摘要
目的探讨白介素-1（IL-1）对单核细胞向泡沫细胞分化过程中脂酰辅酶A-胆固醇酰基转移酶-1（ACAT-1）蛋白表达和活性的影响。方法200nmol／L佛波酯（PMA）诱导THP-1单核细胞48h使其转化为巨噬细胞，流式细胞术检测CDl4的表达；巨噬细胞与80mg／L氧化型低密度脂蛋白（Ox-LDL）共孵育24h，使之向泡沫细胞分化，油红O染色观察细胞质内脂质沉积；Westernblot检测各组细胞（单核细胞组、巨噬细胞组、泡沫细胞组、泡沫细胞＋IL-1组、泡沫细胞＋IL-1／Anti-ILl组）ACAT-l的蛋白表达，液相闪烁计数法检测ACAT-1的酶活性。结果单核细胞株THP-1与200nM的PMA共孵育48h后，分化为巨噬细胞，CDld阳性表达率为85．7％；巨噬细胞与Ox-LDL共孵育24h后，油红O染色胞浆内可见大量吞噬的脂质小滴。与单核细胞组相比，巨噬细胞组、泡沫细胞组和泡沫细胞＋IL-1组ACAT-1蛋白表达上调，活性升高（P〈0．05），泡沫细胞＋IL-1／Anti-ILl组蛋白表达上调及活性升高不明显（P〉0．05）。结论IL-1对单核细胞向泡沫细胞分化过程中ACAT-1蛋白表达及酶活性有上调作用，IL-1单克隆抗体可以拮抗这种效应。
Objective Our current investigation attempts to explore the effects of IL-1 on the ACAT-1protein expression and activity in the mononuclear cells differentiation to foam cells. Methods The mononuclear cells （THP-1） were co-cultured with 200 nmol/L phorbol myristate acetate （PMA） and induced into differentiation to macrophages. Flow cytometry was employed to examine CD14 expression. The macrophages were cultured with 80 mg/L oxidized low density lipoprotein （Ox-LDL） and induced into differentiation to the foam cells, and oil red O staining was employed to examine cytoplasm lipid deposition. Western blotting assay and liquid phase scintillation counting were employed to examine the expression of ACAT-1 protein and ACAT-1 activity in different groups （the mononuclear cells group, the macrophages group, the foam cells group, the foam cells IL-1 group, the foam cells IL-1/IL-I monoclonal antibody group ）, respectively. Results The mononuclear cells were differentiated into maerophage cells after cultured in RPMI 1640 media containing 200 nM PMA for 48 h, and expressed the CD14 with an 85.7% positive rate. Some red stained substance could be found in the cytolymph after co-cultured with 80 mg/L Ox-LDL for 24 h. Compared with the mononuclear cells group, the protein expressionlevels and activity of ACAT-1 were found to markedly increase in the maerophages group, the loam cells group and the foam ceils IL-1 group （P〈0.05）. The protein expression levels and activity of ACAT-1 did not increase in the foam cells IL-1/IL-1 monoclonal antibody group （P〉0.05）. Conclusion IL-1 could up-regulate the expression of ACAT-1 protein arid the ACAT-1 activity during the mononuclear cells differentiation into the foam ceils, and IL-1 monoclonal antibody might resist the effect.
出处
《中国心血管病研究》
CAS
2011年第9期691-695,共5页
Chinese Journal of Cardiovascular Research
基金
山西省自然科学基金项目（2006011123）
作者简介
通讯作者：魏武，E-mail：tigergv@163．com
