Use of cyclodextrins for manipulating cellular cholesterol content.
Use of cyclodextrins for manipulating cellular cholesterol content.
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DOI：
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发表时间：
1997-11
影响因子：
6.5
通讯作者：
A. E. Christian;M. Haynes;M. Phillips;G. Rothblat
中科院分区：
文献类型：
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作者：
A. E. Christian;M. Haynes;M. Phillips;G. Rothblat
该实验室以前的研究表明，组织培养细胞暴露于环糊精导致胆固醇快速消耗。在本研究中，我们已经开发了实验系统，用于使用环糊精的溶液，无论是2-羟丙基β-环糊精或甲基化β-环糊精，与不同量的游离胆固醇复合，以操纵细胞胆固醇含量。已发现通过环糊精递送的胆固醇具有代谢活性，如通过酰基辅酶A：胆固醇酰基转移酶（ACAT）介导的[3 H]胆固醇在Fu 5AH大鼠肝癌细胞和中国仓鼠卵巢细胞中的酯化所测量的。在所有研究的细胞类型中，甲基化β-环糊精被发现是更有效的供体，6小时内平均胆固醇摄取至少为100微克胆固醇/毫克蛋白质。通过改变环糊精：胆固醇的摩尔比，可以操纵细胞的细胞胆固醇含量，产生从净胆固醇富集到耗尽的条件。环糊精的使用为调节组织培养细胞的胆固醇含量提供了方便、精确和可重复的方法。
Previous studies from this laboratory have demonstrated that exposure of tissue culture cells to cyclodextrins results in rapid cholesterol depletion. In the present study, we have developed experimental systems for using solutions of cyclodextrins, either 2-hydroxypropyl beta-cyclodextrin or methylated beta-cyclodextrin, complexed with varying amounts of free cholesterol to manipulate cell cholesterol content. Cholesterol delivered via the cyclodextrin has been found to be metabolically active, as measured by the acyl-coenzyme A:cholesterol acyltransferase (ACAT)-mediated esterification of [3H]cholesterol in Fu5AH rat hepatoma cells and Chinese hamster ovary cells. The methylated beta-cyclodextrin was found to be a more efficient donor in all cell types studied, with an average cholesterol uptake of at least 100 microg cholesterol/mg protein within 6 h. By modifying the cyclodextrin:cholesterol molar ratio, it is possible to manipulate the cellular cholesterol content of cells, producing conditions ranging from net cholesterol enrichment to depletion. The use of cyclodextrins provides a convenient, precise and reproducible method for modulating the cholesterol content of tissue culture cells.