摘要:
硫氧化蛋白过氧化物酶(Prx)可以将过氧化氢、有机过氧化物和过氧化合物分别转化为水、乙醇和亚硝酸盐,是机体一种重要的抗氧化蛋白。为了探讨全氟辛烷磺酸(PFOS)和全氟辛酸(PFOA)对背角无齿蚌的胁迫效应,背角无齿蚌随机分为对照组、PFOS处理组和PFOA处理组;同时克隆出AwPrx4A全基因序列,分析PFOS和PFOA对AwPrx4A表达的影响。背角无齿蚌AwPrx4A cDNA全长由958个核苷酸组成,包含1个120 bp的5’非编码区,1个412 bp的3’非编码区和1个426 bp的开放阅读框,开放阅读框为由142个氨基酸组成的多肽链。PFOS和PFOA对背角无齿蚌的LC50分别为28.388和192.083 mg·L-1。与对照组相比,浓度6.25、12.5、25、50和100 mg·L-1的PFOS处理后,实验观察过程中肝胰腺中AwPrx4A mRNA水平分别增加了18.75%、2.85倍(P<0.05)、5.08倍(P<0.01)、5.52倍(P<0.01)和6.77倍(P<0.01)以上。与对照组相比,浓度50、100、200、400和800 mg·L-1的PFOA处理后,实验观察过程中肝胰腺AwPrx4A mRNA水平分别增加了20.83%、2.21倍(P<0.01)、2.25倍(P<0.01)、3.19倍(P<0.01)和5.64倍(P<0.01)以上。与对照组相比,PFOS和PFOA处理后鳃中AwPrx4A mRNA水平分别增加了61.61%(P<0.05)和59.59%(P<0.05)以上。与对照组相比,PFOS和PFOA处理后血淋巴中AwPrx4A mRNA水平分别增加了47.42%和20.61%以上。结果表明,PFOS和PFOA处理对背角无齿蚌AwPrx4A表达具有明显的诱导作用,其原因与对抗PFOS和PFOA的胁迫效应有关。
Abstract:
Peroxiredoxin (Prx), a thiol specific antioxidant protein, plays an important role in eliminating reactive oxygen species (ROS). Perfluorooctane sulfonate (PFOS) and perfluoroocanoic acid (PFOA) are significant components of total perfluorinated chemicals and can catalyze the production of ROS in the body. In the current study, one complete Prx sequence was isolated from Anodonta woodiana and named AwPrx4A. The complete cDNA sequence of AwPrx4A was comprised of 958 nucleotides, including a 5’ untranslated region (UTR) of 120 bp, a long 3’ UTR of 412 bp, and a 426 bp open reading frame (ORF) which was encoded by 142 amino acids. Compared with that of control group, AwPrx4A mRNA level of hepatopancreas increased by more than 18.75%, 2.85 times (P<0.05), 5.08 times (P<0.01), 5.52 times (P<0.01) and 6.77 times (P<0.01) in the 6.25, 12.5, 25, 50 and 100 mg·L-1 of PFOS treated groups, respectively. Compared with that of control group, AwPrx4A mRNA level of hepatopancreas increased by more than 20.83%, 2.21 times (P<0.01), 2.25 times (P<0.01), 3.19 times (P<0.01) and 5.64 times (P<0.01) in the 50, 100, 200, 400 and 800 mg·L-1 of PFOA treated groups. Compared with that of control group, AwPrx4A mRNA level of gill increased by more than 61.61% (P<0.05) and 59.59% (P<0.05) in the PFOS treated groups and PFOA treated groups, respectively. Compared with that of control group, AwPrx4A mRNA level of hemocytes increased by more than 47.42% and 20.61% in the PFOS treated groups and PFOA treated groups, respectively. These results indicate that up-regulations of AwPrx4A expression of hepatopancreas, gill and hemocytes in the freshwater bivalve A. woodiana contribute to eliminating stress derived from PFOS and PFOA challenge.