【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(IC₅₀)。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(IC₅₀=815.5μg/m L)、LDH释放法(IC₅₀=827.3μg/m L)及AO-EB双染法(IC₅₀=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（P<0.05）;而MTT法的细胞存活率检测结果显著高于对照组1.5⁴.0倍（P<0.05）,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。更多还原

【Abstract】 Objective Four different cytotoxicity test methods were adpoted to evaluate the cytotoxicity of deionized water extracts of PM2.5 on human bronchial epithelium cell line（HBE cell）. Methods PM2.5 was extracted with deionized water by ultrasound water bath. Dried PM2.5 extracts were resolved in DMSO to make the stock solution of 100mg/m L. HBE cells were treated with 0-1000μg/m L PM2.5 extracts for 24 h. Then MTT assay, LDH assay, Trypan blue assay and AO-EB double stain assay were utilized to test cell viability. Results The results of Trypan blue assay(IC₅₀=815.5μg/m L), LDH assay(IC₅₀=827.3μg/m L) and AO-EB double stain assay(IC₅₀=887.5μg/m L) showed that cell viability significantly decreased with increasing PM2.5 concentration in a dose-dependent manner, while abnormal results were observed in MTT assay which was not consistent with the cellular morphology observation. Conclusion Deionized water extracts of PM2.5 could represent the origin fine particulate matter. The testing results of our cytotoxicity methods indicates that MTT assays is not suitable for evaluating cytotoxicity of PM2.5 extracted in deionized water.更多还原