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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 effect of quercetin on cellular inflammation after PM2.5 stimulation of bronchial epithelial cells (BEAS-2B). Methods: MTT assay was used to test the survival rate of cells under different concentrations of quercetin. The release of nitric oxide (NO) from BEAS-2B cells after quercetin treatment with quercetin was detected by Griess method. The levels of NO were detected by enzyme-linked immunosorbent assay The levels of TNF-α, IL-1β and IL-6 were detected by Western blotting. The expression of NF-κB p65 was detected by Western blotting. Results: The results of cell experiments showed that compared with the control group, the PM2.5 concentration of 200μg / ml and above significantly reduced the cell viability. The release of NO and inflammatory cytokines increased significantly under PM 2.5, At the same time, the expression of transcription factors NF-κB p65 and TLR4 also increased significantly, but the content of IκB-α decreased. After treated with quercetin, quercetin at 30,100μM could significantly reduce the cell viability decreased by PM2.5; 100μM quercetin can reduce the release of NO; 30,100μM quercetin can reduce inflammatory cytokines IL-1β and IL-6 release and TLR4 protein expression; 3,30,100μM quercetin reduce the release of inflammatory cytokines TNF-α and NF-κB p65 protein expression, while increasing IκB-α content. Conclusion: Quercetin has a significant protective effect on the inflammation of bronchial epithelial cells induced by PM2.5, and its mechanism may be related to NF-κB pathway.