单壁碳纳米管对太平洋牡蛎(Crassostrea gigas)的毒性效应及生物体防御机制研究
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摘要
碳纳米管的生态安全和健康风险日益受到人们的广泛关注。本文采用典型的海洋底栖生物——太平洋牡蛎(Crassostrea gigas, C. gigas)作为受试生物,研究了单壁碳纳米管(Single-Walled Carbon Nanotubes, SWCNTs)暴露对其造成的毒性效应及牡蛎自身的防御机制,以期为碳纳米管的海洋生态风险评价提供科学依据。在0.1～10 mg·L~(-1)的SWCNTs暴露96 h后,太平洋牡蛎鳃和消化腺中的丙二醛(malondialdehyde, MDA)含量显著增加(P≤0.05),总超氧化物歧化酶(superoxide dismutase, SOD)和过氧化氢酶(catalase, CAT)活性呈现显著的剂量依赖性升高(P≤0.05),cat、hsp70、aox及caspase-7等基因的相对表达量显著上调(P≤0.05)。相比于单独暴露,P-gp蛋白抑制剂Tariquidar与SWCNTs的复合暴露显著增加了鳃和消化腺中MDA含量,产生了更严重的氧化损伤。这些结果表明,SWCNTs暴露对太平洋牡蛎的鳃和消化腺造成了一定程度的氧化损伤,而牡蛎体内的抗氧化系统和多外源性物质抗性机制在防御SWCNTs的过程中起到了至关重要的作用。
The ecological and health risks of carbon nanotubes are receiving more attention. In this study, Pacific oyster(Crassostrea gigas), a kind of typical marine benthic organism, was exposed to single-walled carbon nanotubes(SWCNTs) to investigate the toxicity induced by SWCNTs and the defense mechanism of C.gigas. After 96-h exposure to 0.1-10 mg L-1 SWCNTs, the content of malondialdehyde(MDA) in gills and digestive glands increased significantly(P≤0.05), the activities of total superoxide dismutase(SOD) and catalase(CAT) showed the significant dose-dependent increase(P≤0.05), and the relative expressions of genes, i.e., cat, hsp70, aox and caspase-7, were significantly up-regulated(P≤0.05). The co-exposure of Tariquidar(P-gp protein inhibitor) and SWCNTs induced the significant increase of the MDA content in gills and digestive glands(P≤0.05), indicating that more serious oxidative damage were caused by the mixture. The results revealed that the antioxidant system and Multi-xenobiotic Resistance Mechanism played vital roles in the defense system of Pacific oyster against the SWCNTs exposure.
The ecological and health risks of carbon nanotubes are receiving more attention. In this study, Pacific oyster(Crassostrea gigas), a kind of typical marine benthic organism, was exposed to single-walled carbon nanotubes(SWCNTs) to investigate the toxicity induced by SWCNTs and the defense mechanism of C.gigas. After 96-h exposure to 0.1-10 mg L-1 SWCNTs, the content of malondialdehyde(MDA) in gills and digestive glands increased significantly(P≤0.05), the activities of total superoxide dismutase(SOD) and catalase(CAT) showed the significant dose-dependent increase(P≤0.05), and the relative expressions of genes, i.e., cat, hsp70, aox and caspase-7, were significantly up-regulated(P≤0.05). The co-exposure of Tariquidar(P-gp protein inhibitor) and SWCNTs induced the significant increase of the MDA content in gills and digestive glands(P≤0.05), indicating that more serious oxidative damage were caused by the mixture. The results revealed that the antioxidant system and Multi-xenobiotic Resistance Mechanism played vital roles in the defense system of Pacific oyster against the SWCNTs exposure.
