养殖水体邻苯二甲酸二乙酯暴露对凡纳滨对虾壳膜几丁质酶的影响
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摘要
邻苯二甲酸二乙酯(DEP)是常用的塑化剂,也是环境广泛存在的内分泌干扰物。采用酶促反应动力学法及实时荧光定量PCR检测分析养殖水体DEP暴露对凡纳滨对虾(Litopenaeus vannamei)壳膜几丁质酶的比活力和酶系列基因mRNA表达量的影响。结果显示不同浓度组的DEP暴露对壳膜几丁质酶比活力的影响不具有显著性;10μg/L DEP暴露120 h后,对虾壳膜几丁质酶系的LV-CHT2、LV-CHT3、LV-CHT7等三个基因的mRNA表达量均被显著抑制,但随着暴露时间延长,这种显著性差异转化成非显著性;而20μg/L DEP胁迫对LV-CHT2、LV-CHT3、LV-CHT7等三个基因的mRNA表达量没有显著性影响,不同浓度组的DEP暴露不同时间后对LVCHT5、LV-CHT6等两个基因的mRNA表达量也均不具有显著性影响。
Diethyl phthalate(DEP) is one of the most common plasticizers. It is also a widely spreaded environmental endocrine disruptor. In this study,the effects of DEP exposure on the activity and mRNA expressions of Litopenaeus vannamei epidermal chitinase were investigated by using enzymatic reaction dynamic method and the real-time fluorescent quantitative PCR. The results showed that DEP exposure had no significant influence on the activity of epidermal chitinase. The mRNA expressions of epidermal chitinase gene such as LV-CHT2,LV-CHT3,LV-CHT7 were significantly inhibited after exposure to 10 μg/L DEP for 120 h. However,data significance disappeared after the exposures were continued. The 20 μg/L DEP had no significant effects on the mRNA expression of LV-CHT2,LV-CHT3 and LV-CHT7. Similar effects were observed for the mRNA expressions of LV-CHT5 and LV-CHT6 after DEP exposure.
Diethyl phthalate(DEP) is one of the most common plasticizers. It is also a widely spreaded environmental endocrine disruptor. In this study,the effects of DEP exposure on the activity and mRNA expressions of Litopenaeus vannamei epidermal chitinase were investigated by using enzymatic reaction dynamic method and the real-time fluorescent quantitative PCR. The results showed that DEP exposure had no significant influence on the activity of epidermal chitinase. The mRNA expressions of epidermal chitinase gene such as LV-CHT2,LV-CHT3,LV-CHT7 were significantly inhibited after exposure to 10 μg/L DEP for 120 h. However,data significance disappeared after the exposures were continued. The 20 μg/L DEP had no significant effects on the mRNA expression of LV-CHT2,LV-CHT3 and LV-CHT7. Similar effects were observed for the mRNA expressions of LV-CHT5 and LV-CHT6 after DEP exposure.
引文
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[2]葛健,林芳,韩宝瑜.邻苯二甲酸二乙酯(DEP)在罗非鱼(Oreochromis niloticus×O.aureus)体内代谢动力学和残留研究[J].海洋与湖沼,2011,42(1):137-141.
[3]Wang J L,Ye Y C,Wu W Z.Comparison of di-n-meth ylphthalate biodegradation by free and immobilized microbial cells[J].Biomed Environ Sci,2003,16(2):126-132.
[4]陆洋,袁东星,邓永智.九龙江水源水及其出厂水邻苯二甲酸酯污染调查[J].环境与健康杂志,2007,24(9):730-735.
[5]庄婉娥,姚文松,汪厦霞,等.泉州湾海水及沉积物中PAEs的分布及化学组成[J].环境与健康杂志,2011,28(10):898-902.
[6]姜琳琳.闽南地区水产品中邻苯二甲酸酯类(PAEs)的分布特征及风险评价[J].渔业科学进展,2015,36(4):139-144.
[7]王兴强,曹梅,阎斌伦,等.塑料增塑剂对凡纳滨对虾存活、生长和免疫的影响[J].生态毒理学报,2010,5(1):69-75.
[8]杜娟,谢晓兰,石艳,等.患红体病的凡纳滨对虾外壳膜NAGase基本性质的改变[J].应用与环境生物学报,2008,14(2):207-210.
[9]Huang Q S,Yan J H,Tang J Y,et al.Cloning and tissue expressions of seven chitinase family genes in Litopenaeus vannamei[J].Fish Shellfish Immunol,2010,29(1):75-81.
[10]余瑞元,袁明秀,陈丽蓉,等.生物化学实验原理和方法[M].第二版.北京:北京大学出版社,2005:240-242.
[11]谢晓兰,曾炜,高平章,等.黄粉虫几丁质酶的纯化及性质[J].泉州师范学院学报,2012,30(4):56-61.
[12]Livak K J,Schmittgen T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCTmethod[J].Methods,2002,25(4):402-408.
[13]李海涛,黄岁樑.水环境中邻苯二甲酸酯的迁移转化研究[J].环境污染与防治,2005,28(11):127-135.
[14]Zhao Z Y,Yin Z X,Weng S P,et al.Profiling of differentially expressed genes in hepatopancreas of white spot syndrome virusresistant shrimp(Litopenaeus vannamei)by suppression subtractive hybridization[J].Fish Shellfish Immunol,2007,22(5):520-534.