Antisense GLUT-1 protects mesangial cells from glucose induction of GLUT-1 and fibronectin expression
Antisense GLUT-1 protects mesangial cells from glucose induction of GLUT-1 and fibronectin expression
复制标题
DOI：
10.1152/ajprenal.2001.280.4.f657
复制
发表时间：
2001-04-01
影响因子：
4.2
通讯作者：
Brosius, FC III
中科院分区：
文献类型：
--
作者：
Heilig, CW;Kreisberg, JI;Brosius, FC III
建立了一个稳定表达反义GLUT-1的大鼠肾小球系膜细胞克隆(即MCGT1AS细胞)，以保护其免受高糖暴露。MCGT1AS组GLUT-1蛋白减少50%，2-脱氧-[H-3]葡萄糖摄取率降低33%。以MCLacZ对照细胞和MCGT1 GLUT-1过表达细胞为对照。在MCLacZ中，20 mM D-葡萄糖使GLUT-1转录增加了90%，而在MCGT1AS中没有增加。葡萄糖(8 MM)和12 mM木糖醇[一种己糖一磷酸(HMP)的分流底物]不能刺激GLUT-1转录。用3-O-甲基葡萄糖取代标准的8 mM D-葡萄糖87%可使Glut-1转录减少80%。与对照组相比，D-葡萄糖(20 MM)可使MCLacZ和MCGT1AS的纤维连接蛋白mRNA和蛋白分别增加47%和100%。纤维连接蛋白合成在MCGT1组增加48%，在MCGT1AS组减少44%。我们得出结论：1)GLUT-1对D-葡萄糖的反应依赖于葡萄糖代谢，但不是通过HMP分流；2)反义GLUT-1处理肾小球系膜细胞可以阻断D-葡萄糖诱导的GLUT-1和纤维连接蛋白的表达，从而证明了一种保护作用，这可能有利于糖尿病的发生。
A stable clone of rat mesangial cells expressing antisense GLUT-1 (i.e., MCGT1AS cells) was developed to protect them from high glucose exposure. GLUT-1 protein was reduced 50%, and the 2-deoxy-[H-3] glucose uptake rate was reduced 33% in MCGT1AS. MCLacZ control cells and MCGT1 GLUT-1-overexpressing cells were used for comparisons. In MCLacZ, 20 mM D-glucose increased GLUT-1 transcription 90% vs. no increase in MCGT1AS. Glucose (8 mM) and 12 mM xylitol [a hexose monophosphate (HMP) shunt substrate] did not stimulate GLUT-1 transcription. An 87% replacement of the standard 8 mM D-glucose with 3-O-methylglucose reduced GLUT-1 transcription 80%. D-Glucose (20 mM) increased fibronectin mRNA and protein by 47 and 100%, respectively, in MCLacZ vs. no increases in MCGT1AS. Fibronectin synthesis was elevated 48% in MCGT1 and reduced 44% in MCGT1AS. We conclude that 1) transcription of GLUT-1 in response to D-glucose depends on glucose metabolism, although not through the HMP shunt, and 2) antisense GLUT-1 treatment of mesangial cells blocks D-glucose-induced GLUT-1 and fibronectin expression, thereby demonstrating a protective effect that could be beneficial in the setting of diabetes.