Evaluation of the effect of silver and silver nanoparticles on the function of selenoproteins using an in-vitro model of the fish intestine: The cell line RTgutGC
Evaluation of the effect of silver and silver nanoparticles on the function of selenoproteins using an in-vitro model of the fish intestine: The cell line RTgutGC
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DOI：
10.1016/j.ecoenv.2021.111930
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发表时间：
2021-01-17
影响因子：
6.8
通讯作者：
Minghetti, Matteo
中科院分区：
文献类型：
--
作者：
Chanda, Debarati;Dudefoi, William;Minghetti, Matteo
在哺乳动物细胞中的新兴研究表明，离子（AgNO 3）和纳米银（AgNP）可以破坏硒的代谢，硒在氧化应激控制中起着至关重要的作用。然而，银（Ag）对鱼类硒蛋白功能的影响知之甚少。在这里，我们评估硝酸银和柠檬酸盐包被的AgNP（cit-AgNP）硒蛋白功能和氧化应激的影响，使用来自虹鳟鱼（虹鳟鱼）肠（RTgutGC）的鱼细胞系。使用同时测量代谢活性、膜完整性和溶酶体完整性的细胞毒性测定来评价细胞活力。暴露于等摩尔量的AgNO 3和cit-AgNP的细胞在细胞内积累相同量的银，然而AgNO 3比cit-AgNP毒性更大。硒酶谷胱甘肽过氧化物酶（GPx）和硫氧还蛋白还原酶（TrxR）的mRNA水平和酶活性进行了测定。虽然mRNA水平不受AgNO 3或cit-AgNP的影响，但GPx的酶活性受到AgNO 3（1 μ M）和cit-AgNP（5 μ M）的抑制，TrxR活性受到AgNO 3（0.4 μ M）和cit-AgNP（1.5 μ M）的抑制。此外，暴露于1 μ M AgNO 3和cit-AgNP的细胞在暴露24 h时显示金属硫蛋白B（MT B）mRNA水平增加，证实了银的摄取，但在72 h时恢复到对照水平，表明MT B清除了银。在所测试的AgNO 3或cit-AgNP的任何剂量下均未观察到氧化应激。总的来说，这项研究表明，AgNO 3或cit-AgNP可以抑制硒酶的活性，但不诱导RTgutGC细胞的氧化应激。
Emerging research in mammalian cells suggests that ionic (AgNO3) and nano silver (AgNP) can disrupt the metabolism of selenium which plays a vital role in oxidative stress control. However, the effect of silver (Ag) on selenoprotein function in fish is poorly understood. Here we evaluate the effects of AgNO3 and citrate coated AgNP (cit-AgNP) on selenoprotein function and oxidative stress using a fish cell line derived from the rainbow trout (Oncorhynchus mykiss) intestine (RTgutGC). Cell viability was evaluated using a cytotoxicity assay which measures simultaneously metabolic activity, membrane integrity and lysosome integrity. Cells exposed to equimolar amounts of AgNO3 and cit-AgNP accumulated the same amount of silver intracellularly, however AgNO3 was more toxic than cit-AgNP. Selenoenzymes glutathione peroxidase (GPx) and thioredoxin reductase (TrxR) mRNA levels and enzyme activity were measured. While mRNA levels remained unaffected by AgNO3 or cit-AgNP, the enzyme activity of GPx was inhibited by AgNO3 (1 mu M) and cit-AgNP (5 mu M) and TrxR activity was inhibited by AgNO3 (0.4 mu M) and cit-AgNP (1, 5 mu M). Moreover, cells exposed to 1 mu M of AgNO3 and cit-AgNP showed an increase in metallothionein b (MTb) mRNA levels at 24 h of exposure, confirming the uptake of silver, but returned to control levels at 72 h suggesting silver scavenging by MTb. Oxidative stress was not observed at any of the doses of AgNO3 or cit-AgNP tested. Overall, this study shows that AgNO3 or cit-AgNP can inhibit the activity of selenoenzymes but do not induce oxidative stress in RTgutGC cells.