Cd^(2+)和Zn^(2+)胁迫对青海湖裸鲤抗氧化系统和非特异性免疫机能的影响  

Effects of Cd^(2+) and Zn^(2+) Stress on Antioxidant System and Non-specific Immune Function in Gymnocypris przewalskii

作　　者：王蓉 李长忠 赵静 马淑兄 贾春艳 勾化宇 李兰英 陈艳霞 祁洪芳[3] 金文杰 WANG Rong;LI Chang-Zhong;ZHAO Jin;MA Shu-Xiong;JIA Chun-Yan;GOU Hua-Yu;LI Lan-Ying;CHEN Yan-Xia;QI Hong-Fang;JINWen-Jie(College of Eco-Environmental Engineering,Qinghai University,Xining 810016,China;Key Laboratory of Plateau Cold-water Fish Culture and Eco-environmental Conservation(Co-construction by Ministry and Province),Ministry of Agriculture and Rural Affairs,Xining 810016,China;Key Laboratory of Breeding and Protection of Gymnocypris przewalskii,Rescue Center of Gymnocypris przewalskii,Xining 810016,China)

机构地区：[1]青海大学生态环境工程学院,西宁810016 [2]农业农村部高原冷水鱼养殖与生态环境保护重点实验室(部省共建),西宁810016 [3]青海湖裸鲤救护中心,青海省青海湖裸鲤繁育与保护重点实验室,西宁810016

出　　处：《农业生物技术学报》2024年第12期2808-2821,共14页Journal of Agricultural Biotechnology

基　　金：青海省科协中青年科技人才托举工程(2022QHSKXRCTJ34);国家自然科学基金项目(31460700)。

摘　　要：青海湖裸鲤(Gymnocypris przewalskii)是一类特有的耐低温耐盐碱性高原裂腹鱼类,重金属胁迫是限制其溯河洄游产卵繁育的主要非生物胁迫之一。为探讨Cd^(2+)和Zn^(2+)胁迫对青海湖裸鲤的急性毒性作用、抗氧化能力及非特异性免疫机能的影响,本研究采用静水生物测定法确定不同浓度Cd^(2+)和Zn^(2+)胁迫青海湖裸鲤24、48、72和96 h的半致死浓度(lethal concentration for 50%,LC_(50))及安全浓度(safe concentration,SC);通过酶标仪测定Cd^(2+)和Zn^(2+)胁迫后其鳃、肾脏和肝脏组织中超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)、酸性磷酸酶(acid phosphatase,ACP)和碱性磷酸酶(alkaline phosphatase,AKP)的活性,并用qPCR检测NF-κB相关免疫基因转化生长因子β激活激酶1(transforming growth factorβ-activatrd kinase 1,TAK1)、IκB抑制因子激酶α(inhibitory nuclear factor kinase-κBα,IKKα)、核因子κB抑制因子α(inhibitor nuclear factor-κBα,IκBα)、核转录因子κB(nuclear transcription factor-κB,NF-κB)、白细胞介素8(interleukin 8,IL-8)、锌指蛋白A20(A20)和肿瘤坏死因子-α(tumor necrosis factorα,TNF-α)的相对表达量。结果显示,Cd^(2+)胁迫青海湖裸鲤幼鱼24、48、72和96 h的LC_(50)分别为1.455、1.022、0.579和0.428 mg/L;Zn^(2+)胁迫青海湖裸鲤幼鱼24、48、72和96 h的LC_(50)分别为4.634、2.797、1.843和1.157 mg/L;Cd^(2+)和Zn^(2+)的SC分别为0.0043和0.1157 mg/L,毒性大小为Cd^(2+)>Zn^(2+)。青海湖裸鲤的肾脏和肝脏中SOD和CAT活性对Cd^(2+)和Zn^(2+)胁迫有不同程度的响应,但较高浓度Cd^(2+)胁迫24 h时鳃组织中SOD和CAT的活性较高;胁迫后的鳃和肾脏中ACP活性显著降低(P<0.05),鳃和肝脏中AKP活性显著降低(P<0.05);在鳃、肾脏和肝脏中TAK1、IκBα、NF-κB和IL-8的表达量显著上调(P<0.05),而IKKα、A20和TNF-α表达量相对较低,尤其在肝脏中表现更为明显。由此可知,Cd^(2+)和Zn^(2+)胁迫均可诱导青Gymnocypris przewalskii is a kind of Schizothoracinae with low temperature and salt and alkali resistance.Heavy metal stress is one of the main abiotic stresses that limit the anadromous migration and breeding of G.przewalskii.To investigate the effects of Cd^(2+)and Zn^(2+)stress on the acute toxicity,antioxidant ability and non-specific immune function of G.przewalskii.G.przewalskii was exposed to different concentrations of Cd^(2+)and Zn^(2+)at 24,48,72,and 96 h.The semi-lethal concentration for 50%(LC_(50))and the safe concentration(SC)were determined by static bioassay.The activities of superoxide dismutase(SOD),catalase(CAT),acid phosphatase(ACP),and alkaline phosphatase(AKP)in gills,kidney and liver tissues of G.przewalskii were detected by the automatic microplate reader,and the relative expression of NF-κB related immune genes,such as transforming growth factorβ-activatrd kinase 1(TAK1),inhibitory nuclear factor kinase-κBα(IKKα),inhibitor nuclear factor-κBα(IκBα),nuclear transcription factor-κB(NF-κB),interleukin 8(IL-8),zinc finger protein A20(A20),and tumor necrosis factorα(TNF-α)were detected by qPCR.The results showed that the LC_(50)of G.przewalskii under Cd^(2+)stress for 24,48,72,and 96 h were 1.455,1.022,0.579,and 0.428 mg/L,respectively.The LC_(50)of G.przewalskii under Zn^(2+)stress for 24,48,72,and 96 h were 4.634,2.797,1.843,and 1.157 mg/L,respectively.The SC of Cd^(2+)and Zn^(2+)were 0.0043 and 0.1157 mg/L,respectively,and the toxicity magnitude was Cd^(2+)>Zn^(2+).After Cd^(2+)and Zn^(2+)stress,the activities of SOD and CAT in the kidney and liver of G.przewalskii shown varying degrees of response to Cd^(2+)and Zn^(2+)stress,but in high concentration,the activities were higher at 24 h in the gills.The activities of ACP were significantly reduced(P<0.05)in the gills and kidney,and the activities of AKP were significantly decreased(P<0.05)in the gills and liver.The expression of TAK1,IκBα,NF-κB,and IL-8 were highly up regulated(P<0.05)in gills,kidney,and liver,while the expres

关 键 词：青海湖裸鲤 重金属胁迫 急性毒性 抗氧化能力 免疫机能 

分 类 号：S917.4[农业科学—水产科学]