CdSe 纳米晶体对海洋硅藻三角褐指藻的毒理学效应:基于质谱的蛋白质组学的首次应用。

Toxicological effects of CdSe nanocrystals on the marine diatom Phaeodactylum tricornutum: The first mass spectrometry-based proteomic approach.

机构信息

Institut National des Sciences et Techniques de la Mer, Conservatoire National des Arts et Métiers, 50103 Cherbourg en Cotentin Cedex, France; Laboratoire Universitaire des Sciences Appliquées de Cherbourg, EA4253, Normandie Université, UNICAEN, 50130 Cherbourg en Cotentin, France.

Institut National des Sciences et Techniques de la Mer, Conservatoire National des Arts et Métiers, 50103 Cherbourg en Cotentin Cedex, France; IFREMER, LEAD NC, Equipe Ecophysiologie Station aquacole de Saint Vincent, Boulouparis, 98897 Nouvelle Calédonie Cedex, France.

出版信息

Ecotoxicol Environ Saf. 2018 May 15;152:78-90. doi: 10.1016/j.ecoenv.2018.01.043. Epub 2018 Feb 4.

Abstract

UNLABELLED

In the marine environment, benthic diatoms from estuarine and coastal sediments are among the first targets of nanoparticle pollution whose potential toxicity on marine organisms is still largely unknown. It is therefore relevant to improve our knowledge of interactions between these new pollutants and microalgae, the key players in the control of marine resources. In this study, the response of P. tricornutum to CdSe nanocrystals (CdSe NPs) of 5 nm (NP5) and 12 nm (NP12) in diameter was evaluated through microscopic, physiological, biochemical and proteomic approaches. NP5 and NP12 affected cell growth but oxygen production was only slightly decreased by NP5 after 1-d incubation time. In our experimental conditions, a high CdSe NP dissolution was observed during the first day of culture, leading to Cd bioaccumulation and oxidative stress, particularly with NP12. However, after a 7-day incubation time, proteomic analysis highlighted that P. tricornutum responded to CdSe NP toxicity by regulating numerous proteins involved in protection against oxidative stress, cellular redox homeostasis, Ca regulation and signalling, S-nitrosylation and S-glutathionylation processes and cell damage repair. These proteome changes allowed algae cells to regulate their intracellular ROS level in contaminated cultures. P. tricornutum was also capable to control its intracellular Cd concentration at a sufficiently low level to preserve its growth. To our knowledge, this is the first work allowing the identification of proteins differentially expressed by P. tricornutum subjected to NPs and thus the understanding of some molecular pathways involved in its cellular response to nanoparticles.

SIGNIFICANCE

The microalgae play a key role in the control of marine resources. Moreover, they produce 50% of the atmospheric oxygen. CdSe NPs are extensively used in the industry of renewable energies and it is regrettably expected that these pollutants will sometime soon appear in the marine environment through surface runoff, urban effluents and rivers. Since estuarine and coastal sediments concentrate pollutants, benthic microalgae which live in superficial sediments will be among the first targets of nanoparticle pollution. Thus, it is relevant to improve our knowledge of interactions between diatoms and nanoparticles. Proteomics is a powerful tool for understanding the molecular mechanisms triggered by nanoparticle exposure, and our study is the first one to use this tool to identify proteins differentially expressed by P. tricornutum subjected to CdSe nanocrystals. This work is fundamental to improve our knowledge about the defence mechanisms developed by algae cells to counteract damage caused by CdSe NPs.

摘要

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在海洋环境中,来自河口和沿海沉积物的底栖硅藻是纳米颗粒污染的首批目标之一,其对海洋生物的潜在毒性在很大程度上尚不清楚。因此,提高我们对这些新污染物与微藻(控制海洋资源的关键参与者)之间相互作用的认识是很重要的。在这项研究中,通过微观、生理、生化和蛋白质组学方法评估了 P. tricornutum 对直径为 5nm(NP5)和 12nm(NP12)的 CdSe 纳米晶体(CdSe NPs)的反应。NP5 和 NP12 影响细胞生长,但 NP5 在 1 天孵育时间后仅略微降低了产氧量。在我们的实验条件下,在培养的第一天观察到 CdSe NP 高度溶解,导致 Cd 生物积累和氧化应激,特别是用 NP12 时。然而,在 7 天的孵育时间后,蛋白质组分析强调 P. tricornutum 通过调节许多参与抗氧化应激、细胞氧化还原稳态、Ca 调节和信号转导、S-亚硝基化和 S-谷胱甘肽化过程以及细胞损伤修复的蛋白质来响应 CdSe NP 毒性。这些蛋白质组变化使藻类细胞能够在受污染的培养物中调节其细胞内 ROS 水平。P. tricornutum 还能够将其细胞内的 Cd 浓度控制在足够低的水平以维持其生长。据我们所知,这是首次通过 P. tricornutum 鉴定出受 NPs 影响的差异表达蛋白的工作,从而理解其细胞对纳米颗粒反应的一些分子途径。

意义

微藻在控制海洋资源方面发挥着关键作用。此外,它们产生了 50%的大气氧气。CdSe NPs 在可再生能源工业中被广泛使用,令人遗憾的是,这些污染物将很快通过地表径流、城市废水和河流出现在海洋环境中。由于河口和沿海沉积物浓缩了污染物,生活在表层沉积物中的底栖微藻将是首批受到纳米颗粒污染的目标之一。因此,提高我们对硅藻和纳米颗粒之间相互作用的认识是很重要的。蛋白质组学是理解纳米颗粒暴露引发的分子机制的有力工具,我们的研究首次使用该工具来鉴定 P. tricornutum 受 CdSe 纳米晶体影响的差异表达蛋白。这项工作对于提高我们对藻类细胞为抵抗 CdSe NPs 造成的损害而开发的防御机制的认识是基础性的。

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