铜对草鱼生长及肾脏中免疫相关基因表达的影响
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摘要
为了探讨水体重金属铜对草鱼(Ctenopharyngodon idellus)的生长和免疫的毒性作用,分别将草鱼在不同质量浓度的Cu~(2+)溶液中暴露30、60和90 d,检测了草鱼的均重和增重率的变化,以及草鱼肾脏中6种免疫相关基因表达的变化.结果表明:与对照组相比,各质量浓度Cu~(2+)暴露均使草鱼在30、60和90 d后的均体质和增体质率下降;0.40和0.60 mg·L~(-1)质量浓度Cu~(2+)暴露60和90 d后,IL-1β,CCL4和TNF-α基因在草鱼肾脏中的表达量均极显著升高(P<0.01),而IFN-γ和IgM基因在草鱼肾脏中的表达量均显著或极显著下降(P<0.05或P<0.01);Cu~(2+)暴露30、60和90 d后,草鱼肾脏中MT基因表达量均随着Cu~(2+)质量浓度的增加先升高后下降,在Cu~(2+)质量浓度为0.10 mg/L时达到最大值.综上,高质量浓度和长时间的Cu~(2+)暴露阻碍了草鱼的生长,引起了草鱼肾脏细胞的炎症反应,降低了草鱼的免疫功能.
To investigate the toxic effect of waterborne copper on the growth and immunity of grass carp(Ctenopharyngodon idellus), grass carps were exposed in different concentrations of Cu~(2+)solutions for 30, 60 and 90 days, then the weight and weight gain rate, the expression changes of six immunity-related genes in kidney of grass carps were measured. The results showed that: Compared with the control group, the average weights and weight gain rates all decreased under different concentrations of Cu~(2+)exposures for 30, 60, and 90 days; After exposure to 0.40 and 0.60 mg·L~(-1) Cu~(2+)for 60 and 90 days, the expression levels of IL-1β, CCL4, and TNF-α genes in kidney of grass carp were extremely significantly increased(P<0.01), however, the expression levels of IFN-γ and IgM genes were significantly or extremely significantly decreased(P<0.05 or P<0.01); After 30, 60 and 90 days of Cu~(2+)exposures, the MT gene expression in kidney of grass carp increased at first and then decreased with the increasing of Cu~(2+)concentration, and reached the maximum value at the Cu~(2+)concentration of 0.10 mg·L~(-1). In conclusion,high concentration and long time Cu~(2+)exposure hindered the growth of grass carp, caused inflammatory reaction of kidney cells, and weakened the immune function of grass carp.
To investigate the toxic effect of waterborne copper on the growth and immunity of grass carp(Ctenopharyngodon idellus), grass carps were exposed in different concentrations of Cu~(2+)solutions for 30, 60 and 90 days, then the weight and weight gain rate, the expression changes of six immunity-related genes in kidney of grass carps were measured. The results showed that: Compared with the control group, the average weights and weight gain rates all decreased under different concentrations of Cu~(2+)exposures for 30, 60, and 90 days; After exposure to 0.40 and 0.60 mg·L~(-1) Cu~(2+)for 60 and 90 days, the expression levels of IL-1β, CCL4, and TNF-α genes in kidney of grass carp were extremely significantly increased(P<0.01), however, the expression levels of IFN-γ and IgM genes were significantly or extremely significantly decreased(P<0.05 or P<0.01); After 30, 60 and 90 days of Cu~(2+)exposures, the MT gene expression in kidney of grass carp increased at first and then decreased with the increasing of Cu~(2+)concentration, and reached the maximum value at the Cu~(2+)concentration of 0.10 mg·L~(-1). In conclusion,high concentration and long time Cu~(2+)exposure hindered the growth of grass carp, caused inflammatory reaction of kidney cells, and weakened the immune function of grass carp.
引文
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[2] Eyckmans M,Celis N,Horemans N,et al.Exposure to waterborne copper reveals differences in oxidative stress response in three freshwater fish species[J].Aquatic Toxicology,2011,103:112-120.
[3] 刘福军,张饮江,王明学.铜对鱼类慢性毒性研究进展[J].水生生物学报,2003,27(3):302-307.
[4] 王利,汪开毓.铜在鲤体内的蓄积及毒性的研究[J].淡水渔业,2008,38(4):45-48.
[5] 任洪涛,张春暖,林霖.Mo6+对草鱼组织器官及抗氧化酶活性的影响[J].水产科学,2017,36(3):317-322.
[6] Zhu Qinglin,Luo Zhi,Zhuo Meiqin,et al.In vitro exposure to copper influences lipid metabolism in hepatocytes from grass carp (Ctenopharyngodon idellus)[J].Fish Physiology and Biochemistry,2014,40(2):595-695.
[7] Wang Wanbin,Chen Sha,Wu Min,et al.Predicting copper toxicity to Hypophthalmichthys molitrix and Ctenopharyngodon idellus based on biotic ligand model[J].Environmental Sciences,2014,35(10):3947-3951.
