大气细颗粒物(PM_(2.5))对肥大细胞分泌β-氨基己糖苷酶、组胺和IL-4的影响

Effect of atmospheric fine particulate matter( PM_(2.5)) on secretion of β-hexosaminidase,histamine and interleukin-4 in mast cells in vitro

目的探究大气细颗粒物(PM_(2.5))对肥大细胞的生物学效应。方法采用智能中流量空气总悬浮微粒采样器采集重庆市内非工业区空气中PM_(2.5),以不同浓度(0、12.5、25.0、50.0、100.0、200.0、400.0、800.0μg/m L)PM_(2.5)混悬液作用于人肥大细胞系LAD2,采用微板法检测LAD2细胞β-氨基己糖苷酶的释放水平;采用ELISA法检测LAD2细胞释放组胺、白细胞介素4(interleukiin-4,IL-4)以及活性氧(reactive oxygen species,ROS)水平。结果终浓度为25.0μg/m L的PM_(2.5)刺激后,LAD2细胞分泌并释放入细胞上清的β-氨基己糖苷酶、组胺及IL-4的水平高于0μg/m L PM_(2.5)组[(23.78±3.98)%vs(14.05±1.12)%,(12.32±0.18)vs(9.72±0.23)μg/m L,(267.63±7.97)vs(154.25±7.71)pg/m L,P<0.01];12.5~100.0μg/m L PM_(2.5)刺激后,LAD2细胞产生ROS水平逐渐升高,终浓度为25.0μg/m L的PM_(2.5)刺激LAD2细胞后产生的ROS水平高于0μg/m L PM_(2.5)组[(7.99±0.29)vs(5.88±0.49)ng/m L,P<0.01]。结论 PM_(2.5)可能通过氧化应激致使肥大细胞产生ROS,引起肥大细胞产生细胞因子和炎症因子。

Objective To investigate the biological effect of atmospheric fine particulate matter （PM2.5） on mast cells in vitro. Methods PM25 was collected in the nonindustrial zone of Chongqing city using an intelligent medium-volume total suspended particulate air sampler and prepared into mixed suspensions of different concentrations （0, 12.5, 25.0, 50.0, 100.0, 200.0, 400.0 and 800.0 Ixg/mL）. Human mast cell line LAD2 exposed to these suspensions were examined for cellular production of 13-hexosaminidase using microplate method and for the release levels of histamine, interleukin 4 （ IL-4 ） and reactive oxygen species （ROS） using ELISA. Results Exposure of the mast cells to 25 μg/mL PM2.5 resulted in significant increases in the levels of β-hexosaminidase [ （ 23.78 ± 3.98 ） % vs （ 14.05 ± 1.12 ） % , P〈0.01], histamine （12.32±0. 18 vs9.72±0.23 μg/mL, P〈0.01） and IL-4 （267.63 ±7.97 vs 154. 25±7.71 pg/mL, P 〈 0. 01 ） in the supernatant as compared with the cells without PM2.5 exposure. Within the concentration range from 12.5 to 100.0 g/mL, an increasing concentration of PM2.5 suspension caused progressive increase in ROS levels in the cells, and the cells exposed to 25 g/mL of the suspension showed a significantly higher ROS level than the cells without PM2.5 exposure （7.99 ±0.29 vs 5.88 ±0. 49 ng/mL, P 〈0. 01）. Gonclusion PM2.5 exposure may induce ROS production via oxidative stress to promote the production of cytokines and inflammatory factors in LAD2 cells in vitro.