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流式细胞术在分析罗非鱼血液参数中的应用:在环境水生态毒理学中的应用。

Flow cytometry in the analysis of hematological parameters of tilapias: applications in environmental aquatic toxicology.

机构信息

Department of Morphology, UFMG, Belo Horizonte, MG, Brazil.

Department of Pharmacology, UFMG, Belo Horizonte, MG, Brazil.

出版信息

Environ Sci Pollut Res Int. 2021 Feb;28(5):6242-6248. doi: 10.1007/s11356-020-12119-7. Epub 2021 Jan 4.

Abstract

Blood tissue has been used to assess animal health and the environment in which they live. This tissue is easily acquired and has the ability to respond to various adverse conditions. Several techniques have been employed in the detection of xenobiotic-induced cell damage in blood cells. In general, traditionally used technologies, such as cellular analysis in blood smears, are time-consuming and require great analytical capacity. The present study proposes flow cytometry as a method to detect changes in blood cell populations. Tilapia (Oreochromis niloticus) was selected as a model for plotting the profile of fish blood cell populations after exposure to xenobiotics without euthanizing animals or using cell markers. Populations of erythrocytes and lymphocytes were detected only by combining the techniques of FACSAria cell sorting and light microscopy. Systemic deleterious effects were found through blood analysis, such as an increased lymphocyte-rich population at 48 h of exposure followed by a subsequent decrease. Moreover, the time-dependent expression of Nrf2 suggests its participation in increased membrane disruption, indicating it has a central role in erythrocyte lifespan. The present results shed light on the viability of using flow cytometry for blood analysis of living fish.

摘要

血液组织已被用于评估动物的健康状况及其生活环境。这种组织很容易获取,并且能够对各种不利条件做出反应。已经采用了几种技术来检测血液细胞中外来物质诱导的细胞损伤。一般来说,传统的技术,如血液涂片的细胞分析,既耗时又需要大量的分析能力。本研究提出流式细胞术作为一种检测血细胞群变化的方法。选用罗非鱼(Oreochromis niloticus)作为模型,在不处死动物或使用细胞标记物的情况下,研究暴露于外来物质后鱼类血细胞群的分布情况。通过结合 FACSAria 细胞分选和光学显微镜技术,仅检测到红细胞和淋巴细胞的群体。通过血液分析发现了全身性的有害影响,例如暴露 48 小时后淋巴细胞丰富的群体增加,随后减少。此外,Nrf2 的时间依赖性表达表明其参与了膜破裂的增加,表明其在红细胞寿命中起核心作用。本研究结果表明,使用流式细胞术对活体鱼类进行血液分析是可行的。

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