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Lutein transport by Caco-2 TC-7 cells occurs partly by a facilitated process involving the scavenger receptor class B type I (SR-BI)

机译:叶绿素通过Caco-2 TC-7细胞的运输部分通过促进过程进行该过程涉及I类清道夫受体(SR-BI)

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摘要

The carotenoid lutein is thought to play a role in the human eye and to protect against age-related macular degeneration. Lutein transport in the human intestine has not been characterized. We examined lutein transport processes using Caco-2 TC-7 monolayers as a model for human intestinal epithelium. Purified lutein was mixed with phospholipids, lysophospholipids, cholesterol, mono-olein, oleic acid and taurocholate to obtain lutein-rich mixed micelles that mimicked those found under physiological conditions. The micelles were added to the apical side of Caco-2 TC-7 cell monolayers for 30 min or 3 h at 37 °C. Absorbed lutein, i.e. the sum of lutein recovered in the scraped cells and in the basolateral chamber, was quantified by HPLC. Transport rate was measured (i) as a function of time (from 15 to 60 min), (ii) as a function of micellar lutein concentration (from 1.5 to 15 μM), (iii) at 4 °C, (iv) in the basolateral to apical direction, (v) after trypsin pretreatment, (vi) in the presence of β-carotene and/or lycopene, (vii) in the presence of increasing concentrations of antibody against SR-BI (scavenger receptor class B type 1) and (viii) in the presence of increasing concentrations of a chemical inhibitor of the selective transfer of lipids mediated by SR-BI, i.e. BLT1 (blocks lipid transport 1). The rate of transport of lutein as a function of time and as a function of concentration was saturable. It was significantly lower at 4 °C than at 37 °C (approx. 50%), in the basal to apical direction than in the opposite direction (approx. 85%), and after trypsin pretreatment (up to 45%). Co-incubation with β-carotene, but not lycopene, decreased the lutein absorption rate (approx. 20%) significantly. Anti-SR-BI antibody and BLT1 significantly impaired the absorption rate (approx. 30% and 57% respectively). Overall, these results indicate that lutein absorption is, at least partly, protein-mediated and that some lutein is taken up through SR-BI.
机译:类胡萝卜素叶黄素被认为在人眼中起着一定的作用,并能防止与年龄有关的黄斑变性。叶黄素在人肠中的运输尚未鉴定。我们检查叶黄素运输过程中使用Caco-2 TC-7单层作为人肠上皮的模型。将纯化的叶黄素与磷脂,溶血磷脂,胆固醇,单油精,油酸和牛磺胆酸盐混合,以获得富含叶黄素的混合胶束,该胶束模仿在生理条件下发现的胶束。将胶束在37°C下添加到Caco-2 TC-7细胞单层的顶端30分钟或3小时。通过HPLC定量吸收的叶黄素,即在刮擦的细胞和基底外侧室中回收的叶黄素的总和。 (i)时间(15至60分钟)的函数,(ii)胶束叶黄素浓度(1.5至15μM)的函数,(iii)在4°C下,(iv) (vi)胰蛋白酶预处理后,(vi)在β-胡萝卜素和/或番茄红素的存在下,(vii)在抗SR-BI(B类清道夫受体)抗体浓度增加的情况下, )和(viii)在由SR-BI介导的脂质(即BLT1)介导的脂质选择性转移的化学抑制剂浓度增加的情况下(阻断脂质转运1)。叶黄素的运输速率随时间和浓度的变化是饱和的。在胰蛋白酶预处理后(45%),在4°C下比在37°C下(约50%),从基础到顶部的方向显着低于相反的方向(约85%)。与β-胡萝卜素而非番茄红素共同孵育可显着降低叶黄素吸收率(约20%)。抗SR-BI抗体和BLT1大大削弱了吸收率(分别约为30%和57%)。总体而言,这些结果表明叶黄素的吸收至少部分是蛋白质介导的,并且某些叶黄素是通过SR-BI吸收的。

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