氣-液界面暴露汽車尾氣致BEAS-2B細(xì)胞的毒性效應(yīng)
發(fā)布時(shí)間:2018-01-15 12:29
本文關(guān)鍵詞:氣-液界面暴露汽車尾氣致BEAS-2B細(xì)胞的毒性效應(yīng) 出處:《衛(wèi)生研究》2017年05期 論文類型:期刊論文
更多相關(guān)文章: 汽車尾氣 氣-液界面 人肺細(xì)胞 環(huán)境毒理
【摘要】:目的應(yīng)用新型的氣-液界面細(xì)胞染毒手段,探討汽車尾氣對(duì)人肺細(xì)胞毒性效應(yīng)的影響程度,以及采用0.2μm濾器過(guò)濾尾氣干預(yù)后的細(xì)胞毒性變化情況。方法用20 L Tedlar氣袋收集汽車尾氣,并通過(guò)粒徑譜儀分析0.2μm濾器過(guò)濾前后的汽車尾氣顆粒物數(shù)量、表面積和質(zhì)量濃度。實(shí)驗(yàn)分為空白對(duì)照組、潔凈空氣組、汽車尾氣過(guò)濾組和汽車尾氣組。空白對(duì)照組未作任何處理;潔凈空氣組為21%O_2和79%N_2;汽車尾氣過(guò)濾組為用0.2μm顆粒物過(guò)濾器過(guò)濾后的汽車尾氣;汽車尾氣組為用氣袋直接收集的汽車尾氣。除了空白對(duì)照組,其余3組利用氣-液界面體外染毒技術(shù),在染毒流量為25 mL/min,37℃水浴條件下,對(duì)生長(zhǎng)在氣-液界面插件多孔膜上的BEAS-2B細(xì)胞持續(xù)染毒60 min。利用CCK-8(cell counting Kit-8)檢測(cè)法評(píng)價(jià)細(xì)胞的存活率。采用2',7'-二氯熒光黃雙乙酸鹽(2',7'-dichloro-dihydro-fluorescein diacetate,DCFH-DA)探針檢測(cè)細(xì)胞內(nèi)活性氧(reactive oxygen species,ROS)水平。Annexin V-FITC/PI細(xì)胞凋亡檢測(cè)試劑盒檢測(cè)汽車尾氣對(duì)BEAS-2B細(xì)胞凋亡及壞死率的影響。結(jié)果汽車尾氣組中0.5~10μm顆粒物數(shù)量濃度、表面積濃度和質(zhì)量濃度分別為0.24×10~3個(gè)/cm~3、0.29×10~3μm~2/cm~3和0.19μg/m~3,10~500 nm顆粒物數(shù)量濃度、表面積濃度和質(zhì)量濃度分別為56×10~3個(gè)/cm~3、34.53×10~8nm~2/cm~3和95 ng/m~3。汽車尾氣過(guò)濾組0.5~10μm顆粒物數(shù)量濃度、表面積濃度和質(zhì)量濃度分別低于1個(gè)/cm~3、1μm~2/cm~3和0.001μg/m~3,10~500 nm顆粒物數(shù)量濃度、表面積濃度和質(zhì)量濃度與汽車尾氣組相比分別降低了89.79%、93.57%和90.55%。潔凈空氣組的細(xì)胞存活率、胞內(nèi)ROS、早期凋亡率和晚期凋亡壞死率分別為(90.15±4.25)%、(1.92±0.34)×10~5、(1.09±0.48)%、(8.93±3.31)%。與潔凈空氣組比較,兩組汽車尾氣組的細(xì)胞存活率、胞內(nèi)ROS和晚期凋亡及壞死率差異均有統(tǒng)計(jì)學(xué)意義(P0.05)。汽車尾氣過(guò)濾組與汽車尾氣組之間的比較發(fā)現(xiàn),汽車尾氣組細(xì)胞存活率[(58.09±4.06)%]低于汽車尾氣過(guò)濾組[(66.85±3.49)%],差異有統(tǒng)計(jì)學(xué)意義(t=6.331,P0.001),胞內(nèi)ROS[汽車尾氣過(guò)濾組∶汽車尾氣組=(2.94±0.21)×10~5∶(3.32±0.49)×10~5,t=-1.252,P=0.279]、早期凋亡率[汽車尾氣過(guò)濾組∶汽車尾氣組=(1.09±0.30)%∶(0.99±0.10)%,t=0.708,P=0.497]和晚期凋亡壞死率[汽車尾氣過(guò)濾組∶汽車尾氣組=(21.75±10.37)%∶(15.32±2.74)%,t=1.347,P=0.242]差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論人肺細(xì)胞氣-液界面暴露于汽車尾氣可產(chǎn)生顯著的毒性效應(yīng),0.2μm過(guò)濾器過(guò)濾尾氣干預(yù)后,細(xì)胞毒性會(huì)明顯降低。
[Abstract]:Objective to investigate the effect of automobile exhaust on the cytotoxicity of human lung cells by means of a new method of cell exposure at the gas-liquid interface. And the changes of cytotoxicity after the treatment with 0.2 渭 m filter filter. Methods the vehicle exhaust was collected by 20L Tedlar air bag. The quantity, surface area and mass concentration of vehicle exhaust particles before and after filtration with 0.2 渭 m filter were analyzed by particle size spectrometer. The experiment was divided into blank control group and clean air group. Vehicle exhaust filtration group and vehicle exhaust group. Blank control group did not do any treatment; In the clean air group, there were 21 O / s _ 2 and 79 / N _ 2; The vehicle exhaust was filtered by 0.2 渭 m particulate matter filter. The vehicle exhaust group was collected directly by air bag. In addition to the blank control group, the other three groups were exposed to the gas liquid interface in vitro under the water bath condition of 25 mL 路min ~ (-1) 路min ~ (-1) ~ 37 鈩,
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