NF-κB 信号通路在PM2.5促进肺上皮细胞黏蛋白Muc5ac分泌中的作用
首发时间:2019-04-08
摘要:目的:探讨NF-κB信号通路在大气PM2.5影响肺上皮细胞黏蛋白Muc5ac分泌的作用。方法:采用含10%胎牛血清的DMEM培养基体外培养人肺上皮细胞(Beas-2B细胞),分别给予空白对照组、低剂量组、中剂量组、高剂量组0、100 、200、400μg/mL PM2.5暴露。大气PM2.5暴露6h、12h、24h,收集细胞及培养基上清液。采用MTT法检测Beas-2B细胞的存活率;ELISA法测定培养基上清中IL-1β、TNF-α、Muc5ac的表达水平;Western Blot法检测IκB-α蛋白表达量。结果:PM2.5暴露12、24h,细胞存活率随暴露剂量升高而下降。暴露6、12h后,高剂量组细胞Muc5ac表达水平明显高于其余各组。暴露6、12、24h后,高剂量组IL-1β水平高于中剂量和对照组。暴露6h后,高剂量组TNF-α水平明显高于其余各组,中剂量组TNF-α水平明显高于对照组;暴露24h后,高、中剂量组TNF-α水平明显高于低剂量组,各剂量组TNF-α水平明显高于对照组。随着PM2.5染毒剂量的增加和染毒时间的延长,Beas-2B细胞IκB-α蛋白表达水平逐渐降低。结论:大气PM2.5暴露可降低Beas-2B细胞存活率,并可通过促进细胞NF-κB信号通路活化,刺激细胞分泌更多的黏蛋白和炎性因子。
关键词: NF-κB信号通路 大气颗粒物 黏蛋白Muc5ac 炎症因子
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The Role of NF-κB in Muc5ac secretion of Beas-2B cell induced by PM2.5
Abstract:Objective To explore the role of NF-κB signal pathway in the Muc5ac secretion of Beas-2B cell induced by atmospheric PM2.5. Methods Human lung epithelial cell (Beas-2B cell) was cultured in vitro using DMEM medium containing 10% fetal bovine serum and exposed to 0, 100, 200, 400 μg/mL atmospheric PM2.5 for 6h, 12h, and 24h.The cell and supernatant were collected. The survival rate of Beas-2B cell was detected by MTT assay. The levels of IL-1β, TNF-α and the expression levels of Muc5ac were determined by ELISA. The expression of IκB-α protein was detected by Western Blot.Results After exposure to PM2.5 for 12 and 24 hours,the cell survival rate was decreased with increasing exposure dose. After exposure to PM2.5 for 6 and 12 hours, the expression levels of Muc5ac in high dose group were significantly higher than the other groups. After exposure to PM2.5 for 6, 12, and 24 hours, the levels of IL-1β in high dose group were higher than the middle dose and control groups. After exposure to PM2.5 for 6 hours , the levels of TNF-α in the high dose group were significantly higher than those in the other groups. The levels of TNF-α in the middle dose group were significantly higher than those in the control group. After exposure to PM2.5 for 24 hours, the levels of TNF-α in the high and middle dose groups were significantly higher than those in the low dose group. The levels of TNF-α in each dose groups were significantly higher than those in the control group. The expression level of IκB-α protein in Beas-2B cells gradually decreased with the increase of PM2.5 dose and prolonged exposure time.The expression levels of IκB-α protein in Beas-2B cells were gradually decreased with the increasing of PM2.5 dose and exposure time. Conclusion Exposure to Atmospheric PM2.5 could reduce the survival rate of Beas-2B cells and stimulate the cells to secrete more mucin and inflammatory factors by promoting the activation of NF-κB signal pathway.
Keywords: NF-κB signal pathway atmospheric particulate matter Muc5ac inflammatory factor
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