摘要
[目的]研究乙酸豆寇佛波酯(PMA)诱导人单核细胞系(THP-1)细胞株向巨噬细胞分化过程中核因子kappa B(nuclear factor-kappa B,NF-κB)表达量的变化。[方法]体外利用PMA诱导THP-1细胞分化为巨噬细胞,利用倒置显微镜观察细胞形态学变化,流式细胞仪检测巨噬细胞表面特异性抗原F4/80、CD14。利用Western Blotting检测单核细胞向巨噬细胞分化过程中NF-κB蛋白表达的变化。[结果]经PMA诱导后,THP-1细胞由类圆形、分散、悬浮转变为不规则、黏附、贴壁,且出现F4/80、CD14的高表达。THP-1单核细胞向巨噬细胞分化过程中NF-κB蛋白表达明显增加。[结论]经体外诱导分化成的巨噬细胞中NF-κB蛋白表达与单核细胞比较显著增加,为进一步研究巨噬细胞活化中NF-κB信号通路与颈椎病的关系提供模型。
[Objective] To observe the protein expression of nuclear factor( NF)-κB during THP-1 differentiation,monocytes were induced by phorbol 12-myristate 13-acetate( PMA) to differentiate into macrophages. [Methods] THP-1cells were differentiated to macrophages in vitro by induction with PMA. An inverted microscope was used to observe morphological changes of the differentiated cells. Specific antigen F4 /80 and CD14 of macrophage cells were measured by flow cytometry( FCM) to verify induction. Protein expression of NF-κB was determined by Western blotting. [Results] After induction with PMA,the morphology of THP-1 cells changed from round,dispersed,and suspended to irregular,adhesive,and adherent with high expression of F4 /80 and CD14. NF-κB protein level increased during THP-1 monocyte on version into macrophages. [Conclusion] Protein expression of NF-κB was significantly increased during THP-1 monocyte induction into macrophages. This model can be used to examine the connection between macrophages and cervical spondylosis.
引文
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