Hassall D G, Graham A
Wellcome Research Laboratories, Kent, United Kingdom.
Cytometry. 1995 Dec 1;21(4):352-62. doi: 10.1002/cyto.990210407.
The free cholesterol content of cells can be monitored by the intensity of fluorescence emissions from the polyene antibiotic filipin. In a previous study (Hassall: Cytometry 13:381-388, 1992) using THP-1 macrophages, a decrease in filipin fluorescence in response to increasing concentrations of modified lipoprotein was observed, suggesting a reduction in the free cholesterol content of the cells. In this study, THP-1 macrophages were treated with a number of agents known to modulate cholesterol biosynthesis and cholesterol esterification. Changes in filipin fluorescence emissions were measured by flow cytometry, and correlated with changes in cholesterol biosynthesis measured by incorporation of [14C]acetate into cholesterol. A correlation between decreases in filipin fluorescence and reductions in cholesterol biosynthesis was apparent, even when cholesterol esterification was inhibited. These results suggest that the decreases in filipin fluorescence observed may be due, at least in part, to reduction in cholesterol biosynthesis.
细胞中的游离胆固醇含量可通过多烯抗生素制霉菌素的荧光发射强度来监测。在先前一项使用THP-1巨噬细胞的研究中（哈索尔：《细胞计数》13:381 - 388，1992年），观察到随着修饰脂蛋白浓度增加，制霉菌素荧光减弱，这表明细胞中游离胆固醇含量降低。在本研究中，用多种已知可调节胆固醇生物合成和胆固醇酯化的试剂处理THP-1巨噬细胞。通过流式细胞术测量制霉菌素荧光发射的变化，并与通过将[14C]乙酸盐掺入胆固醇来测量的胆固醇生物合成变化相关联。即使胆固醇酯化受到抑制，制霉菌素荧光降低与胆固醇生物合成减少之间的相关性也很明显。这些结果表明，观察到的制霉菌素荧光降低可能至少部分归因于胆固醇生物合成的减少。
