大气PM_(2.5)对人支气管上皮细胞增殖和凋亡的影响

Effect of atmospheric PM_(2.5) on cell proliferation and apoptosis in human bronchial epithelial cells

目的探讨PM2.5暴露后,人支气管上皮细胞(16HBE)的增殖和凋亡的变化,研究PM2.5对16HBE细胞的影响。方法将16HBE细胞暴露于0、50、100、200、400、600、800、1 000μg/mL PM2.5悬浮液,24 h后加入CCK-8,3～4 h后用酶标仪在450 nm处测量OD值,并计算细胞存活率。将16HBE细胞暴露于0、50、100、200、400、800μg/mL PM2.5悬浮液,24 h后加入hoechst33342染色25 min、PI染色30 min,显微镜下观察调亡的细胞并计数,计算细胞凋亡率。结果 PM2.5暴露24 h后,50、100、200、400、600、800、1 000μg/mL组细胞存活率显著下降,分别为(89.41±5.53)%、(75.71±5.99)%、(82.99±15.09)%、(61.74±9.88)%、(57.23±3.28)%、(46.00±4.23)%、(42.63%±7.79)%,有剂量效应关系,与对照组相比,差异均具有统计学意义(P均<0.01)。PM2.5对16HBE细胞的24 h半数致死浓度(LC50)为(762.03±180.25)μg/mL。PM2.5暴露24 h后,在镜下计数各剂量组调亡细胞,细胞凋亡数量随着剂量升高而增高,0、50、100、200、400、800μg/mL组细胞调亡率分别为(10.50±2.64)%、(17.90±4.63)%、(21.30±2.26)%、(32.10±4.18)%、(36.90±3.25)%、(50.30±5.81)%,有剂量效应关系,与对照组相比,差异均具有统计学意义(P均<0.01)。结论 PM2.5对16HBE细胞有毒性作用,可导致16HBE细胞增殖受到抑制,凋亡增加。

Objective To investigate changes in cell proliferation and apoptosis of 16 HBE cells after PM2.5exposure, andto earch effects of PM2.5on 16 HBE cells. Methods 16 HBE cells were exposed to PM2.5in the concentration of 0, 50, 100, 200,400, 600, 800, 1 000 μg/mL, 24 h after exposure, CCK-8 was added and OD value was detected at 450 nm by enzyme-labeledinstrument, then cell viability was calculated. 16 HBE cells were exposed to PM2.5in the concentration of 0, 50, 100, 200, 400,800 μg/mL, 24 h after exposure, cells were stained with hoechst33342 for 25 min and PI for 30 min. Cell apoptosis wasobserved and counted under microscope, then the cell apoptosis rate was calculated. Results With 24 h after PM2.5exposure,compared with control group, cell viability was significantly decreased,(89.41±5.53)%,(75.71±5.99)%,(82.99±15.09)%,(61.74±9.88)%,(57.23±3.28)%,(46.00±4.23)%,(42.63%±7.79)% in the concentration of 0,50, 100, 200, 400, 600, 800, 1000 μg/mL, respectively(P all<0.01).There was a dose-effect relationship between PM2.5concentration and cell viability.Median lethal concentration(LC50) in 24 h of PM2.5to 16 HBE cells was(762.03 ± 180.25)μg/mL. With 24 h after PM2.5exposure, compared with control group,cell apoptosis was increased significantly with the increase of PM2.5concentration, cellapoptosis rate was(10.50±2.64)%,(17.90±4.63)%,(21.30±2.26)%,(32.10±4.18)%,(36.90±3.25)%,(50.30±5.81)% inthe concentration of 0,50, 100, 200, 400, 800 μg/mL, respectively(P all<0.01). There was a dose-effect relationship betweenPM2.5concentration and cell apoptosis rate.Conclusion PM2.5plays a toxical effect on 16 HBE cells, could induce inhibition ofcells proliferation and caused apoptosis.