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摘要全氟辛酸（PFOA）是全氟类有机化合物（PFCs）的一种，是一种新型的持久性有机污染物，具有极强的化学稳定性和生物蓄积性，已在全球范围内广泛存在，对生物安全性构成威胁，成为全球性问题。本文旨在研究PFOA对鲫鱼淋巴细胞凋亡效应的诱导机制，从而表明其对鲫鱼的免疫毒性以及致毒机理。我们将鲫鱼解剖后提取出头肾，再从中提取出淋巴细胞，将其暴露在1mg/L、5mg/L、15mg/L和30mg/L的PFOA中12小时，最后测定淋巴细胞的凋亡率、DNA-Ladder、丙二醛MDA、活性氧簇ROS以及谷胱甘肽GSH。结果显示，低剂量的PFOA的暴露就可引起ROS水平和MDA含量的显著升高（P<0.05）与GSH含量的极显著下降（P<0.01)，表明氧化应激反应的发生，使脂质发生过氧化；同时细胞凋亡因子的测定结果与DNA-Ladder条带也显示出细胞凋亡率上升。此外，随着PFOA暴露浓度的上升，MDA含量和ROS水平都随之上升，差异极为显著（P<0.01）并表现出剂量-效应关系，与此同时，GSH含量也随之下降，具有剂量-效应关系并且差异极为显著（P<0.01）。随着脂质过氧化的发生，细胞凋亡率与DNA-Ladder也表现出细胞进一步凋亡并且凋亡率上升。以上结果表明：PFOA能够诱导鲫鱼淋巴细胞发生氧化应激反应，导致脂质过氧化，进一步发生细胞凋亡，从而对鲫鱼的免疫系统产生危害。58286
Abstract PFOA, one of the perfluorinated compounds（PFCs）,is a new-typed persistent organic pollutant with strong chemical stability. It is able to accumulate the in the organisms and has been discharged in the most parts of the world. Consequently, it has been detected worldwide and its pollution has been determined as a global environmental problem. The study aims to research on the apoptosis of lymphocyte on Carassius auratus by Perfluorooctanoate (PFOA) in vitro, and so as to demonstrate PFOA’s immunotoxicity on crucian and its toxic mechanics. The crucians have been dissected and then the head-kidney has been taken out which is used to obtain lymphocyte. The lymphocyte samples have been pided to 5 groups. One of them has been set as controlled group and the others have been exposed to PFOA 1mg/L, 5mg/L, 15mg/L and 30mg/L for 12 hours. In the end, we analyzed the percent of apoptotic cells and DNA-Ladder to observe the apoptosis effects; furthermore, the antioxidant enzyme levels including MDA, GSH and ROS have been tested too. The results show that the low-dose PFOA can cause the striking increase of ROS level and MDA content (P<0.05) and the significant decrease of GSH content. It shows that the cells are enduring the oxidative stress and lipid peroxidation happened. Simultaneously, the results from DNA-Ladder and apoptotic cells test show the percent of apoptotic cells increased. Moreover, with the increase of PFOA exposed dose, MDA content and ROS level are increasing continuously with significance (P<0.01), and the increasing tendency shows the dose-effect condition. Meanwhile, GSH content decreases constantly and significantly (P<0.01), which also shows the dose-effect condition. In addition, with the phenomenon of lipid peroxidation, the results of apoptotic cell test and DNA-Ladder also show the apoptosis phenomenon becomes more serious. In summary, PFOA exposing is able to cause lipid peroxidation and so as to induce the apoptosis of lymphocyte on Carassius auratus, which damages the immunity of Carassius auratus.
毕业论文关键词：全氟辛酸； 鲫鱼淋巴细胞； 毒效应；氧化应激
Keyword: PFOA; crucian lymphocyte; toxicity; oxidative Stress
目 录
1 引言 4
2 材料与方法 6
2.1 试验鱼种 6
2.2 试剂和毒素 6
2.3 淋巴细胞分离与诱导