背角无齿蚌超氧化物歧化酶基因的克隆及多溴联苯醚-47和多溴联苯醚-209对其表达的影响
Effects of PBDE-47 and PBDE-209 on the AwSOD Expression in Freshwater Bivalve Anodonta woodiana
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摘要: 超氧化物歧化酶（SOD）是机体防御和氧化应激损伤的关键酶之一。为了探讨多溴联苯醚PBDE-47和PBDE-209对背角无齿蚌的胁迫效应，本研究克隆出AwSOD全基因序列，分析PBDE-47和PBDE-209对AwSOD表达的影响。背角无齿蚌AwSOD cDNA全长由949个核苷酸组成，开放阅读框包含465 bp核苷酸，编码155个氨基酸。AwSOD包含Cu/ZnSOD家族GKHGFHVHEFGDNT和GNAGARSACGVI这2个特征性标签序列。AwSOD氨基酸序列与Cu/ZnSOD具有较高的同源性。AwSOD与淡水贝类亲缘关系最近，其次海洋双壳类，最后是脊椎动物、腹足类和甲壳类。AwSOD在肝胰脏中表达水平较高，在鳃和斧足为中等水平，在外套膜、闭壳肌和心脏中表达水平较低。与对照组相比，PBDE-47和PBDE-209处理对肝胰腺中AwSOD mRNA水平具有显著的诱导作用。PBDE-47处理对鳃中AwSOD mRNA水平在早期具有显著诱导作用，高剂量组在后期呈现下调效应；PBDE-209处理后鳃中AwSOD mRNA水平显著增加。结果表明，PBDE-47和PBDE-209对背角无齿蚌AwSOD表达水平诱导作用与增强机体抗氧化能力和胁迫效应有关。Abstract: The superoxide dismutases (SODs) play a key role against oxidative stress. In order to explore the stress of polybrominated diphenyl ethers (PBDEs) on the bivalve Anodonta woodiana, one complete Cu/ZnSOD sequence was isolated and named AwSOD in the current study. The full-length cDNA of AwSOD had an open reading frame (ORF) of 465 bp encoding 155 amino acids. The deduced amino acid sequence of AwSOD has high identity with the Cu/ZnSODs. The most relationship of evolution of AwSOD was that of freshwater bivalve, next was marine bivalve, last was vertebrate, gastropods and crustacean. In selected tissues, the highest expression of AwSOD mRNA was observed in the hepatopancreas, intermediate ones in the foot, adductor muscle, and gill, lower ones in heart and mantle. Compared with that of control group, AwSOD mRNA level of hepatopancreas was significantly induced by PBDE-47 and PBDE-209. AwSOD expression of gill was significantly up-regulated at early stage in the PBDE-47 treated groups, and down-regulated at later stage in the high-dose groups in contrasted with that of control group. AwSOD mRNA level of gill was significantly increased in the PBDE-209 treated goups. These results indicate that up-regulations of AwSOD expression of hepatopancreas and gill contributed to elimination of stress derived from PBDE-47 and PBDE-209 challenge in the freshwater bivalve A. woodiana.
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