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发育中兔回肠刷状缘膜囊泡对牛磺胆酸盐的转运:结构/功能关系

Taurocholate transport by brush-border membrane vesicles from the developing rabbit ileum: structure/function relationships.

作者信息

Schwarz S M, Watkins J B, Ling S C

机构信息

Department of Pediatrics, New York Medical College, Valhalla 10595.

出版信息

J Pediatr Gastroenterol Nutr. 1990 May;10(4):482-9. doi: 10.1097/00005176-199005000-00012.

Abstract

To examine the ontogenesis of bile acid transport in the rabbit ileum, brush-border membrane vesicles (12- to 20-fold purified) were prepared from 14- to 49-day-old animals. Taurocholate uptake was characterized by the emergence of secondary active, Na(+)-dependent transport at the start of weaning (21 days). Transient intravesicular accumulation (overshoot) of taurocholate occurred at 5-10 s of incubation, and the overshoot maximum increased significantly from 21 days (349.2 +/- 22.4 nmol/mg protein) to 35 days (569.0 +/- 84.3 nmol/mg protein; p less than 0.001), without further increase at maturity (49 days, not equal to 607.6 +/- 136.7 nmol/mg protein). No significant taurocholate active uptake component was noted at 14 days; however, ileal vesicles from sucklings showed carrier-mediated, Na+ D-glucose cotransport. In greater than or equal to 35-day-old rabbits, osmolarity studies at 20 s of incubation showed that only approximately 12% of [14C]taurocholate uptake was secondary to bile acid-to-membrane binding. Conversely, at 20 min, greater than 95% of radiolabel incorporation represented solute bound to the external and/or internal membrane surface. Arrhenius plots establish brush-border membrane taurocholate uptake as an intrinsic, lipid-dependent process, with a slope discontinuity between 24 and 28 degrees C, similar to the membrane lipid thermotropic transition region. Steady-state fluorescence polarization studies (1,6-diphenyl-1,3,5-hexatriene) demonstrate a temporal association between the maturation of taurocholate uptake and age-related decreases in ileal brush-border membrane fluidity. These data indicate that maturation of bile acid secondary active transport in the rabbit ileum may be regulated, at least in part, by changes in brush-border membrane lipid dynamics.

摘要

为研究兔回肠中胆汁酸转运的个体发生过程,从14至49日龄的动物制备了刷状缘膜囊泡(纯化了12至20倍)。牛磺胆酸盐摄取的特征是在断奶开始时(21日龄)出现继发性主动、依赖Na⁺的转运。牛磺胆酸盐在孵育5至10秒时出现短暂的囊泡内积累(过冲),过冲最大值从21日龄时的(349.2±22.4 nmol/mg蛋白质)显著增加至35日龄时的(569.0±84.3 nmol/mg蛋白质;p<0.001),在成熟时(49日龄,607.6±136.7 nmol/mg蛋白质)未进一步增加。14日龄时未观察到明显的牛磺胆酸盐主动摄取成分;然而,乳兔的回肠囊泡显示出载体介导的Na⁺-D-葡萄糖共转运。在≥35日龄的兔子中,孵育20秒时的渗透压研究表明,[¹⁴C]牛磺胆酸盐摄取中只有约12%继发于胆汁酸与膜的结合。相反,在20分钟时,超过95%的放射性标记掺入代表与外膜和/或内膜表面结合的溶质。阿累尼乌斯图表明刷状缘膜牛磺胆酸盐摄取是一个内在的、依赖脂质的过程,在24至28℃之间斜率出现间断,类似于膜脂质热致转变区域。稳态荧光偏振研究(1,6-二苯基-1,3,5-己三烯)表明牛磺胆酸盐摄取的成熟与回肠刷状缘膜流动性随年龄相关的降低之间存在时间关联。这些数据表明,兔回肠中胆汁酸继发性主动转运的成熟可能至少部分受刷状缘膜脂质动力学变化的调节。

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