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亚砷酸盐对虹鳟鱼中肠 GST 和 ABCC2 活性和表达的离体和体内影响。

Ex vivo and in vivo effects of arsenite on GST and ABCC2 activity and expression in the middle intestine of the rainbow trout Oncorhynchus mykiss.

机构信息

Laboratorio de Ecotoxicología Acuática, INIBIOMA (CONICET-UNCo) - CEAN, ruta provincial no. 61, km 3, CCP 7, Junín de los Andes, 8371 Neuquén, Argentina.

Instituto de Biotecnologia Agropecuaria del Comahue, Sub-Sede Ibac, CITAAC (CONICET-UNCo) Universidad Nacional del Comahue, Buenos Aires 1400. Neuquén 8300, Argentina.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2019 Nov;225:108566. doi: 10.1016/j.cbpc.2019.108566. Epub 2019 Jul 10.

Abstract

In fish of freshwaters environments, the accumulation and toxic effects of arsenite (AsIII) can be attenuated by detoxification proteins such as GST and ABCC transporters. We studied the effects of AsIII on the middle intestine of O. mykiss in ex-vivo and in vivo/ex vivo assays. For the ex vivo assays, we measured the transport rate of the ABCC substrate DNP-SG and GST activity in intestinal strips and everted sacs. AsIII inhibited DNP-SG transport in a concentration-dependent manner, specifically when we applied it on the basolateral side. GST activity increased when we applied a maximum concentration of AsIII. For the in vivo/ex vivo assays, we kept fish in water with or without 7.7 μmol L of AsIII for 48 h. Then, we measured DNP-SG transport rate, GST activity, and PP1 activity in intestine strips during one hour. For PP1 activity, we incubated the strips with or without microcystin-LR (MCLR), a toxin excreted through ABCC2 proteins. We also analyzed Abcc2 and Gst-π mRNA expression in intestine and liver tissue. In the group exposed in vivo to AsIII, DNP-SG transport rate and GST activity were higher and the effect of MCLR over PP1 activity was attenuated. AsIII significantly induced only Abcc2 mRNA expression in both middle intestine and liver. Our results suggest that, in the middle intestine of O. mykiss, AsIII is absorbed mainly at the basolateral side of the enterocytes, excreted to the lumen by ABCC2 transporters, and is capable of modulating Abcc2 mRNA expression by a transcriptional mechanism.

摘要

在淡水环境中的鱼类中,谷胱甘肽 S-转移酶 (GST) 和 ABCC 转运体等解毒蛋白可以减轻亚砷酸盐 (AsIII) 的积累和毒性作用。我们研究了 AsIII 对 O. mykiss 中肠的体外和体内/体外测定的影响。对于体外测定,我们测量了 ABCC 底物 DNP-SG 的转运率和肠条和外翻囊中的 GST 活性。AsIII 以浓度依赖的方式抑制 DNP-SG 转运,特别是当我们将其施加到基底外侧时。当我们施加最大浓度的 AsIII 时,GST 活性增加。对于体内/体外测定,我们将鱼置于含有或不含有 7.7 μmol L 的 AsIII 的水中 48 小时。然后,我们在一小时内测量肠条中的 DNP-SG 转运率、GST 活性和 PP1 活性。对于 PP1 活性,我们用或不用微囊藻毒素-LR (MCLR) 孵育条带,MCLR 是通过 ABCC2 蛋白排泄的毒素。我们还分析了肠和肝组织中 Abcc2 和 Gst-π mRNA 的表达。在体内暴露于 AsIII 的组中,DNP-SG 转运率和 GST 活性较高,并且 MCLR 对 PP1 活性的影响减弱。AsIII 仅在中肠和肝组织中显著诱导 Abcc2 mRNA 表达。我们的结果表明,在 O. mykiss 的中肠中,AsIII 主要在肠细胞的基底外侧被吸收,通过 ABCC2 转运体排泄到腔中,并且能够通过转录机制调节 Abcc2 mRNA 表达。

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