槲皮素对PM 2.5导致支气管上皮细胞炎症的保护作用

Protective effect of quercetin on inflammation of bronchial epithelial cells induced by PM2.5

目的:考察槲皮素对PM2.5刺激支气管上皮细胞(BEAS-2B)后细胞炎症的保护作用。方法:通过MTT法检测不同浓度槲皮素作用下细胞的存活率;通过Griess法检测槲皮素对PM2.5刺激后BEAS-2B细胞一氧化氮(NO)释放量;通过酶联免疫吸附法检测炎症因子TNF-α,IL-1β,IL-6含量;通过蛋白免疫印迹法检测NF-κB p65的表达量。结果:细胞实验表明:与对照组相比,200μg/ml及以上浓度的PM2.5能显著降低细胞的存活率,在PM 2.5的作用下,细胞内NO和炎症因子的释放量均显著增加,同时转录因子NF-κB p65和受体TLR4的表达量也显著增加,而抑制蛋白IκB-α的含量降低。经槲皮素处理后,30,100μM槲皮素能够显著恢复PM2.5所致的细胞存活率降低;100μM槲皮素能降低NO的释放量;30、100μM槲皮素降低炎症因子IL-1β和IL-6的释放,以及TLR4的蛋白表达量;3、30、100μM槲皮素降低炎症因子TNF-α释放和NF-κB p65的蛋白表达,同时升高IκB-α的含量。结论:槲皮素对PM2.5导致的支气管上皮细胞炎症具有显著的保护作用,其作用机制可能与NF-κB通路相关。

Objective： To investigate the protective effects of quercetin on bronchial epithelial cells（ BEAS-2B） inflammation stimulated by PM 2.5.Methods： The survival rates of cells treated with different concentrations of quercetin were detected by MTT method.The release of NO in BEAS-2B cells after PM 2.5stimulation was measured by Griess method.The levels of inflammatory factors,such as TNF-α,IL-1β and IL-6,were detected by enzyme-linked immunosorbent assay.The protein expression of NF-κB p65 was detected by Western blotting.Results：Experimental results showed that greater than and equal to 200 μg/ml PM 2.5 significantly decreased the cell survival rate in contrast to the control group（ P 〈 0.05）.The concentration of NO and inflammatory factors（ P 〈 0.01）,as well as the expression of NF-κB and TLR4（ P 〈0.05）,increased significantly after PM2.5 stimulation,while the content of inhibition protein IκB-α decreased（ P 〈 0.01）.After the treatment with different quercetin,30,100 μM quercetin could restore the decrease of cell viability（ P 〈 0.01）; 100 μM quercetin could reduce the NO release（ P 〈 0.01）; 30,100 μM quercetin could reduce the release of inflammatory factors IL-1β and IL-6,as well as the expression of TLR4（ P 〈 0.05）; 3,30,100 μM quercetin could reduce the release of TNF-α inflammatory factors（ P 〈 0.01） and NF-κB（ P 〈 0.05）,at the same time increase IκB-α（ P 〈 0.05）.Conclusion： Quercetin has protective effect on PM2.5-induced inflammation of bronchial epithelial cells,and its mechanism may be related to NF-κB pathway.