引文
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[25]Zhou G,Kong Y,Bi Y,et al.Possible participation of active oxygen species in the induction of cyanide-resistant pathway at the initial senescence of tobacco callus[J].Russian Journal of Plant Physiology,2001,48(5):588-594
[26]Park C J,Seo Y S.Heat shock proteins:A review of the molecular chaperones for plant immunity[J].Plant Pathology Journal,2015,31(4):323-333
[27]Clegg J S,Uhlinger K R,Jackson S A,et al.Induced thermotolerance and the heat shock protein-70 family in the Pacific oyster Crassostrea gigas[J].Molecular Marine Biology and Biotechnology,1998,7(1):21-30
[28]Mukhopadhyay I,Nazir A,Saxena D K,et al.Heat shock response:Hsp70 in environmental monitoring[J].Journal of Biochemical&Molecular Toxicology,2003,17(5):249-254
[29]Hamdoun A M,Cheney D P,Cherr G N.Phenotypic plasticity of HSP70 and hsp70 gene expression in the Pacific oyster(Crassostrea gigas):Implications for thermal limits and induction of thermal tolerance[J].Biological Bulletin,2003,205(2):160-169
[30]Cicchetti R,Divizia M,Valentini F,et al.Effects of single-wall carbon nanotubes in human cells of the oral cavity:Geno-cytotoxic risk[J].Toxicology in Vitro,2011,25(8):1811-1819
[31]Epel D.Use of multidrug transporters as first lines of defense against toxins in aquatic organisms[J].Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,1998,120(1):23-28
[32]Ambudkar S V,Dey S,Hrycyna C A,et al.Biochemical,cellular,and pharmacological aspects of the multidrug transporter[J].Annual Review of Pharmacology and Toxicology,1999,39(1):361-398
[33]Huang L,Wang J,Chen W C,et al.P-glycoprotein expression in Perna viridis after exposure to Prorocentrum lima,a dinoflagellate producing DSP toxins[J].Fish&Shellfish Immunology,2014,39(2):254-262
[34]Palace V P,Evans R E,Wautier K G,et al.Interspecies differences in biochemical,histopathological,and population responses in four wild fish species exposed to ethynylestradiol added to a whole lake[J].Canadian Journal of Fisheries&Aquatic Sciences,2009,66(11):1920-1935
[2]Zhao F,Meng H,Yan L,et al.Nanosurface chemistry and dose govern the bioaccumulation and toxicity of carbon nanotubes,metal nanomaterials and quantum dots in vivo[J].Science Bulletin,2015,60(1):3-20
[3]Smith C J,Shaw B J,Handy R D.Toxicity of single walled carbon nanotubes to rainbow trout,(Oncorhynchus mykiss):Respiratory toxicity,organ pathologies,and other physiological effects[J].Aquatic Toxicology,2007,82(2):94-109
[4]Thakkar M,Mitra S,Wei L.Effect on growth,photosynthesis,and oxidative stress of single walled carbon nanotubes exposure to marine alga Dunaliella tertiolecta[J].Journal of Nanomaterials,2016(10):1-9
[5]Lovern S B,Klaper R.Daphnia magna mortality when exposed to titanium dioxide and fullerene(C60)nanoparticles[J].Environmental Toxicology&Chemistry,2010,25(4):1132-1137
[6]Zhang H,Slutsky A S,Vincent J L.Oxygen free radicals in ARDS,septic shock and organ dysfunction[J].Intensive Care Medicine,2000,26(4):474-476
[7]Sies H.Biological redox systems and oxidative stress[J].Cellular&Molecular Life Sciences,2007,64(17):2181-2188
[8]Canesi L,Ciacci C,Fabbri R,et al.Bivalve molluscs as a unique target group for nanoparticle toxicity[J].Marine Environmental Research,2012,76(4):16-21
[9]Ringwood A H,Levipolyachenko N,Carroll D L.Fullerene exposures with oysters:Embryonic,adult,and cellular responses[J].Environmental Science&Technology,2009,43(18):7136-7141
[10]Dagnino A,Allen J I,Moore M N,et al.Development of an expert system for the integration of biomarker responses in mussels into an animal health index[J].Biomarkers,2007,12(2):155-172
[11]Poland C A,Duffin R,Kinloch I,et al.Carbon nanotubes introduced into the abdominal cavity of mice show asbestos-like pathogenicity in a pilot study[J].Nature Nanotechnology,2008,3(7):423-428
[12]Ringwood A H,Conners D E,Dinovo A.The effects of copper exposures on cellular responses in oysters[J].Marine Environmental Research,1998,46(1-5):591-595
[13]Trevisan R,Delapedra G,Mello D F,et al.Gills are an initial target of zinc oxide nanoparticles in oysters Crassostrea gigas,leading to mitochondrial disruption and oxidative stress[J].Aquatic Toxicology,2014,153(8):27-38
