纳米氧化镍颗粒对长牡蛎(Crassostrea gigas)抗氧化防御体系的影响
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摘要
纳米氧化镍(nNiO)作为一种广泛使用的纳米颗粒,其水生毒理效应研究还很有限。为探索n Ni O对海洋贝类的毒性机制,本研究将长牡蛎(Crassostrea gigas)置于不同浓度(0、1、10、100 mg·L~(-1))的n Ni O中暴露96 h,分别测定鳃和消化腺组织的丙二醛(MDA)含量和超氧化物歧化酶(SOD)、过氧化物酶(POD)以及过氧化氢酶(CAT)活性,并通过实时荧光定量PCR技术测定了鳃和消化腺中应激蛋白HSP70和AOX基因的表达变化。结果显示:在100 mg·L~(-1)n Ni O处理下,2种组织中MDA含量均显著性升高(P<0.01),显示纳米颗粒造成了长牡蛎的脂质过氧化,并可能引起相应的氧化损伤。同时,n Ni O暴露也诱导了长牡蛎抗氧化酶(SOD、CAT和POD)活性的改变。其中,SOD和CAT活性在10 mg·L~(-1)浓度处理组达到最高,而POD活性在1 mg·L~(-1)浓度组即达最高值。在高浓度n Ni O(100 mg·L~(-1))胁迫下,3种抗氧化酶的活性均比低浓度(1和10 mg·L~(-1))处理组降低,表明抗氧化酶的保护作用在较低浓度暴露下更有效;而热激蛋白(hsp70)和交替氧化酶(aox)基因却分别在长牡蛎消化腺和鳃组织中上调表达(P<0.01),并表现出一定的组织差异。说明高浓度纳米颗粒暴露中主要是应激蛋白发挥了作用。本文结果为纳米氧化镍对海洋双壳贝类的毒性机制研究及生态风险评估提供了基础数据。
Nano-nickel oxide(nNiO) is a widely used nanoparticle, and the research on aquatic toxicological effects of n Ni O was limited, especially for marine organisms. To explore the mechanism of toxicity of n Ni O to marine bivalve molluscs, the Pacific oysters(Crassostrea gigas) were exposed to nNiO at different concentrations(0, 1,10, 100 mg·L~(-1)) for 96 h in the present study. The activities of superoxide dismutase(SOD), peroxidase(POD),catalase(CAT) and malondialdehyde(MDA) content in gill and digestive glands were measured. The expression changes of the shock protein HSP70 and AOX gene in gill and digestive gland were also determined by real-time fluorescence quantitative PCR. The results showed that the content of the MDA in both tissues increased significantly(P<0.01) in the 100 mg·L~(-1) treatment group, suggesting that the nanoparticles caused lipid peroxidation of oysters and may cause oxidative damage. Meanwhile, nNiO exposure also induced changes in the antioxidant enzymes(SOD, CAT, and POD) activities of oysters. The SOD and CAT activities reached the maximum in the 10 mg·L~(-1) treatment group, while the POD activity reached the highest in the 1 mg·L~(-1) concentration group. At high concentrations of nNiO(100 mg·L~(-1)) group, the activities of the three antioxidant enzymes were lower than those in lower concentrations(1 and 10 mg·L~(-1)), indicating that the activities of antioxidant enzymes were repressed by higher concentrations of nNiO exposure. However, heat shock protein(hsp70) and alternative oxidase(aox) genes were up-regulated in digestive glands and gills, respectively(P<0.01). It is indicated that the stress protein(HSP70 and AOX) played a key role with higher nNiO exposure concentration. The results provided basic data for the study on the toxicity mechanism of nano-nickel oxides to marine bivalve molluscs.
