4種方法檢測大氣顆粒物PM2.5的細(xì)胞毒性
發(fā)布時(shí)間：2019-08-14 20:12
【摘要】：目的采用多種檢測原理和觀察終點(diǎn)的細(xì)胞毒性試驗(yàn)綜合評估提取PM2.5的細(xì)胞毒性效應(yīng),為深入研究PM2.5生物學(xué)效應(yīng)提供實(shí)驗(yàn)依據(jù)。方法用去離子水提取PM2.5,溶于二甲亞砜獲得PM2.5懸液。不同濃度PM2.5染毒支氣管上皮(HBE)細(xì)胞24 h,采用噻唑藍(lán)比色(MTT)、乳酸脫氫酶釋放法(LDH法)、臺盼藍(lán)拒染法、吖啶橙-溴化乙錠雙染法(AO-EB雙染法)4種細(xì)胞毒性實(shí)驗(yàn)評估PM2.5的細(xì)胞毒性,并計(jì)算半數(shù)抑制濃度(IC_(50))。結(jié)果去離子水提取的PM2.5溶于二甲亞砜后呈黑褐色,肉眼可見顆粒成分。不同濃度的PM2.5處理HBE細(xì)胞24 h后,臺盼藍(lán)拒染法(IC_(50)=815.5μg/m L)、LDH釋放法(IC_(50)=827.3μg/m L)及AO-EB雙染法(IC_(50)=887.5μg/m L)檢測結(jié)果與空白對照組和溶劑對照組相比,細(xì)胞存活率均顯著下降(P0.05);而MTT法的細(xì)胞存活率檢測結(jié)果顯著高于對照組1.5~4.0倍(P0.05),與細(xì)胞實(shí)際生長狀態(tài)不符。結(jié)論用去離子水提取PM2.5可減少溶劑帶入,能夠最大程度反映空氣污染實(shí)際情況;實(shí)驗(yàn)采用的4種方法都可檢測出水提PM2.5細(xì)胞毒性,但由于樣品顏色的特殊性,MTT實(shí)驗(yàn)不能準(zhǔn)確檢測本研究中PM2.5的細(xì)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，
本文編號：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，
本文編號：2526771
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