铜对斑马鱼鳃的损伤及其作用机制
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摘要
目的探讨铜对斑马鱼鳃的生物毒性作用及其机制。方法设置0.05 mg/L、0.1 mg/L、0.2 mg/L 3个铜离子暴露浓度和1个对照组,研究铜离子对斑马鱼鳃组织显微和超微结构,抗氧化防御系统超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量及免疫相关因子肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平的影响。结果铜离子暴露组斑马鱼鳃均出现不同程度的损伤,中低浓度组以防御损伤为主,高浓度组以直接损伤为主。光学显微镜下可看到上皮细胞肿胀、脱落,表面大量黏液,鳃小片基部细胞增生甚至融合;透射电子显微镜下可观察到暴露组线粒体、粗面内质网等细胞器肿胀,线粒体嵴崩落或消失等细胞胀亡形态特征;SOD活性和MDA含量随铜离子浓度增加和暴露时间的延长出现不同程度的上升趋势,铜离子暴露对斑马鱼SOD、MDA为诱导效应;与对照组相比,不同时间、不同浓度重金属铜处理后TNF-α、IL-6、IL-1β水平出现不同程度的上升趋势。结论重金属铜对斑马鱼鳃组织结构和抗氧化系统有明显的损伤,能够诱导免疫炎症反应。
Objective To study the effect of copper on microstructures and ultrastructures of gill of zebrafish,antioxidant defense system of superoxide dismutase( SOD) activity,malondialdehyde( MDA) content and related immune factors [tumor necrosis factor alpha( TNF-alpha),interleukin( IL)-6,IL-1 beta) ]in the acute toxicity test. Methods Three copper exposures( 0. 05 mg/L,0. 1 mg/L,0. 2 mg/L) and a control group were set in this study. Results The copper exposure groups showed varying degrees of damage,such as the epithelial cell swelling,shedding,surface mucus,basal cell hyperplasia and gill lamellae fusion under the light microscope. The mitochondria and rough endoplasmic reticulum cell were swelled,and mitochondrial cristae collapse or the disappearance of oncosis morphology were observed under transmission electron microscope( TEM). SOD activity and MDA content extended upward as increase of the copper ion concentration and exposure time. In addition,copper ion exposure on zebrafish SOD,MDA as compared with the control,showed an induction effect. Compared to the control group,TNF-alpha,IL-6,and IL-1 beta levels increased by different extents after treatment with different concentrations of copper and time. Conclusion The heavy metal copper has obvious damage and immune toxicity to the gill structure and antioxidant system of zebrafish.
Objective To study the effect of copper on microstructures and ultrastructures of gill of zebrafish,antioxidant defense system of superoxide dismutase( SOD) activity,malondialdehyde( MDA) content and related immune factors [tumor necrosis factor alpha( TNF-alpha),interleukin( IL)-6,IL-1 beta) ]in the acute toxicity test. Methods Three copper exposures( 0. 05 mg/L,0. 1 mg/L,0. 2 mg/L) and a control group were set in this study. Results The copper exposure groups showed varying degrees of damage,such as the epithelial cell swelling,shedding,surface mucus,basal cell hyperplasia and gill lamellae fusion under the light microscope. The mitochondria and rough endoplasmic reticulum cell were swelled,and mitochondrial cristae collapse or the disappearance of oncosis morphology were observed under transmission electron microscope( TEM). SOD activity and MDA content extended upward as increase of the copper ion concentration and exposure time. In addition,copper ion exposure on zebrafish SOD,MDA as compared with the control,showed an induction effect. Compared to the control group,TNF-alpha,IL-6,and IL-1 beta levels increased by different extents after treatment with different concentrations of copper and time. Conclusion The heavy metal copper has obvious damage and immune toxicity to the gill structure and antioxidant system of zebrafish.
引文
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[12]Ioannidis O,Papaziogas B,Tsiaousis P,et al.Effect of moxifloxacin on survival,lipid peroxidation and inflammation in immunosuppressed rats with soft tissue infection caused by Stenotrophomonas maltophilia[J].Microbiol Immunol,2014,58(2):96-102.
[13]Wang Ch X.Study on the effects of copper to the toxicity of the Yellow River Carp[D].Henan Agricultural University,2010.(in Chinese)王春秀.水体铜对黄河鲤毒性作用的研究[D].河南农业大学,2010.
[14]Roh YS,Cho A,Islam MR,et al.3,3’-Diindolylmethane induces immunotoxicity via splenocyte apoptosis in neonatal mice[J].Toxicol Lett,2011,206(2):218-228.