Nano-nickel oxide(nNiO) is a widely used nanoparticle, and the research on aquatic toxicological effects of n Ni O was limited, especially for marine organisms. To explore the mechanism of toxicity of n Ni O to marine bivalve molluscs, the Pacific oysters(Crassostrea gigas) were exposed to nNiO at different concentrations(0, 1,10, 100 mg·L~(-1)) for 96 h in the present study. The activities of superoxide dismutase(SOD), peroxidase(POD),catalase(CAT) and malondialdehyde(MDA) content in gill and digestive glands were measured. The expression changes of the shock protein HSP70 and AOX gene in gill and digestive gland were also determined by real-time fluorescence quantitative PCR. The results showed that the content of the MDA in both tissues increased significantly(P<0.01) in the 100 mg·L~(-1) treatment group, suggesting that the nanoparticles caused lipid peroxidation of oysters and may cause oxidative damage. Meanwhile, nNiO exposure also induced changes in the antioxidant enzymes(SOD, CAT, and POD) activities of oysters. The SOD and CAT activities reached the maximum in the 10 mg·L~(-1) treatment group, while the POD activity reached the highest in the 1 mg·L~(-1) concentration group. At high concentrations of nNiO(100 mg·L~(-1)) group, the activities of the three antioxidant enzymes were lower than those in lower concentrations(1 and 10 mg·L~(-1)), indicating that the activities of antioxidant enzymes were repressed by higher concentrations of nNiO exposure. However, heat shock protein(hsp70) and alternative oxidase(aox) genes were up-regulated in digestive glands and gills, respectively(P<0.01). It is indicated that the stress protein(HSP70 and AOX) played a key role with higher nNiO exposure concentration. The results provided basic data for the study on the toxicity mechanism of nano-nickel oxides to marine bivalve molluscs.
引文
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[2]石清清,邓代莉,颜椿,等.纳米金属氧化物对土壤酶活性的影响研究进展[J].生态毒理学报, 2018, 13(2):47-56Shi Q Q, Deng D L, Yan C, et al. Review on effects of engineered nano-metal oxide particles to soil enzyme activities[J]. Asian Journal of Ecotoxicology, 2018, 13(2):47-56(in Chinese)
[3] Wang Y L, Ding L, Yao C J, et al. Toxic effects of metal oxide nanoparticles and their underlying mechanisms[J].Science China Materials, 2017, 60(2):93-108
[4] Colvin V L. The potential environmental impact of engineered nanomaterials[J]. Nature Biotechnology, 2003, 21(10):1166-1170
[5] Hoet P H, Nemmar A, Nemery B. Health impact of nanomaterials?[J]. Nature Biotechnology, 2004, 22(1):19
[6] Ahamed M, Ali D, Alhadlaq H A, et al. Nickel oxide nanoparticles exert cytotoxicity via oxidative stress and induce apoptotic response in human liver cells(HepG2)[J].Chemosphere, 2013, 93(10):2514-2522
[7] Salvadori M R, Ando R A, Nascimento C A, et al. Extra and intracellular synthesis of nickel oxide nanoparticles mediated by dead fungal biomass[J]. Plos One, 2015, 10(6):1-15
[8] Bi H, Li S, Zhang Y, et al. Ferromagnetic-like behavior of ultrafine Ni O nanocrystallites[J]. Journal of Magnetism&Magnetic Materials, 2004, 277(3):363-367
[9] Nogueira V, Lopes I, Rochasantos T A, et al. Assessing the ecotoxicity of metal nano-oxides with potential for wastewater treatment[J]. Environmental Science&Pollution Research, 2015, 22(17):13212-13224
[10] Oukarroum A, Barhoumi L, Samadani M, et al. Toxic effects of nickel oxide bulk and nanoparticles on the aquatic plant Lemna gibba L.[J]. Biomed Research International, 2015, 2015:501326
