【目的】研究瞬时过表达锌指基因ZNF580(zinc finger factor-8)对白介素-8(interleukin-8)的表达以及单核细胞趋化功能的影响。为探讨ZNF580在炎症反应中的机制提供依据。【方法】重组质粒p EGFP-ZNF580经转染试剂Lipofectamine TM2000瞬时转染内皮细胞融合株EA.hy926,通过鉴定高表达ZNF580后采用Real-time PCR检测IL-8的m RNA表达水平以及酶联免疫吸附法检测过表达ZNF580细胞上清中IL-8的分泌情况,运用transwell小室趋化实验分析高表达ZNF580对单核细胞THP-1细胞趋化功能的影响差别。【结果】瞬时过表达ZNF580的内皮细胞构建成功,高表达ZNF580后内皮细胞IL-8 m RNA表达及上清中分泌水平均上调,同时趋化实验证实,过表达ZNF580可引起THP-1细胞趋化功能的明显差异。【结论】构建瞬时过表达ZNF580对内皮细胞IL-8的表达及THP-1细胞趋化功能有重要促进作用,初步揭示ZNF580在动脉粥样硬化炎症的发生中的重要作用。
【Objective】To study the effect of overexpression of ZNF580 in expression of interleukin-8 and chemotaxis in monocyte.【Methods】Recombinant plasmid p EGFP-ZNF580 transient transfected by Lipofectamine TM2000 to endothelial cell line EA.hy926,after identification high expression of ZNF580, the m RNA expression of IL-8 was detected by real-time PCR, the secreted IL-8 was determined by ELISA, The chemotaxis in monocyte cells was evaluated by transwell migration.【Results】Transient high expression of ZNF580 were succeed to construction. Transient transfection of ZNF580 resulted in an increase of the cytokine IL-8 in m RNA and protein level, which also showed significantly discrepancy in motility of human acute monocytic leukemia cells(THP-1) by transwell bioassay.【Conclusion】Overexpression of ZNF580 plays a critical role in augmenting the release of IL-8 and chemotactic activity of THP-1cells. which preliminary demonstrate that it may be a new potential target for intervention in atherosclerosis inflammation.