阿特拉津对小鼠腹腔巨噬细胞功能影响的体外研究

Effects of atrazine on function of murine peritoneal macrophages in vitro

目的探讨阿特拉津（ATZ）体外染毒对小鼠腹腔巨噬细胞功能的影响,为其免疫毒性评价积累资料。方法分离培养小鼠腹腔巨噬细胞,经不同浓度（0~1000μmol/L）阿特拉津处理后,MTT比色法测定细胞活力,中性红（NR）法测定其吞噬功能,硝酸还原酶法测定NO含量,ELISA法测定TNF-α释放能力,DCFH-DA荧光标记流式细胞术测定活性氧含量。结果 MTT法测得ATZ对小鼠腹腔巨噬细胞的24、48、72 h染毒的IC50分别为（887.4±1.11）、（360.2±1.13）、（270.6±1.20）μmol/L。NR吸收法所测得各时间点IC50分别为（214.2±1.14）、（120.0±1.10）、（42.60±1.12）μmol/L。24 h及72 h染毒后,低浓度ATZ剂量组NO含量较对照组增加,高浓度剂量组NO含量较对照组出现下降趋势。72 h染毒后,低浓度ATZ剂量组（0.01、0.1μmol/L）TNF-α的含量高于对照组（P〈0.05）,但在1μmol/L及以上的ATZ染毒组TNF-α含量较对照组降低（P〈0.05）。小鼠腹腔巨噬细胞在ATZ低剂量组（0.01、1μmol/L）染毒24 h和72 h时均能促使ROS的产生（P〈0.05）,但在100μmol/L剂量组里ROS释放量却降低。结论阿特拉津染毒可影响小鼠腹腔巨噬细胞的活性和吞噬功能,并可干扰其细胞因子/活性氧/活性氮的释放。

Objective To investigate the effects of atrazine（ ATZ） on function of peritoneal macrophages in vitro,and to provide data for its immunotoxicity evaluation.Methods The murine peritoneal macrophages were isolated from mice. After treated with ATZ at different doses,cells viability was detected by MTT assay,and phagocytosis was evaluated by neutral red absorption. NO and TNF-α release were detected with nitrate reductase method and ELISA method,respectively. ROS was measured by DCFH-DA and flow cytometry. Results The IC50 values of the peritoneal macrophages for the MTT assay were（ 887. 4 ± 1. 11）,（ 360. 2 ± 1. 13） and（ 270. 6 ± 1. 20） μmol / L after 24,48 and72 h. While for the NR absorption were（ 214. 2 ± 1. 14）,（ 120. 0 ± 1. 10） and（ 42. 60 ±1. 12） μmol / L. After dosed for 24 h and 72 h,the NO release of low dose groups were higher than those of the controls,while the NO release of high dose groups showed adownward trend when compared with the control＇s. After treatment for 72 h,the TNF-αsecretion of low dose groups（ 0. 01,0. 1 μmol / L） were significantly higher than that of the control（ P〈0. 05）,and TNF-α secretion in the other groups were lower when compared with that of the control（ P〈0. 05）. After exposure for 24 h or 72 h,the ROS release was promoted in both 0. 01 and 1 μmol / L ATZ groups（ P〈0. 05）,however,the ROS release were lowered in the 100 μmol / L ATZ group. Conclusion ATZ can affect the cells viability and phagocytosis of peritoneal macrophages. Meanwhile the TNF-α secretion,NO release as well as ROS release are disturbed in peritoneal macrophages exposed to ATZ.