Xia Xichao, Hua Chunxiu, Xue Shipeng, Shi Bingqin, Gui Gaixia, Zhang Dongxian, Wang Xiying, Guo Lianghong
Basal Medicine Institution of Nanyang Medical College, Nanyang 473041, Henan Province, China; State Key Laboratory of Environmental Chemistry and Eco-toxicology, Research Centre for Eco-environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.
Basal Medicine Institution of Nanyang Medical College, Nanyang 473041, Henan Province, China.
Fish Shellfish Immunol. 2016 Aug;55:499-509. doi: 10.1016/j.fsi.2016.06.012. Epub 2016 Jun 9.
2,4-dichlorophenol (2,4-DCP), 2,4,6-trichlorophenol (2,4,6-TCP), and pentachlorophenol (PCP) pose a health risk to aquatic organism and humans, and are recognized as persistent priority pollutants. Selenium dependent glutathione peroxidase (Se-GPx) belongs to the family of selenoprotein, which acts mainly as an antioxidant role in the cellular defense system. In the current study, a Se-GPx full length cDNA was cloned from Anodonta woodiana and named as AwSeGPx. It had a characteristic codon at 165TGA167 that corresponds to selenocysteine(Sec) amino acid as U44. The full length cDNA consists of 870 bp, an open reading frame (ORF) of 585 bp encoded a polypeptide of 195 amino in which conserved domain (68LGFPCNQF75) and a glutathione peroxide-1 GPx active site (32GKVILVENVASLUGTT47) were observed. Additionally, the eukaryotic selenocysteine insertion sequence (SECIS) was conserved in the 3'UTR. The AwSeGPx amino acid sequence exhibited a high similarity with that of other Se-GPx. Real-time PCR analysis revealed that AwSeGPx mRNA had a widely distribution, but the highest level was observed in hepatopancreas. AwSeGPx mRNA expression was significantly up-regulated in hepatopancreas, gill and hemocytes after 2,4-DCP, 2,4,6-TCP and PCP exposure. Under similar environment, clams A. woodiana showed a more sensitive to PCP than that of 2,4-DCP and 2,4,6-TCP. These results indicate that AwSeGPx plays a protective role in eliminating oxidative stress derived from 2,4-DCP, 2,4,6-TCP and PCP treatment.
2,4-二氯苯酚(2,4-DCP)、2,4,6-三氯苯酚(2,4,6-TCP)和五氯苯酚(PCP)对水生生物和人类健康构成风险,被公认为持久性优先污染物。硒依赖性谷胱甘肽过氧化物酶(Se-GPx)属于硒蛋白家族,在细胞防御系统中主要起抗氧化作用。在本研究中,从背角无齿蚌克隆到一个Se-GPx全长cDNA,命名为AwSeGPx。它在165TGA167处有一个特征密码子,对应于作为U44的硒代半胱氨酸(Sec)氨基酸。全长cDNA由870 bp组成,一个585 bp的开放阅读框(ORF)编码一个195个氨基酸的多肽,其中观察到保守结构域(68LGFPCNQF75)和一个谷胱甘肽过氧化物-1 GPx活性位点(32GKVILVENVASLUGTT47)。此外,真核硒代半胱氨酸插入序列(SECIS)在3'UTR中保守。AwSeGPx氨基酸序列与其他Se-GPx的氨基酸序列具有高度相似性。实时PCR分析表明,AwSeGPx mRNA分布广泛,但在肝胰腺中水平最高。在2,4-DCP、2,4,6-TCP和PCP暴露后,肝胰腺、鳃和血细胞中AwSeGPx mRNA表达显著上调。在相似环境下,背角无齿蚌对PCP的敏感性高于2,4-DCP和2,4,6-TCP。这些结果表明,AwSeGPx在消除2,4-DCP、2,4,6-TCP和PCP处理产生的氧化应激中起保护作用。