双酚A暴露对不同黑色素含量长牡蛎的影响研究
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摘要
长牡蛎的壳色有很多种,其中壳色较深个体(黑壳)比较浅个体(白壳)的外套膜中含有更多的黑色素。本研究采用高效液相色谱-质谱法(HPLC-MS/MS)测定天然海水中白壳与黑壳长牡蛎外套膜双酚A (BPA)含量,发现黑壳长牡蛎外套膜中的BPA含量较白壳长牡蛎低(P<0.05)。由于黑色素合成过程中的关键——酶酪氨酸酶是一种酚氧化酶,可以通过对氧化酚类或芳胺类等多种底物的氧化起催化作用,推测其在BPA等酚类毒物的降解中也发挥重要作用。为了进一步研究不同黑色素含量牡蛎对BPA降解能力和对BPA损伤抵抗能力的差异,本实验进行了浓度为1 mg/L,1.5 mg/L和2 mg/L的BPA亚慢性暴露实验:通过对暴露后长牡蛎外套膜组织中BPA含量分析发现,白壳与黑壳长牡蛎外套膜中BPA含量均随着暴露浓度的升高而升高,且3种浓度BPA亚慢性暴露后,黑壳长牡蛎外套膜BPA含量均低于白壳长牡蛎,但是差异不显著(P>0.05)。采用3种浓度BPA亚慢性暴露后,黑壳长牡蛎外套膜组织中活性氧(ROS)荧光值均显著低于白壳长牡蛎(P<0.01)。通过对外套膜组织学观察发现,2 mg/L的高浓度BPA暴露使白壳长牡蛎外套膜组织发生了明显的病理变化,黑壳长牡蛎外套膜未发生明显的病理变化。可见,黑壳长牡蛎对BPA暴露可能具有更强的抵抗能力。
There are many variations in the shell color of Pacific oyster Crassostrea gigas, in which darker individuals(black shell) contains more melanin in the mantle than lighter individuals( white shell). In this study, high performance liquid chromatography-mass spectrometry(HPLC-MS/MS) was used to determine the content of bisphenol A(BPA) in the mantle of white shell and black shell oyster in natural seawater. It was found that the content of BPA in the mantle of black shell oyster was lower than that of white shell oyster(P<0.05). Since tyrosinase, a key enzyme in the synthesis of melanin, is a phenol oxidase, which can catalyze the oxidation of phenols or aromatic amines and other substrates, it is speculated that tyrosinase also plays an important role in the degradation of phenolic toxicants such as BPA. To further investigate the differences in the ability of BPA degradation and damage resistance between oysters with different melanin contents,subchronic exposure experiments of BPA with concentrations of 1 mg/L, 1.5 mg/L and 2 mg/L were performed in this study.Through the analysis of the BPA content in the mantle tissues of Crassostrea gigas after exposure, it was found that the BPA content in mantle of white and black shell oyster both increased with the rise of exposure concentration. In addition, after chronic exposure to BPA at the three concentrations, the BPA content in mantle of black shell oyster was lower than that of white shell oyster, but the difference was not significant(P>0.05). The fluorescence value of reactive oxygen species(ROS) in mantle of black shell oyster was significantly lower than that of white shell oyster after chronic exposure to BPA at three concentrations(P<0.01). The histological observation of the mantle revealed that BPA exposure caused obvious pathological changes in the mantle tissue of the white shell oyster, while there was no obvious pathological change in the mantle of the black shell oyster, suggesting that the black shell oyster may have stronger resistance to BPA exposure.
There are many variations in the shell color of Pacific oyster Crassostrea gigas, in which darker individuals(black shell) contains more melanin in the mantle than lighter individuals( white shell). In this study, high performance liquid chromatography-mass spectrometry(HPLC-MS/MS) was used to determine the content of bisphenol A(BPA) in the mantle of white shell and black shell oyster in natural seawater. It was found that the content of BPA in the mantle of black shell oyster was lower than that of white shell oyster(P<0.05). Since tyrosinase, a key enzyme in the synthesis of melanin, is a phenol oxidase, which can catalyze the oxidation of phenols or aromatic amines and other substrates, it is speculated that tyrosinase also plays an important role in the degradation of phenolic toxicants such as BPA. To further investigate the differences in the ability of BPA degradation and damage resistance between oysters with different melanin contents,subchronic exposure experiments of BPA with concentrations of 1 mg/L, 1.5 mg/L and 2 mg/L were performed in this study.Through the analysis of the BPA content in the mantle tissues of Crassostrea gigas after exposure, it was found that the BPA content in mantle of white and black shell oyster both increased with the rise of exposure concentration. In addition, after chronic exposure to BPA at the three concentrations, the BPA content in mantle of black shell oyster was lower than that of white shell oyster, but the difference was not significant(P>0.05). The fluorescence value of reactive oxygen species(ROS) in mantle of black shell oyster was significantly lower than that of white shell oyster after chronic exposure to BPA at three concentrations(P<0.01). The histological observation of the mantle revealed that BPA exposure caused obvious pathological changes in the mantle tissue of the white shell oyster, while there was no obvious pathological change in the mantle of the black shell oyster, suggesting that the black shell oyster may have stronger resistance to BPA exposure.
