镉对单环刺螠非特异性免疫及组织蓄积的影响
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摘要
为探究镉(Cd)对海洋动物的毒性效应,考察了不同质量浓度(0.005、0.05和0.5 mg·L~(-1)) Cd处理对单环刺螠相关免疫生理指标及主要组织中Cd蓄积量的影响。结果表明:Cd胁迫下,血细胞密度(density of hemocyte, DHC)先升后降再升,酚氧化酶(phenoloxidase, PO)活力先升后降,溶菌酶(lysozyme, LSZ)活力先降后升再降,至处理96 h,处理组DHC均显著高于对照组(P<0.05),总蛋白含量变化不显著;除低浓度胁迫组(0.005 mg·L~(-1))PO活力与对照组相近外,其他处理组的PO和LSZ活力均显著低于对照组(P<0.05)。超氧化物歧化酶(superoxide dismutase, SOD)活力和丙二醛(malondialdehyde, MDA)含量总体呈上升趋势,过氧化氢(H_2O_2)含量先升后降再升,至胁迫96 h,各处理组SOD活力、MDA和H_2O_2含量均显著高于对照组(P<0.05)。总之,Cd胁迫对单环刺螠非特异性免疫影响显著,Cd的短期胁迫可刺激DHC、PO和LSZ升高,免疫力增强,而高浓度Cd的胁迫时间较长会造成机体解毒能力下降,持续刺激机体产生过氧化反应,MDA含量升高,可作为单环刺螠在Cd长期胁迫下的免疫检测指标。不同胁迫浓度组单环刺螠肌肉和消化道中的Cd含量随胁迫时间均呈上升趋势,且存在显著的时间-剂量-效应关系(P<0.05),其蓄积量水平为:消化道>肌肉,具有组织特异性。
In this study, the effects of various concentrations of cadmium(Cd) on non-specific immunity indexes,its bioaccumulation and toxicity in the tissues of Urechis unicinctus were investigated. Results show that the density of hemocyte(DHC) initially increased, then decreased before increasing again under Cd stress. In addition, the activity of phenoloxidase(PO) was found to be increased initially and then decreased, while lysozyme(LSZ) activity increased after initial decrement and then decreased. After treatment of 96 h, DHC of the treatments was found to be significantly higher than control group(P<0.05), with no significant change in total protein content. PO activity under low Cd concentration(0.005 mg·L~(-1)) was similar to the control group. In contrast, PO and LSZ activities of other treatments were significantly lower than control. While, superoxide dismutase(SOD) activity and malondialdehyde(MDA) content increased continually, H_2 O_2 content was found to be decreased after initial increment before increasing again. SOD activity, MDA and H_2 O_2 content in treatments were significantly higher than control group(P<0.05) after 96 h of Cd stress. In conclusion, Cd stress exhibited significant effect on non-specific immunity of U. unicinctus. After being subjected to short-term of Cd stress, some immunity indexes, including DHC, and the activities of PO and LSZ were found to be increased, indicating improved immunity. However, long-term exposure of higher Cd concentration led to the decrease of detoxification ability and thus stimulated the body to undergo peroxidation. This led to the increase in MDA content that could be used as an immunoassay indicator of U. unicinctus under long-term Cd stress. Cd content in the muscle and digestive tract of U. unicinctus were found to be increasing with the prolongation of stress time, exhibiting significant time-dose-effect relationship( P <0.05). Moreover, the accumulation of Cd in U. unicinctus showed tissue specificity, with significantly higher Cd content in digestive tract when compared to that of muscle.
In this study, the effects of various concentrations of cadmium(Cd) on non-specific immunity indexes,its bioaccumulation and toxicity in the tissues of Urechis unicinctus were investigated. Results show that the density of hemocyte(DHC) initially increased, then decreased before increasing again under Cd stress. In addition, the activity of phenoloxidase(PO) was found to be increased initially and then decreased, while lysozyme(LSZ) activity increased after initial decrement and then decreased. After treatment of 96 h, DHC of the treatments was found to be significantly higher than control group(P<0.05), with no significant change in total protein content. PO activity under low Cd concentration(0.005 mg·L~(-1)) was similar to the control group. In contrast, PO and LSZ activities of other treatments were significantly lower than control. While, superoxide dismutase(SOD) activity and malondialdehyde(MDA) content increased continually, H_2 O_2 content was found to be decreased after initial increment before increasing again. SOD activity, MDA and H_2 O_2 content in treatments were significantly higher than control group(P<0.05) after 96 h of Cd stress. In conclusion, Cd stress exhibited significant effect on non-specific immunity of U. unicinctus. After being subjected to short-term of Cd stress, some immunity indexes, including DHC, and the activities of PO and LSZ were found to be increased, indicating improved immunity. However, long-term exposure of higher Cd concentration led to the decrease of detoxification ability and thus stimulated the body to undergo peroxidation. This led to the increase in MDA content that could be used as an immunoassay indicator of U. unicinctus under long-term Cd stress. Cd content in the muscle and digestive tract of U. unicinctus were found to be increasing with the prolongation of stress time, exhibiting significant time-dose-effect relationship( P <0.05). Moreover, the accumulation of Cd in U. unicinctus showed tissue specificity, with significantly higher Cd content in digestive tract when compared to that of muscle.
