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培养的脑、肾上腺和主动脉内皮细胞葡萄糖转运的调节。

Modulation of cultured brain, adrenal, and aortic endothelial cell glucose transport.

作者信息

Gaposchkin C G, Garcia-Diaz J F

机构信息

Department of Physiology, Boston University School of Medicine, MA 02118, USA.

出版信息

Biochim Biophys Acta. 1996 Dec 4;1285(2):255-66. doi: 10.1016/s0005-2736(96)00172-1.

Abstract

Studies of glucose transporter activity and anti-glucose transporter (GLUT1) immunoblots were performed on different endothelial cell primary cultures (brain capillary, adrenal capillary and aortic) to determine their response to glucose deprivation. Cell cultures were exposed to glucose deprivation (0.5 mM) for 48 h periods and refed (11.0 mM) for 36 additional hours. Control cultures were kept in 11.0 mM glucose for the duration of these studies. Measurements of 2-[3H]deoxy-D-glucose uptake and membrane fraction purification were performed every 12 h during these timecourses. Baseline cytochalasin-B sensitive uptake of 2-deoxy-D-glucose was near three times larger in brain capillary endothelial cells than in adrenal or aortic endothelial cultures. In all three endothelial cell cultures, 2-deoxy-D-glucose uptake increased during glucose deprivation, and returned to control values upon refeeding. Aortic and adrenal cortical endothelia expressed the starvation induced increases 12 h sooner than brain capillary endothelia. Return to control values was also 12 h faster in these cultured endothelia. Immunoblot studies showed that in all three endothelial cell cultures the increases in transporter activity during glucose starvation correlate with increased membrane expression of GLUT1. Quantitative analysis of the anti-GLUT1 immunoblots indicated that induction of GLUT1 following glucose starvation was slower in brain capillary endothelia than in aortic or adrenal endothelia. The slower response by brain capillary endothelial cells may be related to the higher transport rate of glucose in these cells.

摘要

对不同的内皮细胞原代培养物(脑毛细血管、肾上腺毛细血管和主动脉)进行葡萄糖转运蛋白活性研究和抗葡萄糖转运蛋白(GLUT1)免疫印迹分析,以确定它们对葡萄糖剥夺的反应。将细胞培养物暴露于葡萄糖剥夺(0.5 mM)环境中48小时,然后再补充葡萄糖(11.0 mM)36小时。在这些研究过程中,对照培养物始终保持在11.0 mM葡萄糖环境中。在这些时间进程中,每12小时进行一次2-[3H]脱氧-D-葡萄糖摄取量测量和膜组分纯化。脑毛细血管内皮细胞中,细胞松弛素B敏感的2-脱氧-D-葡萄糖摄取基线比肾上腺或主动脉内皮细胞培养物中的高近三倍。在所有三种内皮细胞培养物中,葡萄糖剥夺期间2-脱氧-D-葡萄糖摄取增加,再喂食后恢复到对照值。主动脉和肾上腺皮质内皮细胞饥饿诱导的增加比脑毛细血管内皮细胞早12小时出现。在这些培养的内皮细胞中,恢复到对照值的速度也快12小时。免疫印迹研究表明,在所有三种内皮细胞培养物中,葡萄糖饥饿期间转运蛋白活性的增加与GLUT1膜表达的增加相关。抗GLUT1免疫印迹的定量分析表明,葡萄糖饥饿后脑毛细血管内皮细胞中GLUT1的诱导比主动脉或肾上腺内皮细胞慢。脑毛细血管内皮细胞的较慢反应可能与这些细胞中较高的葡萄糖转运速率有关。

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