二苯甲酮对斑点叉尾鮰鱼卵巢细胞生长的影响-环境科学专业论文.docxVIP

摘 要 二苯甲酮（Benzophenone）作为一种紫外线吸收剂，广泛应用于化工、医药、 建材等行业已有三十多年的历史，本文研究二苯甲酮对叉尾鮰鱼卵巢细胞的毒性。 实验首先利用斑点叉尾鮰鱼卵巢细胞（CCO）、鲤鱼上皮癌细胞（EPC）、草鱼肾细 胞（CIK）探讨经改良后的四噻唑蓝（MTT）比色法在测定污染物对鱼类细胞急性毒 性实验中的最佳条件及其使用的普遍性。而后在上述条件下，通过直接、间接两种 染毒条件来探讨二苯甲酮（benzophenone）对斑点叉尾鮰鱼卵巢细胞（CCO）生长的 影响，结果得到： 1、MTT 比色法应用于鱼类细胞时，甲瓒溶解液可以选取 14%的 SDS-DMF （pH=4.7）酸性溶液，并且采用保留培养液直接投加溶解液的办法溶解甲瓒，经讨 论这种方法在鱼类体外培养的细胞系的毒性试验中具有一定的普遍性。其最佳实验 条件为 570nm 波长下，MTT 作用 4 小时，MTT 终浓度 1mg/ml，最佳接种量的确定 因每种细胞的特性而不同，生长速度快的细胞接种量较少。 2、当二苯甲酮以 1μg/mL~10μg/mL 的浓度直接作用于 CCO 细胞时，CCO 细胞 生长在不同阶段均受到抑制作用：在指数增长期细胞生长速率减小，稳定期的存活 量约为正常条件下的 72—91%，并且随着二苯甲酮浓度的增大，对 CCO 生长的抑制 效应也相应的加剧。 3、当 1μg/mL 浓度的二苯甲酮在经过斑点叉尾鮰鱼原代肝细胞代谢后间接作用 于 CCO 细胞时，其代谢产物对 CCO 的生长在各阶段又呈现明显的促进作用，稳定 期细胞数量约为正常条件下的 109%。 关键词：改良 MTT 比色法，二苯甲酮，斑点叉尾鮰鱼卵巢细胞，生长，影响 Abstract Benzophenone is an UV-absorbing agent that has been used in industry, material and medicine for more than 30 years. In order to determine the effect of benzophenone on the growth of Channel Catfish Ovary (CCO) cells, an improved MTT assay is applied in this study. From the research of this paper, we reach the conclusion as follows: Based on the tests through Channel Catfish Ovary (CCO) cell, Ctenpharyngoden Idellus Kindy (CIK) cell and Epithelioma Papulosum Cyprinid (EPC) cell, 14% SDS-DMF(pH=4.7) was choosed as the formazam solvent, and the optimum conditions of this improved MTT assay is: the wavelength is 570nm, the incubation time is 4h, the MTT concentration is 1mg/ml and the inoculation cell number depends on the character of the cell, it increases with the growth speed of the cell. The cytotoxicity of benzophenone and estrogenic activity of its metabolites have been studied in cultured Channel Catfish Ovary (CCO) cell line respectively. The live cell numbers were measured by the improved MTT assay. The study shows a restrain effect of benzophenone on CCO when it mixed with culture medium (the living cell number reduces to 72-91% of the normal condition). But its metabolites by Channel Catfish primary hepatocyte have a promotion effect o