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目的探讨并比较不同粒径汽车尾气颗粒物对人肺癌上皮细胞A549肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)、核转录因子-κB(NF-κB)的mRNA及NF-κB蛋白表达的影响。方法以不同粒径(0.18~3.2μm)汽车尾气颗粒物混悬液(100μg/ml)染毒A549细胞,48 h后RT-PCR法检测细胞TNF-α、IL-6及NF-κB(P65)基因表达水平,Western Blot法检测胞浆及胞核内NF-κB(P65)蛋白表达水平。结果与对照组相比,各染毒组细胞的TNF-α、IL-6及NF-κB(P65)mRNA表达水平均上升(P<0.05),由高到低依次为0.56~、0.32~、0.18~、1.0~和1.8~3.2μm组,两两之间(除0.18~和0.32~μm组之间、1.0~和1.8~3.2μm组之间)差异均有统计学意义(P<0.05);各组TNF-α、IL-6 mRNA水平与NF-κB(P65)mRNA水平显著相关(r=0.944,0.890,P<0.05)。与对照组相比,除1.8～3.2μm组无统计学意义外,其余各组胞浆NF-κB(P65)蛋白水平均降低(P<0.05),由高到低依次为1.0~、0.18~、0.32~、0.56~μm组;而胞核NF-κB(P65)蛋白水平均升高(P<0.05),由低到高依次为1.0~、0.18~、0.32~、0.56~μm组。结论不同粒径汽车尾气颗粒物作用于A549细胞后均可使TNF-α、IL-6、NF-κB mRNA表达升高、NF-κB活化,活化的NF-κB由细胞质进入细胞核发挥其调控炎症反应的作用。各染毒组中以0.56~μm组炎性损伤效应最强,粒径是影响颗粒物毒性作用大小的重要因素之一。
Objective To investigate and compare the effect of vehicle particulate matter of different particle sizes on the mRNA and protein expression of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and nuclear transcription factor-κB -κB protein expression. Methods A549 cells were exposed to vehicle tail suspension (100μg / ml) with different particle sizes (0.18 ~ 3.2μm). The expression of TNF-α, IL-6 and NF- The expression of NF-κB (P65) protein in cytoplasm and nucleus was detected by Western Blot. Results Compared with the control group, the expression levels of TNF-α, IL-6 and P65 mRNA in all the treated groups increased (P <0.05), and the highest and lowest were 0.56 ~ 0.32 ~ 0.18 ~, 1.0 ~ and 1.8 ~ 3.2μm group, the difference was statistically significant (P <0.05) between the two groups except the 0.18 ~ and 0.32 ~ μm groups, 1.0 ~ ; The level of TNF-α and IL-6 mRNA in each group was significantly correlated with the level of NF-κB (P65) mRNA (r = 0.944,0.890, P <0.05). Compared with the control group, the protein levels of NF-κB (P65) in cytoplasm decreased in all groups except the 1.8 ~ 3.2μm group (P <0.05), the descending order was 1.0 ~ 0.18 ~ , 0.32 ~, 0.56 ~ μm group, while the protein level of nuclear NF-κB (P65) increased (P <0.05) from 1.0 to 0.18 ~ 0.32 ~, 0.56 ~ μm. CONCLUSION: The effects of vehicle particulate matter of different particle size on A549 cells can increase the expression of TNF-α, IL-6 and NF-κB mRNA and activate NF-κB. The activated NF-κB enters the nucleus from the cytoplasm to regulate its inflammatory response Role. The 0.56 ~ μm group showed the strongest inflammatory injury effect in each group, and the particle size was one of the important factors affecting the toxic effect of particulate matter.