Child P, Kuksis A
Can J Biochem. 1980 Oct;58(10):1215-22. doi: 10.1139/o80-162.
Cellular uptake and transport of cholesterol and beta-sitosterol was studied in rat jejunal villus cells isolated by hyaluronidase dispersion. Sterol absorption was measured from micellar solutions using a Millipore filtration technique. In actively metabolizing cells both cholesterol and beta-sitosterol exhibited rapid initial uptake curves which leveled off after 10 min, corresponding approximately to a 1:1 sterol:phospholipid molar ratio within the microsomal membranes of the cell, which suggested a physicochemical end point. Using various proportions of cholesterol and beta-sitosterol in the incubation medium, it was shown that each sterol interfered with the absorption of the other, the one present in the highest concentration showing the relatively higher uptake. Mixtures of equimolar proportions were taken up by the cells in a ratio that slightly favoured beta-sitosterol, but significantly more cholesterol than beta-sitosterol was found to be transferred to the microsomes. The limited mass discrimination between cholesterol and beta-sitosterol seen in the isolated cells is in contrast to the preferential absorption of cholesterol in vivo. These studies demonstrate that the molecular basis for the differential transport of cholesterol and beta-sitosterol in the lymph observed in vivo must be sought at a higher level of physiological activity of the cells (e.g., chylomicron formation and secretion) than that achieved in the present experiments.
利用透明质酸酶分散法分离大鼠空肠绒毛细胞,研究胆固醇和β-谷甾醇的细胞摄取及转运。采用密理博过滤技术,从胶束溶液中测定甾醇吸收情况。在活跃代谢的细胞中,胆固醇和β-谷甾醇均呈现快速的初始摄取曲线,10分钟后趋于平稳,这大致对应于细胞微粒体膜内1:1的甾醇:磷脂摩尔比,表明达到了物理化学终点。在孵育培养基中使用不同比例的胆固醇和β-谷甾醇,结果显示每种甾醇都会干扰另一种甾醇的吸收,浓度最高的那种甾醇摄取相对较高。等摩尔比例的混合物被细胞摄取的比例略微有利于β-谷甾醇,但发现转移到微粒体中的胆固醇明显多于β-谷甾醇。在分离细胞中观察到的胆固醇和β-谷甾醇之间有限的质量区分与体内胆固醇的优先吸收形成对比。这些研究表明,体内观察到的淋巴中胆固醇和β-谷甾醇差异转运的分子基础,必须在比本实验所达到的更高细胞生理活性水平(例如乳糜微粒的形成和分泌)上寻找。