引文
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[25]张林宝,吴惠丰,孙伟,等.菲律宾蛤仔对镉、铜暴露的蓄积作用及其抗氧化酶系统的响应研究[J].南方水产科学,2013,9(5):64-70Zhang B L,Wu H F,Sun W,et al.Bioaccumulation and antioxidant responses in Manila clam(Ruditapes philippinarum)under cadmium and copper exposure[J].South China Fisheries Science,2013,9(5):64-70(in Chinese)
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[2]刘海梅,王苗苗,赵芹.单环刺螠体壁肌酶解产物的营养评价及其溶解性分析[J].食品与机械,2016(5):20-23Liu H M,Wang M M,Zhao Q.Nutritional evaluation and solubility analysis of hydrolysate from Urechis unicinctus body wall muscle[J].Food&Machinery,2016(5):20-23(in Chinese)
[3]Bi Q Q,Han B Q,Feng Y L,et al.Antithrombotic effects of a newly purified fibrinolytic protease from Urechis unicinctus[J].Thrombosis Research,2013,132(2):135-144
[4]Ma X Y,Liu X L,Zhou D,et al.The NF-κB pathway participates in the response to sulfide stress in Urechis unicinctus[J].Fish&Shellfish Immunology,2016,58:229-238
[5]唐永政,宋祥利,翟传阳,等.3种重金属离子对单环刺螠幼螠的急性毒性研究[J].烟台大学学报:自然科学与工程版,2017,30(1):31-35Tang Y Z,Song X L,Zhai C Y,et al.Acute toxicity of three heavy metal ions to Urechis unicinctus juveniles[J].Journal of Yantai University:Natural Science and Engineering Edition,2017,30(1):31-35(in Chinese)
[6]国家环境保护局.GB11607—1989渔业水质标准[S].北京:中国标准出版社,1990National Environmental Protection Agency.GB11607—1989 Water Quality Standard for Fisheries[S].Beijing:China Standard Press,1990(in Chinese)
[7]陈正新,李绍全,王保军,等.青岛地区镉的分布特征及环境效应[A]//中国地质调查局.海岸带地质环境与城市发展研讨会论文集[C].北京:中国大地出版社,2005:271-276Chen Z X,Li S Q,Wang B J,et al.Distribution characteristics and environmental effects of cadmium in Qingdao area[A]//China Geological Survey.Proceedings of the Symposium on Coastal Geological Environment and Urban Development[C].Beijing:China Land Press,2005:271-276(in Chinese)
[8]沈平,陈向东.微生物学实验[M].北京:高等教育出版社,2007:51-53Shen P,Chen X D.Microbiology Experiment[M].Beijing:Higher Education Press,2007:51-53(in Chinese)
[9]王顺昌,许立.不同盐度下中华绒螯蟹血清总蛋白含量和血蓝蛋白含量的变化[J].淮南师范学院学报,2003(3):24-26Wang S C,Xu L.Changes of serum total protein content and hemocyanin content in Eriocheir sinensis under different salinities[J].Journal of Huainan Normal University,2003(3):24-26(in Chinese)
[10]Ashida M.Purification and characterization of pre-phenoloxidase from hemolymph of the silkworm Bombyx mori[J].Archives of Biochemistry&Biophysics,1971,144(2):749-762
[11]Hultmark D,Steiner H,Rasmuson T,et al.Insect immunity.Purification and properties of three inducible bactericidal proteins from hemolymph of immunized pupae of Hyalophora cecropia[J].European Journal of Biochemistry,2010,106(1):7-16
[12]Yamuna A,Saravana B P,Geraldine P.Effects of Hg and Cu on hemocytes-mediated functions in the prawn,Macrobrachium malcolmsonii[J].Journal of Environmental Biology,2002,23(1):7-13
[13]蒋云霞,徐华,艾春香.镉或铬胁迫对拟穴青蟹血细胞总数和酚氧化酶活性的影响[J].海洋通报,2010,29(6):649-653Jiang Y X,Xu H,Ai C X.Effects of Cd2+or Cr6+stress on the THC and PO activity of mud crab Scylla paramamosain[J].Ocean Report,2010,29(6):649-653(in Chinese)
