袁凤娟,
魏梦薇,
张慧子,
王雯,
邵向阳,
李媛,
董艳美,
王梦琪,
齐金旭,
夏西超^,
平顶山学院医学院, 平顶山 476000

作者简介: 袁凤娟(1986-),女,硕士,讲师,研究方向为细胞增殖分化调控,E-mail:626189846@qq.com.

通讯作者: 夏西超,xiaxichao8336@163.com

基金项目: 河南省高等学校重点科研项目（20A330002）；河南省教育厅人文社会科学研究项目（2020-ZZJH-339）；平顶山学院博士启动基金资助项目（PXY-BSQD-2018009,PXY-PYJJ-2018005）


中图分类号: Q459

Cloning of AwSeGPx from Anodonta woodiana and Effects of 2,4-dichlorophenol, 2,4,6-trichlorophenol and Pentachlorophenol on Its Transcription

Yuan Fengjuan,
Wei Mengwei,
Zhang Huizi,
Wang Wen,
Shao Xiangyang,
Li Yuan,
Dong Yanmei,
Wang Mengqi,
Qi Jinxu,
Xia Xichao^,
College of Medicine, Pingdingshan University, Pingdingshan 467000, China

Corresponding author: Xia Xichao,xiaxichao8336@163.com

CLC number: Q459

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摘要:硒谷胱甘肽过氧化物酶（SeGPx）是一类广泛存在于生物体内的重要抗氧化酶，SeGPx可将有毒的过氧化物还原成无毒的羟基化合物，从而保护细胞膜结构及功能不受过氧化物干扰和损害。为了探讨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）对背角无齿蚌（Anodonta woodiana ）的胁迫效应，本研究克隆出AwSeGPx全基因序列，分析2，4-DCP、2，4，6-TCP和PCP对AwSeGPx表达的影响。AwSeGPx的cDNA序列全长870 bp，开放阅读框为585 bp，编码195个氨基酸。推导的AwSeGPx蛋白序列包括GPx家族标签序列（68LGFPCNQF75）和活性位点序列（156WNFEKF161）、一个典型终止密码子（165TGA167）编码的硒代半胱氨酸（U⁴⁴）、酶催化活性相关保守氨基酸位点：谷氨酰胺（Q⁷⁴）、精氨酸（R⁹⁴和R¹⁵¹）和色氨酸（W¹⁵⁶）。在相同浓度条件下，PCP对背角无齿蚌毒性效应大于2，4-DCP和2，4，6-TCP。与对照组相比，浓度60、120、240、480和960 μg·L^-1的2，4-DCP处理后肝胰腺AwSeGPx mRNA水平增加了1.79倍以上（P<0.05）；浓度100、200、400和800 μg·L^-1的2，4，6-TCP处理后AwSeGPx mRNA水平增加了1.01倍以上（P<0.05）；浓度20、40、80、160和320 μg·L^-1 PCP处理能够显著诱导AwSeGPx的表达。以上结果表明，2，4-DCP、2，4，6-TCP和PCP处理对背角无齿蚌肝胰腺中AwSeGPx表达具有明显的诱导作用，这种诱导效应与动物提高过氧化物的还原能力和增强环境的耐受能力密切相关。
关键词: 2,4-二氯苯酚/
2,4,6-三氯苯酚/
五氯苯酚/
背角无齿蚌/
AwSeGPx

Abstract:Selenium dependent glutathione peroxidase (SeGPx) belongs to the family of selenoprotein, which acts mainly as an antioxidant role in the cellular defense system. In order to explore the effect of 2,4-dichlorophenol (2,4-DCP), 2,4,6-trichlorophenol (2,4,6-TCP) and pentachlorophenol (PCP) on the freshwater bivalve Anodonta woodiana, a SeGPx full length cDNA was cloned from A. woodiana and named as AwSeGPx, and its expressions were analyzed by real-time PCR. 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 GPx active site (156WNFEKF160) were observed. It had a characteristic codon at 165TGA167 that corresponds to selenocysteine (Sec) amino acid as U⁴⁴. The catalytically related amino acid residues consisting of Gln (Q⁷⁴), Arg (R⁹⁴), Arg (R¹⁵¹) and Trp (W¹⁵⁶) were identified in the deduced AwSeGPx protein sequence. Under similar condition, clams A. woodiana showed more sensitive to PCP than 2,4-DCP and 2,4,6-TCP. In the 2,4-DCP treated groups (60, 120, 240, 480 and 960μ g·L^-1), AwSeGPx mRNA level of hepatopancreas increased more than 1.79 times (P<0.05) compared with that of control group. In the 2,4,6-TCP treated groups (100, 200, 400 and 800 μg·L^-1), AwSeGPx mRNA level of hepatopancreas increased more than 1.01 times (P<0.05). In the PCP treated groups (20, 40, 80, 160 and 320 μg·L^-1), expressions of AwSeGPx were significantly induced compared with that of control group in the hepatopancreas. These results indicate that AwSeGPx plays an active role in inhibiting oxidative stress derived from 2,4-DCP, 2,4,6-TCP and PCP treatment in the hepatopancreas.
Key words:2,4-dichlorophenol/
2,4,6-trichlorophenol/
pentachlorophenol/
Anodonta woodiana/
AwSeGPx.

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