虾夷扇贝内脏团对镉的富集特性及生理响应
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摘要
【目的】对虾夷扇贝(Mizuhopectenyessoensis)内脏团中镉的富集特性进行研究,并分析镉暴露后与镉胁迫相关的基因在内脏团中的表达。【方法】采用人为添加镉的方法使虾夷扇贝进行镉暴露,原子吸收分光光度计测定各组织中镉含量,实时荧光定量PCR技术检测过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPx)和热休克蛋白70(HSP70)基因在虾夷扇贝内脏团中的表达。【结果】虾夷扇贝内脏团比其他组织富集了更多的镉(P=0.004),在含400μg/L Cd海水中暴露7 d和10 d后,扇贝内脏团中的Cd质量分数由62μg/g(干基)分别增加到180和217μg/g(干基);内脏团中抗氧化酶CAT、SOD和GPx以及热休克蛋白HSP70在200μg/L Cd暴露7 d后的基因表达量也显著增加(P <0.05)。【结论】内脏团是虾夷扇贝富集镉的重要组织,内脏团中CAT、SOD、GPx和HSP70对扇贝镉的防御有重要作用。
【Objective】To study the cadmium accumulation characteristics of Mizuhopecten yessoensis and to investigate the expression of several genes related to cadmium stress in the digestive gland of scallop after cadmium exposure. 【Method】 Scallops were exposed to cadmium by artificial addition,the cadmium contents in different tissues were determined by atomic absorption spectrophotometer, the expressions of CAT, SOD, GPx and HSP70 genes in the digestive gland of scallop were detected by quantitative real-time PCR. 【Result】Comparing to other tissues, higher concentration of cadmium was accumulated in the digestive gland of scallop(P = 0.004). After exposure to seawater containing 400μg/L of cadmium for 7 days and 10 days, the concentration of Cd in the digestive gland of scallop increased from 62 μg/g(dry weight) to 180 and 217 μg/g(dry weight), respectively. Gene expression of antioxidant enzymes CAT, SOD, GPx and heat shock protein HSP70 in the digestive gland were also increased significantly after the scallop was exposed to 200 μg/L of cadmium for 7 days(P < 0.05).【Conclusion】The digestive gland is an important organ for cadmium accumulation in M. yessoensis.The CAT, SOD, GPx and HSP70 genes play important roles in the defense cadmium stress of scallop.
【Objective】To study the cadmium accumulation characteristics of Mizuhopecten yessoensis and to investigate the expression of several genes related to cadmium stress in the digestive gland of scallop after cadmium exposure. 【Method】 Scallops were exposed to cadmium by artificial addition,the cadmium contents in different tissues were determined by atomic absorption spectrophotometer, the expressions of CAT, SOD, GPx and HSP70 genes in the digestive gland of scallop were detected by quantitative real-time PCR. 【Result】Comparing to other tissues, higher concentration of cadmium was accumulated in the digestive gland of scallop(P = 0.004). After exposure to seawater containing 400μg/L of cadmium for 7 days and 10 days, the concentration of Cd in the digestive gland of scallop increased from 62 μg/g(dry weight) to 180 and 217 μg/g(dry weight), respectively. Gene expression of antioxidant enzymes CAT, SOD, GPx and heat shock protein HSP70 in the digestive gland were also increased significantly after the scallop was exposed to 200 μg/L of cadmium for 7 days(P < 0.05).【Conclusion】The digestive gland is an important organ for cadmium accumulation in M. yessoensis.The CAT, SOD, GPx and HSP70 genes play important roles in the defense cadmium stress of scallop.
引文
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[20]FANG Y,YANG H,WANG T,et al.Metallothionein and superoxide dismutase responses to sublethal cadmium exposure in the clam Mactra veneriformis[J].Comp Biochem Physiol C,2010,151(3):325-333.
[21]MARKLUND S L,WESTMAN N G,LUNDGREN E,et al.Copper-and zinc-containing superoxide dismutase,manganese-containing superoxide dismutase,catalase,and glutathione peroxidase in normal and neoplastic human cell lines and normal human tissues[J].Cancer Res,1982,42(5):1955-1961.
[22]KARRAY S,MARCHAND J,MOREAU B,et al.Transcriptional response of stress-regulated genes to cadmium exposure in the cockle Cerastoderma glaucum from the gulf of Gabès area(Tunisia)[J].Environ Sci Pollut Res,2015,22(22):17290-17302.
[23]REGOLI F,NIGRO M,BERTOLI E,et al.Defenses against oxidative stress in the Antarctic scallop Adamussium colbecki and effects of acute exposure to metals[J].Hydrobiologia,1997,355(1/2/3):139-144.
[24]TLILI S,MINGUEZ L,GIAMBERINI L,et al.Assessment of the health status of Donax trunculus from the Gulf of Tunis using integrative biomarker indices[J].Ecol Indic,2013,32:285-293.
[25]陈晓聪,张冉,李成华,等.菲律宾蛤仔(Venerupis philippinarum)对重金属Hg2+的富集及相关生物标记物的识别[J].海洋与湖沼,2015,46(4):918-936.
[2]SERAFIM A,COSSON R,CAMUS L,et al.The effect of cadmium on antioxidant responses and the susceptibility to oxidative stress in the hydrothermal vent mussel Bathymodiolus azoricus[J].Mar Biol,2006,148(4):817-825.
