背角无齿蚌硒谷胱甘肽过氧化物酶基因的克隆及2,4-二氯苯酚、2,4,6-三氯苯酚和五氯苯酚对其表达的影响
Cloning of AwSeGPx from Anodonta woodiana and Effects of 2,4-dichlorophenol, 2,4,6-trichlorophenol and Pentachlorophenol on Its Transcription
-
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
-
-
Maheshwari N, Khan F H, Mahmood R. Pentachlorophenol-induced cytotoxicity in human erythrocytes:Enhanced generation of ROS and RNS, lowered antioxidant power, inhibition of glucose metabolism, and morphological changes[J]. Environmental Science and Pollution Research, 2019, 26(13):12985-13001 Matta R, Chiron S. Oxidative degradation of pentachlorophenol by permanganate for ISCO application[J]. Environmental Technology, 2018, 39(5):651-657 Yeo B J, Goh S, Zhang J S, et al. Novel MBRs for the removal of organic priority pollutants from industrial wastewaters:A review[J]. Journal of Chemical Technology & Biotechnology, 2015, 90(11):1949-1967 Lei B L, Huang S B, Jin X W, et al. Deriving the aquatic predicted no-effect concentrations (PNECs) of three chlorophenols for the Taihu Lake, China[J]. Journal of Environmental Science and Health, Part A, 2010, 45(14):1823-1831 Yin D Q, Gu Y, Li Y, et al. Pentachlorophenol treatment in vivo elevates point mutation rate in zebrafish p53 gene[J]. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 2006, 609(1):92-101 Maheshwari N, Mahmood R. 3,4-dihydroxybenzaldehyde attenuates pentachlorophenol-induced cytotoxicity, DNA damage and collapse of mitochondrial membrane potential in isolated human blood cells[J]. Drug and Chemical Toxicology, 2020, 28:1-18 Chatziargyriou V, Dailianis S. The role of selenium-dependent glutathione peroxidase (Se-GPx) against oxidative and genotoxic effects of mercury in haemocytes of mussel Mytilus galloprovincialis (Lmk.)[J]. Toxicology in Vitro, 2010, 24:1363-1372 Kim U J, Jo H, Lee I S, et al. Investigation of bioaccumulation and biotransformation of polybrominated diphenyl ethers, hydroxylated and methoxylated derivatives in varying trophic level freshwater fishes[J]. Chemosphere, 2015, 137:108-114 Soo P, Todd P A. The behaviour of giant clams (Bivalvia:Cardiidae:Tridacninae)[J]. Marine Biology, 2014, 161(12):2699-2717 夏西超, 张科, 宋国英, 等. 背角无齿蚌AwPrx4A基因克隆及在PFOS和PFOA胁迫下的表达分析[J]. 生态毒理学报, 2019, 14(6):118-127 Xia X C, Zhang K, Song G Y, et al. Effects of PFOS and PFOA on the AwPrx4A expression in freshwater bivalve Anodonta woodiana[J]. Asian Journal of Ecotoxicology, 2019, 14(6):118-127(in Chinese)
Xia X C, Xue S P, Wang X Y, et al. Response a chronic effects of PBDE-47:Up-regulations of HSP60 and HSP70 expression in freshwater bivalve Anodonta woodiana[J]. Fish & Shellfish Immunology, 2017, 65:213-225 Mullenbach G T, Tabrizi A, Irvine B D, et al. Sequence of a cDNA coding for human glutathione peroxidase confirms TGA encodes active site selenocysteine[J]. Nucleic Acids Research, 1987, 15(13):5484 De Zoysa M, Pushpamali W A, Oh C, et al. Transcriptional up-regulation of disk abalone selenium dependent glutathione peroxidase by H2O2 oxidative stress and Vibrio alginolyticus bacterial infection[J]. Fish & Shellfish Immunology, 2008, 25(4):446-457 Zhang L B, Liu X L, Chen L L, et al. Transcriptional regulation of selenium-dependent glutathione peroxidase from Venerupis philippinarum in response to pathogen and contaminants challenge[J]. Fish & Shellfish Immunology, 2011, 31(6):831-837 Hliwa P, Zdanowski B, Dietrich G J, et al. Temporal changes in gametogenesis of the invasive Chinese pond mussel Sinanodonta woodiana (Lea, 1834) (Bivalvia:Unionidae) from the Konin Lakes System (Central Poland)[J]. Folia Biologica, 2015, 63(3):175-185 Ladenstein R, Epp O, Gunzler W A, et al. Glutathione peroxidase on approval[J]. Life Chemistry Reports, 1986, 4(1):37-55 Aumann K D, Bedorf N, Brigelius-Flohed R, et al. Glutathione peroxidase revisited-Simulation of the catalytic cycle by computer-assisted molecular modelling[J]. Biomedical and Environmental Sciences, 1997, 10(Z1):136-155 Walczak R, Westhof E, Carbon P, et al. A novel RNA structural motif in the selenocysteine insertion element of eukaryotic selenoprotein mRNAs[J]. RNA, 1996, 2(4):367-379 Grundner-Culemann E, Martin G W, Harney J W, et al. Two distinct SECIS structures capable of directing selenocysteine incorporation in eukaryotes[J]. RNA, 1999, 5(5):625-635 Carneiro M, Reis B, Azevedo J, et al. Glutathione transferases responses induced by microcystin-LR in the gills and hepatopancreas of the clam Venerupis philippinarum[J]. Toxins, 2015, 7(6):2096-2120 Chen H M, Lee Y H, Wang Y J. ROS-triggered signaling pathways involved in the cytotoxicity and tumor promotion effects of pentachlorophenol and tetrachlorohydroquinone[J]. Chemical Research in Toxicology, 2015, 28(3):339-350 Pietsch C, Hollender J, Dorusch F, et al. Cytotoxic effects of pentachlorophenol (PCP) and its metabolite tetrachlorohydroquinone (TCHQ) on liver cells are modulated by antioxidants[J]. Cell Biology and Toxicology, 2014, 30(4):233-252 Doyen P, Rodius F, L'Hoste L, et al. Identification and mRNA expression of pi-class glutathione S-transferase and selenium-dependent glutathione peroxidase in the gudgeon Gobio gobio exposed to PCB 77[J]. Comparative Biochemistry and Physiology Part C:Toxicology & Pharmacology, 2012, 155(2):300-306 -
点击查看大图
计量
- 文章访问数: 1202
- HTML全文浏览数: 1202
- PDF下载数: 56
- 施引文献: 0
下载:
