不同结构全氟化合物对鱼体免疫细胞氧化损伤效应的比较研究

摘要全氟化合物（PFCs）是一类常见的有机化合物，它主要包括有全氟辛酸（PFOA）、全氟十二烷酸（PFDoA）和全氟十四烷酸（PFTA）等，该类物质的安全性受到了国内外研究工作者极大的重视。本文旨在研究PFOA、PFDoA、PFTA分别对免疫细胞的氧化损伤效应，比较三者之间的损伤效应差异。本研究选用藻食性鲤科鲫鱼（Carassius auratus）为实验材料，通过解剖，提取肾组织的淋巴细胞，将其暴露分别1ug/L、10ug/L、100ug/L、1000ug/L和10000ug/L的浓度PFOA、PFDoA、PFTA中12h。通过测定暴露后细胞的活性氧（ROS）、丙二醛(MDA)、微量还原型谷胱甘肽（GSH）、超氧化物歧化酶（SOD）等指标研究这三者对鲫鱼免疫毒性的机理，推测结构对全氟化合物的免疫毒性的影响。结果显示低浓度PFOA、PFDoA、PFTA的暴露均会引起ROS水平和MDA含量的显著上升，GSH含量极显著下降（P<0.01），SOD的活力下降趋势明显其抑制率显著上升（P<0.01），但三者同浓度暴露时，其对应指标变化相差较大。以上结果均表明：PFOA、PFDoA、PFTA均能够抑制SOD细胞抗氧化作用并且诱导鲫鱼淋巴细胞积累大量的ROS使细胞发生氧化应激，引起脂质过氧化产生MDA，导致GSH水平下降造成机体氧化应激和抗氧化能力衰减，进一步发生细胞氧化损伤，从而对鲫鱼的免疫系统产生危害。61363

Abstract Perfluorinated compounds (PFCs) is a class of common organic compounds, such as perfluorooctanoic acid (PFOA), security perfluorinated dodecanoic acid (PFDoA) and tetradecanoic acid (PFTA), etc. This paper aims to study the effects of PFOS, PFoA, PFCA on immune cells of carp, and compare the difference of three perfluorinated compounds. In this study, algae-feeding carp (Carassius auratus) was selected as experiment materials. After extracting lymphocytes from carp renal tissue by dissecting, cells were exposed to 1μg/L, 10μg/L, 100μg/L, 1000μg/L and 10000μg/L PFOA, PFDoA, PFTA for 12h. Reactive oxygen species (ROS), malondialdehyde (MDA), trace glutathione (GSH), superoxide dismutase (SOD) were detected to illustrate immunotoxicity mechanism and demonstrate the immunotoxicity related to the structure of perfluorinated compounds. Results showed that low exposure concentrations of PFOA, PFDoA, PFTA can cause significant increase in ROS level and MDA content, but significant decrease of GSH contents (p<0.01) and SOD activities (p<0.01). However, compared with three different structures exposed to the same concentration, the corresponding indexes changes were quite different. These results showed that PFOA, PFDoA, PFTA can induce oxidative damage in lymphocytes and affect the immune system by inhibiting the SOD antioxidant effects, increasing the ROS levels and declining GSH levels.

毕业论文关键词：全氟辛酸；全氟十二酸；全氟十四酸；氧化损伤；免疫细胞

Keyword: perfluorooctanoic acid; perfluorinated dodecanoic acid; tetradecanoic acid; oxidative damage；immune Cells

1前言 1

2 材料及方法 3

2.1 试验鱼种 3

2.2 毒素和试剂 4

2.3 淋巴细胞分离与诱导 4

2.4 MTT法细胞增殖及细胞毒性检测 4

2.6 流式细胞仪检测细胞内活性氧（ROS）检测 5

2.7 微量丙二醛(MDA)检测 5

2.8 谷胱甘肽(GSH)的测定 5

2.9超氧化物歧化酶（SOD）检测 6

3 结果 6

3.1 PFOA、PFDoA和PFTA诱导对鲫鱼淋巴细胞的ROS含量影响不同结构全氟化合物对鱼体免疫细胞氧化损伤效应的比较研究:http://www.youerw.com/shengwu/lunwen_67108.html