1,4二恶烷对鱼体淋巴细胞的损伤效应研究

摘要1,4二恶烷是一类新型持久性有机污染物，具有环境持久性、生物富集性以及生物毒性效应，有关1,4二恶烷环境污染行为的研究已成为近年来环境化学的热点。1,4二恶烷作为一种新型持久性环境污染物，会对哺乳动物体内的抗氧化系统造成影响，进而对哺乳动物的生存造成影响。本研究选用藻食性鲤科鲫鱼（Carassius auratus）为实验材料，1,4二恶烷为目标污染物，采用离体细胞体外诱导和暴露培养方法，通过MTT细胞增殖法及细胞毒性检测法对细胞生长抑制率进行测定，并且利用酶标仪以及分光光度计对鲫鱼淋巴细胞的抗氧化酶系统指标进行测定。我们将鲫鱼解剖后提取出其肾脏，再从中提取出淋巴细胞，并将其暴露在0.1微克/升、1微克/升、5微克/升、10微克/升、100微克/升、500微克/升、1000微克/升、10000微克/升的1,4二恶烷中12小时，最后测定淋巴细胞的生长抑制率(MTT)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽转移酶(GST)和谷胱甘肽还原酶(GR)，进一步说明1,4二恶烷造成水生生物氧化损伤的影响机制。实验结果显示，随着1,4二恶烷暴露浓度的上升，SOD、CAT和GR的活性降低，GST活性增加，细胞生长抑制率逐渐增加，并呈现显著的剂量-效应关系。由此表明，高浓度的1,4二恶烷可诱导鲫鱼淋巴细胞抗氧化酶系统活性减小，造成淋巴细胞内产生氧化应激反应，脂质发生过氧化，进一步导致细胞损伤甚至凋亡。52719

毕业论文关键词：1,4二恶烷；水生生物；淋巴细胞；抗氧化酶系统；

Abstract Studies on 1,4-dioxane have become a hot topic in environmental chemistry region, because it’s a new type of persistent organic pollutants, which shows environmental persistence, bioaccumulation and toxicity effects. 1,4-dioxane affects antioxidant defense system and viability of mammals. This study selected lymphocytes from kidneys of algal feeding cyprinid fish (Carassius auratus) as experimental materials,. using 1,4-dioxane as the target pollutant. The cells inhibition percentages were measured by MTT proliferation, cytotoxicity assay and index of oxidative stress using an ELIASA and UV spectrophotometry. Lymphocytes were exposed to 0.1μg/L, 1μg/L, 5μg/L, 10μg/L, 100μg/L, 500μg/L, 1000μg/ L and 10000μg/L 1,4-dioxane for 12 hours. Then the cell growth inhibition percentage activity of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR) and glutathione transferase (GST) were determined to explain the mechanism of 1,4-dioxane -induced oxidative damage to aquatic organisms. Our results showed that with the increasing concentration of 1,4-dioxane, the activities of SOD, CAT and GR decreased, while the activity of GST and cell growth inhibition percentage increased gradually with dose-dependent manners. All results showed that high concentration of 1,4-dioxane exposure can induce cytotoxicity of Carassius auratus lymphocytes, such as oxidative stress, lipid peroxidation, even apoptosis.

Keyword: 1,4-dioxane; aquatic organisms; lymphocytes; antioxidant system;

摘要 3

Abstract 4

目录 5

1 前言 6

2 材料及方法 8

2.1 试验鱼种 8

2.2 毒素和试剂 8

2.3 淋巴细胞分离与诱导 8

2.4 MTT细胞增殖检测 8

2.5 超氧化物歧化酶（SOD）WST-1法测定 8

2.6 过氧化氢酶(CAT)活性测定 1,4二恶烷对鱼体淋巴细胞的损伤效应研究:http://www.youerw.com/yixue/lunwen_56715.html