南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (01): 17-.

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PRKCD和ASK1在人髓系白血病U937细胞分化过程中的作用

闫峰,王效民,袁思波,马全明,韩慧平   

  • 出版日期:2015-01-20 发布日期:2015-01-20

Role of PRKCD and ASK1 in U937 cell differentiation

  • Online:2015-01-20 Published:2015-01-20

摘要: 目的通过检测ASK1 和PRKCD在单核细胞分化过程中的表达变化,探索其在门静脉高压症(PH)脾亢脾脏巨噬细胞
(MΦ)功能变化中所起的作用。方法采用佛波酯诱导U937 分化成为巨噬细胞样细胞,q-PCR 检测不同分化阶段ASK1 和
PRKCD mRNA的表达,Western Blot 检测诱导前后ASK1和PRKCD蛋白表达水平变化,ELISA法检测细胞分化过程中与吞噬
功能相关的细胞因子IL-10和TNF-α的分泌情况,鸡红细胞吞噬试验鉴定诱导后细胞功能。结果随着PMA诱导U937细胞时
间延长,ASK1和PRKCD基因表达水平下降,TNF-α的分泌量逐渐增加,而IL-10在诱导后24 h达到最大分泌量,随后又呈下降
趋势;Western Blot 结果显示,ASK1及p-ASK1蛋白表达水平较诱导前均显著上升,PRKCD及p-PRKCD蛋白表达水平显著下
降;吞噬实验结果显示,诱导后的细胞具有一定的吞噬功能。结论在PMA诱导的U937细胞向单核细胞分化过程中,ASK1蛋
白表达上调,PRKCD蛋白表达下调,与脾亢脾Mφ一致,并导致Mφ活性增强。

Abstract: Objective To investigate the expression of ASK1 and PRKCD in the process of monocyte differentiation, and explore
their role in functional changes of hypersplenism spleen macrophages (MΦ) in portal hypertension (PH). Methods U937 cells
were stimulated to differentiate into monocyte/macrophage-like cells by cultivation in PMA and the mRNA expressions of
ASK1 and PRKCD were detected by q-PCR and the changes of protein expression were identified by western blot analysis. The
secretion of phagocytose related cytokines such as IL-10 and TNF-a were tested by ELISA, and the function of the
macrophage-like cells were studied by chicken red blood cell phagocytose test. Results The expressions of PRKCD and ASK1
mRNA were gradually decreased along with the cell differentiation, while the secretion of TNF-α was increased, IL-10
secretion reached a maximum at 24 h after PAM stimulation, and then gradually fell. The expression of ASK1 and p-ASK1
were rapidly increased compared with the non-stimulated U937 cells, while the expression of PRKCD and p-PRKCD were
sightly declined. The phagocytose test show that U937 cells induced with PMAwere able to swallow the chicken red blood cell.
Conclusion Up-regulated protein expression of ASK1 and p-ASK1 and down-regulated protein expression of PRKCD and
p-PRKCD in the process of PMA induced monocyte differentiation, are consist with the expression changes of splenic
macrophage phagocytosis in hypersplenism, which leads to increased activity of MΦ.