[3]WATANABE M,HENMI,K,OGAWA K.et al.Cadmium-dependent generation of reactive oxygen species and mitochondrial DNA breaks in photosynthetic and non-photosynthetic strains of Euglena gracilis[J].Comp Biochem Physiol C,2003,134(2):227-234.
[4]WINSTON G W.Oxidants and antioxidants in aquatic animals[J].Comp Biochem Physiol,1991,100(1/2):173-176.
[5]高翔,王源.贝类镉胁迫功能基因研究进展[J].中国农业大学学报,2017,22(3):122-130.
[6]LOAIZA I,HURTADO D,MIGLIO M,et al.Tissue-specific Cd and Pb accumulation in Peruvian scallop(Argopecten purpuratus)transplanted to a suspended and bottom culture at Sechura Bay,Peru[J].Marine Pollution Bulletin,2015,91(2):429-440.
[7]LI C,NI D,SONG L,et al.Molecular cloning and characterization of a catalase gene from Zhikong scallop Chlamys farreri[J].Fish Shellfish Immun,2008,24(1):26-34.
[8]FANG Y,YANG H,LIU B,et al.Transcriptional response of lysozyme,metallothionein,and superoxide dismutase to combined exposure to heavy metals and bacteria in Mactra veneriformis[J].Comp Biochem Physiol C,2013,157(1):54-62.
[9]REGOLI F,PRINCIPATO G.Glutathione,glutathionedependent and antioxidant enzymes in mussel,Mytilus galloprovincialis,exposed to metals under field and laboratory conditions:implications for the use of biochemical biomarkers[J].Aquat Toxicol,1995,31(2):143-164.
[10]刘慧慧,李云娟,何建瑜,等.热休克及菌胁迫对厚壳贻贝HSP70基因表达的影响[J].水产学报,2015,39(7):971-977
[11]EVTUSHENKO Z S,Lukyanova O N,BELCHEVA N N.Cadmium bioaccumulation in organs of the scallop Mizuhopecten yessoensis[J].Mar Biol,1990,104(2):247-250.
[12]GAO J,ISHIZAKI S,NAGASHIMA Y.Purification and characterization of metal-binding proteins from the digestive gland of the Japanese scallop Mizuhopecten yessoensis[J].Fisheries Science,2016,82(2):337-345.
[13]高加龙,章超桦,梁家辉,等.湿法消化-石墨炉原子吸收法测定贝肉中镉含量的条件优化[J].现代食品科技,2012,28(11):1585-1589.
[14]陈海刚,林钦,蔡文贵,等.3种常见海洋贝类对重金属Hg Pb和Cd的积累与释放特征比较[J].农业环境科学学报,2008,27(3):1163-1167.
[15]WANG J,ZHANG H,ZHANG T.et al.Molecular mechanism on cadmium-induced activity changes of catalase and superoxide dismutase[J].Int J Biol Macromol,2015,77:59-67.
[16]MOORE M N,ALLEN J I.A computational model of the digestive gland epithelial cell of marine mussels and its simulated responses to oil-derived aromatic hydrocarbons[J].Mar Environ Res,2002,54(3/4/5):579-584.
[17]JO P G,CHOI Y K,CHOI C Y,2008.Cloning and m RNA expression of antioxidant enzymes in the Pacific oyster,Crassostrea gigas in response to cadmium exposure[J].Comp Biochem Physiol C,147(4):460-469.
[18]WANG Q,WANG X,WANG X,et al.Analysis of metallotionein expression and antioxidant enzyme activities in Meretrix meretrix larvae under sublethal cadmium exposure[J].Aquat Toxicol,2010,100(4):321-328.
[19]NI D,SONG L,GAO Q,et al.The c DNA cloning and m RNA expression of cytoplasmic Cu,Zn superoxide dismutase(SOD)gene in scallop Chlamys farreri[J].Fish Shellfish Immun,2007,23(5):1032-1042.
[20]FANG Y,YANG H,WANG T,et al.Metallothionein and superoxide dismutase responses to sublethal cadmium exposure in the clam Mactra veneriformis[J].Comp Biochem Physiol C,2010,151(3):325-333.
[21]MARKLUND S L,WESTMAN N G,LUNDGREN E,et al.Copper-and zinc-containing superoxide dismutase,manganese-containing superoxide dismutase,catalase,and glutathione peroxidase in normal and neoplastic human cell lines and normal human tissues[J].Cancer Res,1982,42(5):1955-1961.
[22]KARRAY S,MARCHAND J,MOREAU B,et al.Transcriptional response of stress-regulated genes to cadmium exposure in the cockle Cerastoderma glaucum from the gulf of Gabès area(Tunisia)[J].Environ Sci Pollut Res,2015,22(22):17290-17302.
[23]REGOLI F,NIGRO M,BERTOLI E,et al.Defenses against oxidative stress in the Antarctic scallop Adamussium colbecki and effects of acute exposure to metals[J].Hydrobiologia,1997,355(1/2/3):139-144.
[24]TLILI S,MINGUEZ L,GIAMBERINI L,et al.Assessment of the health status of Donax trunculus from the Gulf of Tunis using integrative biomarker indices[J].Ecol Indic,2013,32:285-293.
[25]陈晓聪,张冉,李成华,等.菲律宾蛤仔(Venerupis philippinarum)对重金属Hg2+的富集及相关生物标记物的识别[J].海洋与湖沼,2015,46(4):918-936.