引文
Chen L,Xiang Y,Kong L,et al,2013.Hydroxysafflor yellow A protects against cerebral ischemia-reperfusion injury by anti-apoptotic effect through PI3K/Akt/GSK3βpathway in rat.Neurochemical Research38(11),2 268-2 275.
Crain D A,Eriksen M,Iguchi T,et al,2007.An ecological assessment of bisphenol-A:Evidence from comparative biology.Reproductive Toxicology 24(2),225-239.
Debecker S,Sommaruga R,Maes T,et al,2015.Larval UV exposure impairs adult immune function through a trade-off with larval investment in cuticular melanin.
Diamantikandarakis E,Bourguignon J P,Giudice L C,et al,2009.Endocrine-disrupting chemicals:an Endocrine Society scientific statement.Endocrine Reviews 30(4),293.
Evans S,Camara M D,Langdon C J,2009.Heritability of shell pigmentation in the Pacific oyster,Crassostrea gigas.Aquaculture286(3):211-216.
Flint S,Markle T,Thompson S,et al,2012.Bisphenol A exposure,effects,and policy:a wildlife perspective.Journal of Environmental Management 104(16):19-34.
Grégoire C,1967.On the structure of organic matrixes of molluscan shells.Biol Rev Camb Philos Soc 42(4):653-688.
Green T J,Dixon T J,Devic E,et al,2009.Differential expression of genes encoding anti-oxidant enzymes in Sydney rock oysters,Saccostrea glomerata(Gould)selected for disease resistance.Fish&Shellfish Immunology 26(5):799-810.
Hao S,Hou X,Wei L,et al,2016.Correction:Extraction and identification of the pigment in the adductor muscle scar of Pacific Oyster Crassostrea gigas.Plos One 11(2),e0148704.
Jin X,Jiang G,Huang G,et al,2004.Determination of 4-tertoctylphenol,4-nonylphenol and bisphenol A in surface waters from the Haihe River in Tianjin by gas chromatography-mass spectrometry with selected ion monitoring.Chemosphere 56(11):0-1119.
Jobling S,Casey D,Rodgers-Gray T,et al,2004.Comparative responses of molluscs and fish to environmental estrogens and an estrogenic effluent.Aquatic Toxicology 66(2):207-222.
Kaddar N,Bendridi N,HarthéC,et al,2009.Development of a radioimmunoassay for the measurement of Bisphenol A in biological samples.Analytica Chimica Acta 645(1):1-4.
Kang J H,Asai D,Katayama Y,2007.Bisphenol A in the aquatic environment and its endocrine-disruptive effects on aquatic organisms.Critical Reviews in Toxicology 37(7):607.
Kavlock R J,Daston G P,Derosa C,et al,1996.Research needs for the risk assessment of health and environmental effects of endocrine disruptors:a report of the U.S.EPA-sponsored workshop.Environ Health Perspect 104(Suppl 4):715-740.
Krishnan A V,Stathis P,Permuth S F,et al,1993.Bisphenol-A:an estrogenic substance is released from polycarbonate flasks during autoclaving.Endocrinology 132(6):2279.
Loschen G,Azzi A,Richter C,et al,1974.Superoxide radicals as precursors of mitochondrial hydrogen peroxide.Febs Letters 42(1):68-72.
Loschen G,FlohéL,Chance B,1971.Respiratory chain linked H2O2production in pigeon heart mitochondria.Febs Letters 18(2):261-264.
