摘要
目的研究单核细胞向泡沫细胞诱导分化过程中脂酰CoA胆固醇酯酰转移酶-1(ACAT-1)表达及活性的变化。方法复苏、培养人单核细胞株THP-1细胞,与乙酸肉豆蔻佛波酯(PMA)、PMA+氧化低密度脂蛋白(ox-LDL)共孵育1～4 d,RT-PCR方法检测单核细胞分化过程中ACAT-1 mRNA表达,Westernblotting检测ACAT-1蛋白表达的变化。结果(1)单核细胞复苏后加入PMA诱导48 h后,分化为巨噬细胞。经PMA+ox-LDL诱导24 h后分化为泡沫细胞。(2)与无血清培养单核细胞对照组比较,在PMA诱导单核细胞株THP-1向巨噬细胞分化过程中,ACAT-1 mRNA、蛋白表达及酶活性水平明显增高(P<0.05),呈时间依赖效应。(3)巨噬细胞向泡沫细胞分化过程中,ACAT-1 mRNA、蛋白表达及酶活性水平增高,呈时间依赖效应。但和无血清培养巨噬细胞比较差异无统计学意义(P>0.05)。结论单核细胞株THP-1经PMA,PMA/ox-LDL诱导后,可分化为巨噬细胞、泡沫细胞。ACAT-1的表达及活性增高在单核细胞向泡沫细胞诱导分化过程中起着重要作用。
Objective To explore the expression of Acyl CoA cholesterol acyltrausferase-1 （ ACAT-1 ） in the cell differentiation from mononuclear cell to foam cell. Methods The mononuclear cell was resuscitated and cocultured with phorbol myristate acetate （PMA）, and then, the foam cell was induced with PMA plus oxidized low density lipoprotein （ox-LDL） for 1 -4 d. RT-PCR assay, western bloting assay were employed to examine the expression of ACAT-1 mRNA,protein expression in the cell differentiation. Results （ 1 ） The mononuclear cells line THP-1 could be differentiated into macrophages after 48 hours induction by the PMA,and into foam ceils after 24 hours induction by PMA/ox-LDL. （2）Compared with the mononuclear cell in the control group, expression of ACAT-1 mRNA and protein in macrophages obviously increased （ P 〈0.05 ） ,with a time dependant pattern. Compared with the macrophages, expression of ACAT-1 mRNA and protein, the ACAT-1 activity in foam cells increased,demonstrated time dependant pattern,without significant difference compared with the control group （ P 〉 0.05 ）. Conclusion The mononuclear cells line THP-1 could differentiate into macrophage and foam cells after induction by PMA and PMA/ox- LDL. The expression and activity of ACAT-1 gene increased may play an important role in the mononuclear cells differentiation into the foam cell.
出处
《疑难病杂志》
CAS
2009年第11期650-653,F0003,共5页
Chinese Journal of Difficult and Complicated Cases
