纳米氧化锌颗粒物诱导人支气管上皮细胞白细胞介素-8基因的表达及调控机制

Inductive eft ect of zinc oxide nanoparticles on interleukin 8 gene expression in human bronchial epithelial cells and its regulatory mechanism

目的阐明纳米氧化锌颗粒ZnO-NPs（30rim）对人支气管上皮细胞（BEAS-2B）白细胞介素-8（IL-8）基因表达的影响及调控机制。方法利用BEAS-2B细胞作为研究对象，采用噻唑蓝（Mrr）法测定ZnO-NPs对细胞的损伤作用。应用RT-PCR技术及ELISA法检测纳米氧化锌颗粒物对细胞内IL-8mRNA和蛋白表达的影响。利用IL-8mRNA降解试验测定ZnO-NPs对转录后IL-8mRNA稳定性的影响。结果ZnO-NPs刺激可致细胞内IL-8mRNA水平及培养液上清中IL-8蛋白含量明显升高。转录抑制剂可显著降低ZnO-NPs对IL-8mRNA表达的诱导作用。在用放线菌素D终止IL-8mRNA合成的前提下，ZnO-NPs可明显减缓细胞内IL-8mRNA的降解。结论纳米氧化锌颗粒物可提高IL-8mRNA及蛋白在BEAS-2B内的表达水平；ZnO-NPs对细胞内IL-8mRNA具有稳定作用。

Objective To clarify the effect of zinc oxide nanoparticles （ZnO-NPs） （30 nm in diameter） on the interleukin 8 （IL-8） gene expression in human bronchial epithelial cells （BEAS-2B） and its regulatory mechanism. Methods BEAS-2B cells were used in the study. The MTY assay was employed to evaluate the damage to BEAS-2B cells by ZnO-NPs. RT-PCR and ELISA were used to measure the mRNA and protein expression levels of IL-8 in the BEAS-2B ceils exposed to ZnO-NPs. The IL-8 mRNA decay assay was used to determine the effect of ZnO-NPs on IL-8 mRNA stability. Results Exposure to ZnO-NPs significantly increased the level of IL-8 mRNA in BEAS-2B cells and the level of IL-8 protein in supernatant medium. The transcription inhibitor significantly reduced the mRNA expression of IL-8 induced by ZnO-NPs. ZnO-NPs significantly delayed IL-8 mRNA degradation in the BEAS-2B cells that were pretreated with actinomycin D for terminating IL-8 mRNA synthesis. Conclusion ZnO-NPs can increase the mRNA and protein expression levels of IL-8 and IL-8 mRNA stability in BEAS-2B cells.