[14]D'Agata A,Fasulo S,Dallas L J,et al.Enhanced toxicity of'bulk'titanium dioxide compared to'fresh'and'aged'nano-Ti O2in marine mussels(Mytilus galloprovincialis)[J].Nanotoxicology,2014,8(5):549-558
[15]Hughes F M,Foster B,Grewal S,et al.Apoptosis as a host defense mechanism in Crassostrea virginica and its modulation by Perkinsus marinus[J].Fish&Shellfish Immunology,2010,29(2):247-257
[16]Wang X.The expanding role of mitochondria in apoptosis[J].Genes&Development,2001,15(22):2922-2933
[17]葛春梅,黄茜枝,林道辉,等.人工纳米材料对贝类生态毒理效应的研究进展[J].生态毒理学报,2015,10(4):1-16Ge C M,Huang X Z,Wang Y J,et al.Research progress in ecotoxic effects of manufactured nanomaterials on shellfish[J].Asian Journal of Ecotoxicology,2015,10(4):1-16(in Chinese)
[18]Gong N,Shao K S,Li G Y,et al.Nickel oxide nanoparticles induce oxidative stress and morphological changes on marine Chlorella vulgaris[J].Advanced Materials Research,2014,955-959(11):956-960
[19]Tedesco S,Doyle H,Redmond G,et al.Gold nanoparticles and oxidative stress in Mytilus edulis[J].Marine Environmental Research,2008,66(1):131-133
[20]Canesi L,Fabbri R,Gallo G,et al.Biomarkers in Mytilus galloprovincialis exposed to suspensions of selected nanoparticles(Nano carbon black,C60fullerene,Nano-Ti O2,Nano-Si O2)[J].Aquatic Toxicology,2010,100(2):168-177
[21]Pan J F,Buffet P E,Poirier L,et al.Size dependent bioaccumulation and ecotoxicity of gold nanoparticles in an endobenthic invertebrate:The Tellinid clam Scrobicularia plana[J].Environmental Pollution,2012,168(3):37-43
[22]Yip J Y,Vanlerberghe G C.Mitochondrial alternative oxidase acts to dampen the generation of active oxygen species during a period of rapid respiration induced to support a high rate of nutrient uptake[J].Physiologia Plantarum,2010,112(3):327-333
[23]Purvis A C,Shewfelt R L.Does the alternative pathway ameliorate chilling injury in sensitive plant tissues?[J].Physiologia Plantarum,1993,88(4):712-718
[24]Maxwell D P,Wang Y,Mcintosh L.The alternative oxidase lowers mitochondrial reactive oxygen production in plant cells[J].Proceedings of the National Academy of Sciences of the United States of America,1999,96(14):8271-8276
[25]Zhou G,Kong Y,Bi Y,et al.Possible participation of active oxygen species in the induction of cyanide-resistant pathway at the initial senescence of tobacco callus[J].Russian Journal of Plant Physiology,2001,48(5):588-594
[26]Park C J,Seo Y S.Heat shock proteins:A review of the molecular chaperones for plant immunity[J].Plant Pathology Journal,2015,31(4):323-333
[27]Clegg J S,Uhlinger K R,Jackson S A,et al.Induced thermotolerance and the heat shock protein-70 family in the Pacific oyster Crassostrea gigas[J].Molecular Marine Biology and Biotechnology,1998,7(1):21-30
[28]Mukhopadhyay I,Nazir A,Saxena D K,et al.Heat shock response:Hsp70 in environmental monitoring[J].Journal of Biochemical&Molecular Toxicology,2003,17(5):249-254
[29]Hamdoun A M,Cheney D P,Cherr G N.Phenotypic plasticity of HSP70 and hsp70 gene expression in the Pacific oyster(Crassostrea gigas):Implications for thermal limits and induction of thermal tolerance[J].Biological Bulletin,2003,205(2):160-169
[30]Cicchetti R,Divizia M,Valentini F,et al.Effects of single-wall carbon nanotubes in human cells of the oral cavity:Geno-cytotoxic risk[J].Toxicology in Vitro,2011,25(8):1811-1819
[31]Epel D.Use of multidrug transporters as first lines of defense against toxins in aquatic organisms[J].Comparative Biochemistry and Physiology Part A:Molecular&Integrative Physiology,1998,120(1):23-28
[32]Ambudkar S V,Dey S,Hrycyna C A,et al.Biochemical,cellular,and pharmacological aspects of the multidrug transporter[J].Annual Review of Pharmacology and Toxicology,1999,39(1):361-398
[33]Huang L,Wang J,Chen W C,et al.P-glycoprotein expression in Perna viridis after exposure to Prorocentrum lima,a dinoflagellate producing DSP toxins[J].Fish&Shellfish Immunology,2014,39(2):254-262
[34]Palace V P,Evans R E,Wautier K G,et al.Interspecies differences in biochemical,histopathological,and population responses in four wild fish species exposed to ethynylestradiol added to a whole lake[J].Canadian Journal of Fisheries&Aquatic Sciences,2009,66(11):1920-1935