[8] 朱玉娇,刘永,胡成钰.Pb2+胁迫对草鱼过氧化氢酶和髓过氧化物酶的影响[J].南昌大学学报(理科版),2012,36(2):176-179.
[9] 华涛,周启星.Cd-Zn对草鱼(Ctenopharyngodon idellus)的联合毒性及对肝脏超氧化物歧化酶(SOD)活性的影响[J].环境科学学报,2009,29(3):600-606.
[10] Hansen JA,Lipton J,Welsh PG,et al.Relationship between exposure duration,tissue residues,growth,and mortality in rainbow trout (Oncorhynchus mykiss) juveniles sub-chronically exposed to copper[J].Aquatic Toxicology,2002,58:175-188.
[11] James R,Sampath K,Jothilakshmi S,et al.Effects of copper toxicity on growth,reproduction and metal accumulation in chosen ornamental fishes[J].Ecohydrology and Hydrobiology,2008,8(1):89-97.
[12] Liu XJ,Luo Z,Xiong BX,et al.Effect of waterborne copper exposure on growth,hepatic enzymatic activities and histology in Synechogobius hasta[J].Ecotoxicology and Environmental Safety,2010,73:1286-1291.
[13] Jiang Hongxia,Kong Xianghui,Wang Shuping,et al.Effect of copper on growth,digestive and antioxidant enzyme activities of juvenile Qihe crucian carp,Carassius carassius,during exposure and recovery[J].Bulletin of Environmental Contamination and Toxicology,2016,96:333-340.
[14] Bird Steve,Zou Jun,Wang Tiehui,et al.Evolution of interleukin-1β[J].Cytokine Growth Factor Reviews,2002,13:483-502.
[15] Glass W G,Rosenberg H F,Murphy P M.Chemokine regulation of inflammation during acute viral infection[J].Current Opinion in Allergy and Clinical Immunology,2003,3(6):467-473.
[16] Ebert L M,Scharrli P,Moser B.Chemokine-mediated control of T cell traffic in lymphoid and peripheral tissues[J].Molecular Immunology,2005,42(7):799-809.
[17] Horiuchi T,Mitoma H,Harashima S,et al.Transmembrane TNF-α:structure,function and interaction with anti-TNF agents[J].Rheumatology,2010,49(7):1215-1228.
[18] N rregaard R D,Dang M,Bach L,et al.Comparison of heavy metals,parasites and histopathology in sculpins (Myoxocephalus spp.) from two sites at a lead-zinc mine in North East Greenland[J].Environmental Research,2018,165:306-316.
[19] Khan M I,Khisroon M,Khan A,et al.Bioaccumulation of heavy metals in water,sediments,and tissues and their histopathological effects on Anodonta cygnea (Linea,1876) in Kabul River,Khyber Pakhtunkhwa,Pakistan[J].BioMed Research International,2018,6:1-10.
[20] Yin Jian,Wang Aiping,Li Wanfang,et al.Time-response characteristic and potential biomarker identification of heavy metal induced toxicity in zebrafish[J].Fish and Shellfish Immunology,2018,72:309-317.
[21] 毛明光,刘宗柱,张培军.鱼类干扰素功能及信号转导研究[J].海洋科学,2008,32(2):85-90.
[22] Reddy P S,Corley R B.The contribution of ER quality control to the biologic functions of secretory IgM[J].Immunology Today,1999,20:582-588.
[23] Klimovich V B,Samoilovich M P,Klimovich B V.Problem of J-chain of immunoglobulins[J].Journal of Evolutionary Biochemistry and Physiology,2008,44(2):151-166.
[24] Ghazy H A,Abdel-Razek M A S,Nahas A F E,et al.Assessment of complex water pollution with heavy metals and pyrethroid pesticides on transcript levels of metallothionine and immune related genes[J].Fish and Shellfish Immunology,2017,68:318-326.
[25] Brouwer M,Brouwer T H.Biochemical defense mechanisms against copper-induced oxidative damage in the blue crab,Callinectes sapidus[J].Archives of Biochemistry and Biophysics,1998,351(2):257-264.
[26] Hogstrand C,Haux C.Binding and detoxification of heavy metals in lower vertebrates with reference to metallothionein[J].Comparative Biochemistry and Physiology Part C:Comparative Pharmacology,1991,100(1/2):137-141.
[27] Bonham K,Gedamu L.Induction of metallothionein and metallothionein mRNA in rainbow-trout liver following cadmium treatment[J].Bioscience Reports,1984,4(8):633-642.
[28] Olsson P E,Larsson A,Maage A,et al.Induction of metallothionein synthesis in rainbow trout,Salmo gairdneri,during long-term exposure to waterborne cadmium[J].Fish Physiological and Biochemistry,1989,6(4):221-229.
[29] Ren Fei,Jiang Hui,Sun Jiangling,et al.Cloning,characterization,expression,and copper sensitivity of the metallothionein-1 gene in the Chinese mitten crab,Eriocheir sinensis[J].Molecular biology reports,2011,38(4):2383-2393.
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