Luo L,Zhang Q,Kong X,et al,2017.Differential effects of bisphenol Atoxicity on oyster(Crassostrea angulata)gonads as revealed by labelfree quantitative proteomics.Chemosphere 176:305-314.
Mizuo K,Narita M,Miyagawa K,et al,2010.Effects of prenatal and neonatal exposure to bisphenol A on the development of the central nervous system Twenty-Seventh IEEE Applied Power Electronics Conference and Exposition,1368-1372.
MR K,XQ J H Y,2010.Innate immune responses of a lepidopteran insect,Manduca sexta.Immunological Reviews 198(1):97-105.
Naylor C G,Mieure J P,Adams W J,et al,1992.Alkylphenol ethoxylates in the environment.Journal of the American Oil Chemists Society 69(7):695-703.
Robinson C D,Brown E,Craft J A,et al,2003.Effects of sewage effluent and ethynyl oestradiol upon molecular markers of oestrogenic exposure,maturation and reproductive success in the sand goby(Pomatoschistus minutus,Pallas).Aquatic Toxicology 62(2):119-134.
Schulteoehlmann U,Tillmann M,Markert B,et al,2000.Effects of endocrine disruptors on prosobranch snails(mollusca:gastropoda)in the Laboratory.Part II:Triphenyltin as a Xeno-Androgen.Ecotoxicology 9(6):399-412.
Sohoni P,Tyler C R,Hurd K,et al,2001.Reproductive effects of longterm exposure to bisphenol A in the fathead minnow(Pimephales promelas).Environmental Science&Technology 35(14):2917-2925.
Staniszewska M,Koniecko I,Falkowska L,et al,2015.Occurrence and distribution of bisphenol A and alkylphenols in the water of the Gulf of Gdansk(Southern Baltic).Marine Pollution Bulletin 91(1):372-379.
Staples C A,Dome P B,Klecka G M,et al,1998.A review of the environmental fate,effects,and exposures of bisphenol A.Chemosphere 36(10):2149-2173.
Sweeney M F,Hasan N,Soto A M,et al,2015.Environmental endocrine disruptors:Effects on the human male reproductive system.Reviews in Endocrine&Metabolic Disorders 16(4):341-357.
Wu H,Zhong M,Lu Z,et al,2018.Biological effects of tris(1-chloro-2-propyl)phosphate(TCPP)on immunity in mussel Mytilus galloprovincialis.Environmental Toxicology&Pharmacology 61:102.
Zhang Y,Wang Q,Ji Y,et al,2014.Identification and mRNA expression of two 17β-hydroxysteroid dehydrogenase genes in the marine mussel Mytilus galloprovincialis following exposure to endocrine disrupting chemicals.Environmental Toxicology and Pharmacology 37(3):1243-1255.
陈清西,宋康康,2006.酪氨酸酶的研究进展.厦门大学学报(自然科学版),45(5):731-737.
邓旭修,黄会,徐英江,等,2014.渤海南部辛基酚和双酚A污染状况研究.海洋通报,33(2):222-227.
董军,李向丽,梁锐杰,2009.珠江口地区水体中双酚A污染及其与环境因子的关系.生态与农村环境学报,25(2):94-97.
何齐齐,宋丹,许雪峰,等,2018.三门湾海域污染物扩散数值研究.海洋通报.
黄少婵,杭义萍,2010.液相色谱-质谱法同时测定塑料制品中的双酚A和四溴双酚A.色谱,28(9):863-866.
黄辉洋,李少菁,王桂忠,2000.甲壳动物酚氧化酶活力及其在养殖中的应用.海洋通报,19(3):79-84.
蒋俊,李秀艳,2009.环境内分泌干扰物双酚A的降解研究进展.上海化工,34(4):25-30.
李正炎,傅明珠,王馨平,等,2006.冬季胶州湾及其周边河流中酚类环境激素的分布特征.中国海洋大学学报(自然科学版),36(3):113-117.
刘浩,戴明新,彭士涛,等,2011.渤海湾主要污染物环境容量的估算.海洋通报,30(4):451-455.
彭俊华,糜漫天,2004.环境雌激素辛基酚检测的研究进展.环境卫生学杂志,31(3):143-148.
孙胜龙,2005.环境激素与人类未来.北京:化学工业出版社,环境科学与工程出版中心.
孙卫玲,倪晋仁,郝鹏鹏,等,2004.泥沙对双酚A的吸附及其影响因素研究.环境科学学报,24(6):975-981.