[14]Lorenz S,Francese M,Smith V J,et al.Heavy metals affect the circulating haemocyte number in the shrimp Palaemon elegans[J].Fish&Shellfish Immunology,2001,11(6):459-472
[15]Zahedi S,Mirvaghefi A,Rafati M,et al.Cadmium accumulation and biochemical parameters in juvenile Persian sturgeon,Acipenser persicus,upon sublethal cadmium exposure[J].Comparative Clinical Pathology,2013,22(5):805-813
[16]张婷,张玉,李德亮,等.鲢鱼对环境镉胁迫的生理响应及其对湘江鱼类增殖放流的启示[J].中国科学:生命科学,2013,43(5):432-439Zhang T,Zhang Y,Li D L,et al.Exposure of silver carp(Hypophthalmichthys molitrix)to environmentally relevant levels of cadmium:Hematology,muscle physiology,and implications for stock enhancement in the Xiangjiang River(Hunan,China)[J].Science China:Life Science,2013,43(5):432-439(in Chinese)
[17]王轶南,穆晓虎,封妮莎,等.仿刺参体腔液中酚氧化酶活性的分析[J].大连水产学院学报,2013,28(4):319-322Wang Y N,Mu X H,Feng N S,et al.Analysis of phenoloxidase activity in the coelomic fluid of sea cucumber Apostichopus japonicus[J].Journal of Dalian Ocean University,2013,28(4):319-322(in Chinese)
[18]秦圣娟,秦琴,金芬芬,等.镉对长江华溪蟹免疫相关酶活性的影响[J].四川动物,2011,30(5):765-767Qin S J,Qin Q,Jin F F,et al.Effects of cadmium on enzymes related to immune in freshwater crab Sinopotamon yangtsekiense[J].Sichuan Journal of Zoology,2011,30(5):765-767(in Chinese)
[19]Wang J,Yang X P,Wang J H,et al.Probing the binding interaction between cadmium(II)chloride and lysozyme[J].New Journal of Chemistry,2016,40(4):3738-3746
[20]李慧,狄桂兰,裴超,等.镉胁迫下鲤血液免疫指标的变化[J].水生态学杂志,2017,38(4):90-95Li H,Di G L,Pei C,et al.Immune response of common carp(Cyprinus carpio)exposed to cadmium[J].Journal of Hydroecology,2017,38(4):90-95(in Chinese)
[21]王志,武二栓.水环境镉对鲤鱼免疫毒性状况的分析[J].现代农业,2005(1):4-5Wang Z,Wu E S.Analysis on the immunotoxicity of cadmium in water environment to Cyprinus carpio[J].Modern Agricuture,2005(1):4-5(in Chinese)
[22]朱秀敏.超氧化物歧化酶的生理活性[J].当代医学,2011,17(15):26-27Zhu X M.Physiological activity of superoxide dismutase[J].Contemporary Medicine,2011,17(15):26-27(in Chinese)
[23]Li Y Q,Wang L,Liu N,et al.Effects of cadmium on enzyme activity and lipid peroxidation in freshwater crab Sinopotamon yangtsekiense[J].Acta Hydrobiologica Sinica,2008,32(3):373-379
[24]吴天利,李广丽,师尚丽,等.镉胁迫对凡纳滨对虾血清中一氧化氮合成酶和超氧化物歧化酶活性的影响[J].热带海洋学报,2008,27(6):62-65Wu T L,Li G L,Shi S L,et al.Effects of cadmium stress on NOS and SOD activities in Litopeneaus vannamei[J].Journal of Tropical Oceanography,2008,27(6):62-65(in Chinese)
[25]张林宝,吴惠丰,孙伟,等.菲律宾蛤仔对镉、铜暴露的蓄积作用及其抗氧化酶系统的响应研究[J].南方水产科学,2013,9(5):64-70Zhang B L,Wu H F,Sun W,et al.Bioaccumulation and antioxidant responses in Manila clam(Ruditapes philippinarum)under cadmium and copper exposure[J].South China Fisheries Science,2013,9(5):64-70(in Chinese)
[26]Tamagno E,Robino G,Obbili A,et al.H2O2and 4-hydroxynonenal mediate amyloid beta-induced neuronal apoptosis by activating JNKs and p38MAPK[J].Experimental Neurology,2003,180(2):144-155
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