[11] Capasso L, Camatini M, Gualtieri M. Nickel oxide nanoparticles induce inflammation and genotoxic effect in lung epithelial cells[J]. Toxicology Letters, 2014, 226(1):28-34
[12] Siddiqui M A, Ahamed M, Ahmad J, et al. Nickel oxide nanoparticles induce cytotoxicity, oxidative stress and apoptosis in cultured human cells that is abrogated by the dietary antioxidant curcumin[J]. Food&Chemical Toxicology, 2012, 50(3-4):641-647
[13] Horie M, Fukui H, Endoh S, et al. Comparison of acute oxidative stress on rat lung induced by nano and finescale, soluble and insoluble metal oxide particles:Ni O and Ti O2[J]. Inhalation Toxicology, 2012, 24(7):391-400
[14]谢嘉.典型重金属(Cd2+、Pb2+)和有机污染物(BaP、BDE-47)对长牡蛎的复合毒性效应研究[D].北京:中国科学院大学, 2017:19Xie J. Combined toxic effects of heavy metals(Cd2+、Pb2+)and organic pollutants(BaP, BDE-47)on the oyster Crassostrea gigas[D]. Beijing:University of the Chinese Academy of Sciences, 2017:19(in Chinese)
[15] Syed S, Zubair A, Frieri M. Immune response to nanomaterials:Implications for medicine and literature review[J].Current Allergy&Asthma Reports, 2013, 13(1):50-57
[16] Gong N, Shao K S, Li G Y, et al. Nickel oxide nanoparticles induce oxidative stress and morphological changes onmarine Chlorella vulgaris[J]. Advanced Materials Research, 2014, 955-959(11):956-960
[17] Rikans L E, Hornbrook K R. Lipid peroxidation, antioxidant protection and aging[J]. Biochimica et Biophysica Acta, 1997, 1362(2-3):116-127
[18] Winston G W, Giulio R T. Prooxidant and antioxidant mechanisms in aquatic organisms[J]. Aquatic Toxicology,1991, 19(2):137-161
[19]陈剑杰,曹谨玲,李潇,等.氟对中华圆田螺肝胰脏抗氧化酶活性和MDA含量的影响[J].生态毒理学报,2018, 13(1):268-273Chen J J, Cao J L, Li X, et al. Effects of fluoride on the activities of antioxidant enzymes and MDA levels in hepatopancreas of Cipangopaludina cahayensis[J]. Asian Journal of Ecotoxicology, 2018, 13(1):268-273(in Chinese)
[20]马菲菲,孙雪梅,韩倩,等.不同粒径Ti O2颗粒对海洋微藻的毒性效应[J].海洋学报, 2015, 37(10):100-105Ma F F, Sun X M, Han Q, et al. Toxic effects of Ti O2particles with different size on the marine microalga[J]. Acta Oceanologica Sinica, 2015, 37(10):100-105(in Chinese)
[21] Slos S, Stoks R. Predation risk induces stress proteins and reduces antioxidant defense[J]. Functional Ecology, 2008,22(4):637-642
[22] Meng X L, Liu P, Li J, et al. Physiological responses of swimming crab Portunus trituberculatus under cold acclimation:Antioxidant defense and heat shock proteins[J].Aquaculture, 2014, 434:11-17
[23] Zalutskaya Z, Ostroukhova M, Filina V, et al. Nitric oxide upregulates expression of alternative oxidase 1 in Chlamydomonas reinhardtii[J]. Journal of Plant Physiology, 2017, 219:123-127
[24] Bhatia M. Understanding toxicology:Mechanisms and applications[J]. Cell Biology&Toxicology, 2017, 33(1):1-4
[25] Lachapelle G, Radicioni S M, Stankiewicz A R, et al. Acute acidification or amiloride treatment suppresses the ability of Hsp70 to inhibit heat-induced apoptosis[J]. Apoptosis, 2007, 12(8):1479-1488
[26] Ellison M A, Ferrier M D, Carney S L. Salinity stress results in differential Hsp70 expression in the Exaiptasia pallida and Symbiodinium symbiosis[J]. Marine Environmental Research, 2017, 132:63-67
[27] Mc Donald A E, Vanlerberghe G C, Staples J F. Alternative oxidase in animals:Unique characteristics and taxonomic distribution[J]. Journal of Experimental Biology,2009, 212(16):2627-2634
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