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硬壳蛤棕色细胞对镉急性反应中的体内金属硫蛋白和谷胱甘肽状态

In vivo metallothionein and glutathione status in an acute response to cadmium in Mercenaria mercenaria brown cells.

作者信息

Zaroogian G, Jackim E

机构信息

United States Environmental Protection Agency, Narragansett, RI 02882, USA.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2000 Dec;127(3):251-61. doi: 10.1016/s0742-8413(00)00152-3.

Abstract

Brown cells that are found in the red glands of Mercenaria mercenaria accumulate, detoxify and excrete cadmium. Brown cell involvement in metal detoxification was due in part to endogenous glutathione (GSH) and protein sulfhydryl. Metallothionein (MT) and GSH have been shown to play an important role in metal detoxification in bivalve molluscs. This study showed that the protein sulfhydryl in brown cells of Mercenaria was in fact MT, that brown cell GSH functioned in acute protection against Cd2+ toxicity, that GSH provided the initial defense against Cd2+ toxicity prior to MT induction and that MT variants were unequal in response to Cd2+. During treatment of Mercenaria with 0.5 and 1.0 ppm Cd2+, brown cells were analyzed for MT by capillary electrophoresis and GSH colorimetrically after 0.25, 1, 2, 3, and 4 days. The data indicated that the cadmium-binding protein was MT with an apparent molecular weight of 9 kDa determined by gel filtration or 6 kDa as indicated by capillary electrophoresis. Glutathione appeared to prevail in the brown cell acute response to 0.5 ppm Cd2+, whereas MT appeared to prevail in the acute response to 1.0 ppm Cd2+. Capillary electrophoresis can be used to monitor and quantify MT and its variants in brown cells without need for prior separation of cytosolic components by chromatography. The change in MT-II was greater relative to the change in MT-I in the brown cell acute response to 0.5 ppm Cd2+, whereas the change in MT-1 was greater relative to the change in MT-II in the acute response to 1.0 ppm Cd2+. The variants of brown cell MT appeared to respond differentially to Cd2+ depending upon the Cd2+ treatment concentration.

摘要

在硬壳蛤的红色腺体中发现的褐色细胞能够积累、解毒并排泄镉。褐色细胞参与金属解毒部分归因于内源性谷胱甘肽(GSH)和蛋白质巯基。金属硫蛋白(MT)和GSH已被证明在双壳贝类的金属解毒中发挥重要作用。本研究表明,硬壳蛤褐色细胞中的蛋白质巯基实际上就是MT,褐色细胞中的GSH在急性保护中发挥作用以抵御Cd2+毒性,在MT诱导之前GSH提供了对Cd2+毒性的初始防御,并且MT变体对Cd2+的反应并不相同。在用0.5 ppm和1.0 ppm Cd2+处理硬壳蛤的过程中,在0.25、1、2、3和4天后通过毛细管电泳分析褐色细胞中的MT,并通过比色法分析GSH。数据表明,镉结合蛋白是MT,通过凝胶过滤测定其表观分子量为9 kDa,而通过毛细管电泳测定为6 kDa。在褐色细胞对0.5 ppm Cd2+的急性反应中,谷胱甘肽似乎占主导地位,而在对1.0 ppm Cd2+的急性反应中,MT似乎占主导地位。毛细管电泳可用于监测和定量褐色细胞中的MT及其变体,无需事先通过色谱法分离细胞溶质成分。在褐色细胞对0.5 ppm Cd2+的急性反应中,MT-II的变化相对于MT-I的变化更大,而在对1.0 ppm Cd2+的急性反应中,MT-1的变化相对于MT-II的变化更大。褐色细胞MT的变体似乎根据Cd2+处理浓度对Cd2+有不同的反应。

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