4种方法检测大气颗粒物PM2.5的细胞毒性
发布时间：2019-08-14 20:12
【摘要】：目的采用多种检测原理和观察终点的细胞毒性试验综合评估提取PM2.5的细胞毒性效应,为深入研究PM2.5生物学效应提供实验依据。方法用去离子水提取PM2.5,溶于二甲亚砜获得PM2.5悬液。不同浓度PM2.5染毒支气管上皮(HBE)细胞24 h,采用噻唑蓝比色(MTT)、乳酸脱氢酶释放法(LDH法)、台盼蓝拒染法、吖啶橙-溴化乙锭双染法(AO-EB双染法)4种细胞毒性实验评估PM2.5的细胞毒性,并计算半数抑制浓度(IC_(50))。结果去离子水提取的PM2.5溶于二甲亚砜后呈黑褐色,肉眼可见颗粒成分。不同浓度的PM2.5处理HBE细胞24 h后,台盼蓝拒染法(IC_(50)=815.5μg/m L)、LDH释放法(IC_(50)=827.3μg/m L)及AO-EB双染法(IC_(50)=887.5μg/m L)检测结果与空白对照组和溶剂对照组相比,细胞存活率均显著下降(P0.05);而MTT法的细胞存活率检测结果显著高于对照组1.5~4.0倍(P0.05),与细胞实际生长状态不符。结论用去离子水提取PM2.5可减少溶剂带入,能够最大程度反映空气污染实际情况;实验采用的4种方法都可检测出水提PM2.5细胞毒性,但由于样品颜色的特殊性,MTT实验不能准确检测本研究中PM2.5的细胞毒性。
[Abstract]:Objective to evaluate the cytotoxicity of PM2.5 extraction by using various detection principles and cytotoxicity tests at the end of observation, so as to provide experimental basis for further study of the biological effects of PM2.5. Methods PM2.5, was extracted from deionized water and dissolved in dimethyl sulfoxide to obtain PM2.5 suspension. Bronchial epithelial (HBE) cells were exposed to different concentrations of PM2.5 for 24 h. The cytotoxicity of PM2.5 was evaluated by thiazolyl blue colorimetric (MTT), lactic dehydrogenase release assay (LDH method), trypan blue exclusion assay and AO-EB double staining (AO-EB double staining method), and the half inhibitory concentration (IC_ (50).) was calculated. Results the PM2.5 extracted from deionized water was black brown after dissolving in dimethyl sulfoxide, and the granule composition could be seen by naked eye. After HBE cells were treated with different concentrations of PM2.5 for 24 h, the survival rate of HBE cells treated with trypan blue exclusion assay (IC_ (50) = 815.5 渭 g / m L), LDH) (IC_ (50) = 827.3 渭 g / m L) and AO-EB double staining (IC_ (50) = 887.5 渭 g / m L) was significantly lower than that of the blank control group and solvent control group (P 0.05). The cell survival rate of MTT assay was significantly higher than that of the control group 1.5 鈮，
本文编号：2526771
[Abstract]:Objective to evaluate the cytotoxicity of PM2.5 extraction by using various detection principles and cytotoxicity tests at the end of observation, so as to provide experimental basis for further study of the biological effects of PM2.5. Methods PM2.5, was extracted from deionized water and dissolved in dimethyl sulfoxide to obtain PM2.5 suspension. Bronchial epithelial (HBE) cells were exposed to different concentrations of PM2.5 for 24 h. The cytotoxicity of PM2.5 was evaluated by thiazolyl blue colorimetric (MTT), lactic dehydrogenase release assay (LDH method), trypan blue exclusion assay and AO-EB double staining (AO-EB double staining method), and the half inhibitory concentration (IC_ (50).) was calculated. Results the PM2.5 extracted from deionized water was black brown after dissolving in dimethyl sulfoxide, and the granule composition could be seen by naked eye. After HBE cells were treated with different concentrations of PM2.5 for 24 h, the survival rate of HBE cells treated with trypan blue exclusion assay (IC_ (50) = 815.5 渭 g / m L), LDH) (IC_ (50) = 827.3 渭 g / m L) and AO-EB double staining (IC_ (50) = 887.5 渭 g / m L) was significantly lower than that of the blank control group and solvent control group (P 0.05). The cell survival rate of MTT assay was significantly higher than that of the control group 1.5 鈮，
本文编号：2526771
本文链接：https://www.wllwen.com/yixuelunwen/yufangyixuelunwen/2526771.html