[15]Prieto-Alamo MJ,Abril N,Osuna-Jiménez I,et al.Solea senegalensis genes responding to lipopolysaccharide and copper sulphate challenges:large-scale identification by suppression subtractive hybridization and absolute quantification of transcriptional profiles by real-time RT-PCR[J].Aquat Toxicol,2009,91(4):312-319.
[16]Geist J,Werner I,Eder KCGJ,et al.Comparisons of tissuespecific transcription of stress response genes with whole animal endpoints of adverse effect in striped bass(Morone saxatilis)following treatment with copper and esfenvalerate[J].Aquat Toxicol,2007,85(1):28-39.
[2]Chen Zh L.Application of zebrafish in drug screens for active ingredients[J].China Journal of Chinese Materia Medica,2015,40(7):1235-1239.(in Chinese)陈志亮.斑马鱼在药物筛选中的应用[J].中国中药杂志,2015,40(7):1235-1239.
[3]Boyle D,Al-Bairuty GA,Ramsden CS,et al.Subtle alterations in swimming speed distributions of rainbow trout exposed to titanium dioxide nanoparticles are associated with gill rather than brain injury[J].Aquat Toxicol,2013,126(2):116-127.
[4]Monteiro SM,Rocha E,Mancera JM,et al.A stereological study of copper toxicity in gills of Oreochromis niloticus[J].Ecotoxicol Environ Saf,2009,72(1):213-223.
[5]Cengiz EI,Unlu E.Sublethal effects of commercial deltamethrin on the structure of the gill,liver and gut tissues of mosquitofish,Gambusia affinis:A microscopic study[J].Environ Toxicol Pharmacol,2006,21(3):246-253.
[6]Mitra S,Keswani T,Ghosh N,et al.Copper induced immunotoxicity promote differential apoptotic pathways in spleen and thymus[J].Toxicology,2013,306(12):74-84.
[7]Weerasinghe P,Buja LM.Oncosis:an important non-apoptotic mode of cell death[J].Exp Mol Pathol,2012,93(3):302-308.
[8]Xu YJ,Liu XZh,Ma AJ.Current research on toxicity effect and molecular mechanism of heavy metals on fish[J].Marine Sciences,2004,28(10):67-70.(in Chinese)徐永江,柳学周,马爱军.重金属对鱼类毒性效应及其分子机理的研究概况[J].海洋科学,2004,28(10):67-70.
[9]Huang W,Cao LA,Ye ZJ,et al.Antioxidative responses and bioaccumulation in Japanese flounder larvae and juveniles under chronic mercury exposure[J].Comp Biochem Physiol C Toxicol Pharmacol,2010,152(1):99-106.
[10]Lin CT,Tseng WC,Hsiao NW,et al.Characterization,molecular modelling and developmental expression of zebrafish manganese superoxide dismutase[J].Fish Shellfish Immunol,2009,27(2):318-324.
[11]Ma TW,Zhu Ch,Zhou K,et al.Effects of Cd,Pb and combined contaminated sediments on hepatopancreatic SOD and MT in Bellamya aeruginosa.[J].Journal of Agro-Environment Science,2010,29(1):30-37.(in Chinese)马陶武,朱程,周科,等.Cd Pb单一及复合污染沉积物对铜锈环棱螺肝胰脏SOD和MT的影响[J].农业环境科学学报,2010,29(1):30-37.
[12]Ioannidis O,Papaziogas B,Tsiaousis P,et al.Effect of moxifloxacin on survival,lipid peroxidation and inflammation in immunosuppressed rats with soft tissue infection caused by Stenotrophomonas maltophilia[J].Microbiol Immunol,2014,58(2):96-102.
[13]Wang Ch X.Study on the effects of copper to the toxicity of the Yellow River Carp[D].Henan Agricultural University,2010.(in Chinese)王春秀.水体铜对黄河鲤毒性作用的研究[D].河南农业大学,2010.
[14]Roh YS,Cho A,Islam MR,et al.3,3’-Diindolylmethane induces immunotoxicity via splenocyte apoptosis in neonatal mice[J].Toxicol Lett,2011,206(2):218-228.
[15]Prieto-Alamo MJ,Abril N,Osuna-Jiménez I,et al.Solea senegalensis genes responding to lipopolysaccharide and copper sulphate challenges:large-scale identification by suppression subtractive hybridization and absolute quantification of transcriptional profiles by real-time RT-PCR[J].Aquat Toxicol,2009,91(4):312-319.
[16]Geist J,Werner I,Eder KCGJ,et al.Comparisons of tissuespecific transcription of stress response genes with whole animal endpoints of adverse effect in striped bass(Morone saxatilis)following treatment with copper and esfenvalerate[J].Aquat Toxicol,2007,85(1):28-39.