb ^(2+)and Cu ^(2+)+Pb ^(2+)had toxic effects on G.przewalskii,and all genes responded to the impact of low concentrations of heavy metal ions.

关 键 词：青海湖裸鲤(Gymnocypris przewalskii) 重金属胁迫 毒性效应 抗氧化应激 基因表达 

分 类 号：S949[农业科学—水产养殖]