王平,张宁川,2013.大连湾保守污染物迁移三维模型及应用.海洋通报,32(3):265-274.
肖国兵,袁伟,王仁元,等,2008.双酚A对接触男工精液质量影响的研究,宁波:宁波市学术大会.
张照韩,冯玉杰,高鹏,等,2011.松花江水内分泌干扰物及雌激素活性调查.哈尔滨工业大学学报,(12):58-62.
邹荣婕,邓旭修,王斌,等,2015.黄河调水调沙对黄河口海域双酚A的影响.生态学报,35(2):263-269.
Crain D A,Eriksen M,Iguchi T,et al,2007.An ecological assessment of bisphenol-A:Evidence from comparative biology.Reproductive Toxicology 24(2),225-239.
Debecker S,Sommaruga R,Maes T,et al,2015.Larval UV exposure impairs adult immune function through a trade-off with larval investment in cuticular melanin.
Diamantikandarakis E,Bourguignon J P,Giudice L C,et al,2009.Endocrine-disrupting chemicals:an Endocrine Society scientific statement.Endocrine Reviews 30(4),293.
Evans S,Camara M D,Langdon C J,2009.Heritability of shell pigmentation in the Pacific oyster,Crassostrea gigas.Aquaculture286(3):211-216.
Flint S,Markle T,Thompson S,et al,2012.Bisphenol A exposure,effects,and policy:a wildlife perspective.Journal of Environmental Management 104(16):19-34.
Grégoire C,1967.On the structure of organic matrixes of molluscan shells.Biol Rev Camb Philos Soc 42(4):653-688.
Green T J,Dixon T J,Devic E,et al,2009.Differential expression of genes encoding anti-oxidant enzymes in Sydney rock oysters,Saccostrea glomerata(Gould)selected for disease resistance.Fish&Shellfish Immunology 26(5):799-810.
Hao S,Hou X,Wei L,et al,2016.Correction:Extraction and identification of the pigment in the adductor muscle scar of Pacific Oyster Crassostrea gigas.Plos One 11(2),e0148704.
Jin X,Jiang G,Huang G,et al,2004.Determination of 4-tertoctylphenol,4-nonylphenol and bisphenol A in surface waters from the Haihe River in Tianjin by gas chromatography-mass spectrometry with selected ion monitoring.Chemosphere 56(11):0-1119.
Jobling S,Casey D,Rodgers-Gray T,et al,2004.Comparative responses of molluscs and fish to environmental estrogens and an estrogenic effluent.Aquatic Toxicology 66(2):207-222.
Kaddar N,Bendridi N,HarthéC,et al,2009.Development of a radioimmunoassay for the measurement of Bisphenol A in biological samples.Analytica Chimica Acta 645(1):1-4.
Kang J H,Asai D,Katayama Y,2007.Bisphenol A in the aquatic environment and its endocrine-disruptive effects on aquatic organisms.Critical Reviews in Toxicology 37(7):607.
Kavlock R J,Daston G P,Derosa C,et al,1996.Research needs for the risk assessment of health and environmental effects of endocrine disruptors:a report of the U.S.EPA-sponsored workshop.Environ Health Perspect 104(Suppl 4):715-740.
Krishnan A V,Stathis P,Permuth S F,et al,1993.Bisphenol-A:an estrogenic substance is released from polycarbonate flasks during autoclaving.Endocrinology 132(6):2279.
Loschen G,Azzi A,Richter C,et al,1974.Superoxide radicals as precursors of mitochondrial hydrogen peroxide.Febs Letters 42(1):68-72.
Loschen G,FlohéL,Chance B,1971.Respiratory chain linked H2O2production in pigeon heart mitochondria.Febs Letters 18(2):261-264.
Luo L,Zhang Q,Kong X,et al,2017.Differential effects of bisphenol Atoxicity on oyster(Crassostrea angulata)gonads as revealed by labelfree quantitative proteomics.Chemosphere 176:305-314.
Mizuo K,Narita M,Miyagawa K,et al,2010.Effects of prenatal and neonatal exposure to bisphenol A on the development of the central nervous system Twenty-Seventh IEEE Applied Power Electronics Conference and Exposition,1368-1372.
MR K,XQ J H Y,2010.Innate immune responses of a lepidopteran insect,Manduca sexta.Immunological Reviews 198(1):97-105.
