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葡萄糖转运蛋白1在角膜上皮伤口修复过程中表达增强。

Glucose transporter 1 expression is enhanced during corneal epithelial wound repair.

作者信息

Takahashi H, Kaminski A E, Zieske J D

机构信息

Schepens Eye Research Institute, Boston, MA, USA.

出版信息

Exp Eye Res. 1996 Dec;63(6):649-59. doi: 10.1006/exer.1996.0159.

Abstract

Corneal epithelial wound healing involves a number of metabolically demanding processes such as cell migration and proliferation. The energy for those processes is known to be provided by glycolysis. Thus, it was hypothesized that migrating epithelium would require high levels of glucose to provide a substrate for glycolysis. It is well established that glucose is transported into virtually all mammalian cells by the facilitated glucose transport protein, glucose transporter (GLUT). We sought to investigate the expression of one of the isoforms of glucose transporter, GLUT1, in corneal epithelium after epithelial debridement in the rat. Three-millimeter debridement wounds were made on central rat cornea and allowed to heal from 1 hr to 21 days. To quantitate changes in GLUT1 mRNA and protein levels, whole corneal epithelium was harvested and analysed by reverse transcription-polymerase chain reaction, northern blot, and western blot analysis. GLUT1 protein localization was also observed immunohistochemically. Expression of GLUT1 protein rapidly increased following wounding and was 2.4-fold higher than control at 4 hr post debridement. GLUT1 protein levels continued to increase even after epithelial wound closure (24 hr) and peaked at 2 days post debridement, 5.8-fold higher than control. The increase in GLUT1 protein levels coincided with enhanced GLUT1 mRNA levels (3.7-fold higher than control at 4 hr post debridement). Immunofluorescence microscopy showed increased binding of anti-GLUT1 concentrated in the membranes of the basal cells from limbus-to-limbus until 24 hr post wounding. After 24 hr. binding remained enhanced in the wound area, while binding to the limbal basal cells returned to the control level. In conclusion, the expression of GLUT1 mRNA and protein are rapidly enhanced after wounding. This may allow the increased transport of glucose, providing the metabolic energy necessary for cell migration and proliferation.

摘要

角膜上皮伤口愈合涉及许多对代谢要求较高的过程,如细胞迁移和增殖。已知这些过程的能量由糖酵解提供。因此,有人推测迁移的上皮细胞需要高水平的葡萄糖来为糖酵解提供底物。众所周知,葡萄糖通过易化葡萄糖转运蛋白——葡萄糖转运体(GLUT)进入几乎所有哺乳动物细胞。我们试图研究大鼠角膜上皮清创术后葡萄糖转运体的一种异构体GLUT1的表达情况。在大鼠角膜中央制作3毫米的清创伤口,让其从1小时到21天愈合。为了定量GLUT1 mRNA和蛋白质水平的变化,收集整个角膜上皮,通过逆转录-聚合酶链反应、Northern印迹和Western印迹分析进行检测。还通过免疫组织化学观察GLUT1蛋白的定位。伤口形成后,GLUT1蛋白的表达迅速增加,清创后4小时比对照高2.4倍。即使上皮伤口闭合(24小时)后,GLUT1蛋白水平仍持续升高,并在清创后2天达到峰值,比对照高5.8倍。GLUT1蛋白水平的增加与GLUT1 mRNA水平的升高相一致(清创后4小时比对照高3.7倍)。免疫荧光显微镜显示,抗GLUT1的结合增加,在伤口形成后24小时内集中在从角膜缘到角膜缘的基底细胞膜上。24小时后。伤口区域的结合仍然增强,而角膜缘基底细胞的结合恢复到对照水平。总之,受伤后GLUT1 mRNA和蛋白的表达迅速增强。这可能会增加葡萄糖的转运,为细胞迁移和增殖提供必要的代谢能量。

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