【摘要】 目的观察不对称二甲基精氨酸(ADMA)对单核/巨噬细胞分化为泡沫细胞过程中酰基辅酶A胆固醇酰基转移酶-1(ACAT-1)表达的影响。方法 1)THP-1单核细胞与160 nmol/L佛波酯(PMA)孵育48 h后,再与80 mg/L氧化型低密度脂蛋白(ox-LDL)孵育24 h,用油红O染色在光镜下鉴定细胞形态及变化。2)将THP-1单核细胞、巨噬细胞、泡沫细胞分别用20μmol/L ADMA干预24 h,酶法测泡沫细胞内胆固醇酯的含量,RT-PCR方法检测ACAT-1mRNA表达,Western blot方法检测其蛋白表达。结果 1)ADMA可显著增加泡沫细胞内胆固醇酯的含量。2)与THP-1单核细胞相比,巨噬细胞(P<0.05)、泡沫细胞(P<0.01)中ACAT-1 mRNA及蛋白表达增加;而与巨噬细胞相比,泡沫细胞中ACAT-1 mRNA及蛋白表达增加但无显著性差异。3)与各自的对照组相比,经ADMA作用后3种细胞中ACAT-1 mRNA及蛋白表达水平均增加(P<0.01)。结论 ADMA增加泡沫细胞内胆固醇酯的含量可能是通过上调ACAT-1的mRNA及蛋白表达来实现的。更多还原

【Abstract】 Objective To investigate the effects of Asymmetric dimethylarginine(ADMA) on the expression of acyl-coenzyme A:acylcholesterol transferase-1(ACAT-1) in cell differentiation from mononuclear/macrophage cells to foam cells.Methods 1) Cultured THP-1 monocyte cell was induced into macrophages by 160 nmol/L PMA for 48 h and then incubated with 80 mg/L ox-LDL for 24 h macrophages were induced to foam cells.2)After 20 μmol/L ADMA was incubated with THP-1 monocyte cells,macrophage cells and foam cells for 24 h,the cholesterolester content of foam cells was measured by enzyme assay.The expression of ACAT-1 in mRNA and protein was measured by RT-PCR and Western blot respectively.Results 1)ADMA increased the cholesterolester content of foam cells.2)Compared with the monocyte cells,expression of ACAT-1 in mRNA and protein in macrophage cells(P<0.05) and foam cells(P<0.01) obviously increased.The expression of ACAT-1 in mRNA and proteinin foam cells increased without significant difference.3)Compared with the control group respectively treatment of monocyte cells,macrophage cells and foam cells with ADMA result in an increase in mRNA and protein levels for ACAT-1(P<0.01).Conclusions ADMA may increase the cholesterol content of foam cells via upregulating the expression of ACAT-1 in the mRNA and protein.更多还原