Naylor C G,Mieure J P,Adams W J,et al,1992.Alkylphenol ethoxylates in the environment.Journal of the American Oil Chemists Society 69(7):695-703.
Robinson C D,Brown E,Craft J A,et al,2003.Effects of sewage effluent and ethynyl oestradiol upon molecular markers of oestrogenic exposure,maturation and reproductive success in the sand goby(Pomatoschistus minutus,Pallas).Aquatic Toxicology 62(2):119-134.
Schulteoehlmann U,Tillmann M,Markert B,et al,2000.Effects of endocrine disruptors on prosobranch snails(mollusca:gastropoda)in the Laboratory.Part II:Triphenyltin as a Xeno-Androgen.Ecotoxicology 9(6):399-412.
Sohoni P,Tyler C R,Hurd K,et al,2001.Reproductive effects of longterm exposure to bisphenol A in the fathead minnow(Pimephales promelas).Environmental Science&Technology 35(14):2917-2925.
Staniszewska M,Koniecko I,Falkowska L,et al,2015.Occurrence and distribution of bisphenol A and alkylphenols in the water of the Gulf of Gdansk(Southern Baltic).Marine Pollution Bulletin 91(1):372-379.
Staples C A,Dome P B,Klecka G M,et al,1998.A review of the environmental fate,effects,and exposures of bisphenol A.Chemosphere 36(10):2149-2173.
Sweeney M F,Hasan N,Soto A M,et al,2015.Environmental endocrine disruptors:Effects on the human male reproductive system.Reviews in Endocrine&Metabolic Disorders 16(4):341-357.
Wu H,Zhong M,Lu Z,et al,2018.Biological effects of tris(1-chloro-2-propyl)phosphate(TCPP)on immunity in mussel Mytilus galloprovincialis.Environmental Toxicology&Pharmacology 61:102.
Zhang Y,Wang Q,Ji Y,et al,2014.Identification and mRNA expression of two 17β-hydroxysteroid dehydrogenase genes in the marine mussel Mytilus galloprovincialis following exposure to endocrine disrupting chemicals.Environmental Toxicology and Pharmacology 37(3):1243-1255.
陈清西,宋康康,2006.酪氨酸酶的研究进展.厦门大学学报(自然科学版),45(5):731-737.
邓旭修,黄会,徐英江,等,2014.渤海南部辛基酚和双酚A污染状况研究.海洋通报,33(2):222-227.
董军,李向丽,梁锐杰,2009.珠江口地区水体中双酚A污染及其与环境因子的关系.生态与农村环境学报,25(2):94-97.
何齐齐,宋丹,许雪峰,等,2018.三门湾海域污染物扩散数值研究.海洋通报.
黄少婵,杭义萍,2010.液相色谱-质谱法同时测定塑料制品中的双酚A和四溴双酚A.色谱,28(9):863-866.
黄辉洋,李少菁,王桂忠,2000.甲壳动物酚氧化酶活力及其在养殖中的应用.海洋通报,19(3):79-84.
蒋俊,李秀艳,2009.环境内分泌干扰物双酚A的降解研究进展.上海化工,34(4):25-30.
李正炎,傅明珠,王馨平,等,2006.冬季胶州湾及其周边河流中酚类环境激素的分布特征.中国海洋大学学报(自然科学版),36(3):113-117.
刘浩,戴明新,彭士涛,等,2011.渤海湾主要污染物环境容量的估算.海洋通报,30(4):451-455.
彭俊华,糜漫天,2004.环境雌激素辛基酚检测的研究进展.环境卫生学杂志,31(3):143-148.
孙胜龙,2005.环境激素与人类未来.北京:化学工业出版社,环境科学与工程出版中心.
孙卫玲,倪晋仁,郝鹏鹏,等,2004.泥沙对双酚A的吸附及其影响因素研究.环境科学学报,24(6):975-981.
王平,张宁川,2013.大连湾保守污染物迁移三维模型及应用.海洋通报,32(3):265-274.
肖国兵,袁伟,王仁元,等,2008.双酚A对接触男工精液质量影响的研究,宁波:宁波市学术大会.
张照韩,冯玉杰,高鹏,等,2011.松花江水内分泌干扰物及雌激素活性调查.哈尔滨工业大学学报,(12):58-62.
邹荣婕,邓旭修,王斌,等,2015.黄河调水调沙对黄河口海域双酚A的影响.生态学报,35(2):263-269.