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用小干扰 RNA 靶向整合素连接激酶抑制高糖诱导的人视网膜色素上皮细胞血管内皮生长因子和细胞间黏附分子-1 的表达。

Inhibition of high glucose-induced VEGF and ICAM-1 expression in human retinal pigment epithelium cells by targeting ILK with small interference RNA.

机构信息

Ophthalmology Hospital, First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Harbin, 150001, Heilongjiang Province, China.

出版信息

Mol Biol Rep. 2012 Jan;39(1):613-20. doi: 10.1007/s11033-011-0778-2. Epub 2011 May 7.

Abstract

The aim of this study was to investigate the change of Integrin-linked kinase (ILK) expression of human retinal pigment epithelium (RPE) cells in response to high glucose, and the effect of targeting ILK with small interference RNA (siRNA) on the high glucose-induced expression of vascular endothelial growth factor (VEGF) and intercellular adhesion molecule-1 (ICAM-1). The ILK mRNA and protein expression in human RPE cells were analyzed with RT-PCR and western blot after exposure to 5.5, 30, 40, 50 mM glucose, or 5.5 mM glucose+45.5 mM mannitol for 48 h. The expression of VEGF and ICAM-1 was also determined. Cells were treated with ILK siRNA, to determine the effect of ILK on VEGF and ICAM-1 expression following treatment with high glucose. High concentrations of glucose significantly up-regulated ILK mRNA and protein expression, and the ILK expression increased along with the glucose concentration. The changes of VEGF and ICAM-1 expression were similar to that of ILK expression. Knocking down ILK gene expression with siRNA inhibited the elevation of VEGF and ICAM-1 induced by high glucose treatment. These results suggested that ILK was involved in the response of RPE cells to high glucose and may therefore play a role in the pathogenesis of diabetic ophthalmology.

摘要

本研究旨在探讨整合素连接激酶(ILK)在人视网膜色素上皮(RPE)细胞对高糖反应中的表达变化,以及用小干扰 RNA(siRNA)靶向 ILK 对高糖诱导的血管内皮生长因子(VEGF)和细胞间黏附分子-1(ICAM-1)表达的影响。在将人 RPE 细胞暴露于 5.5、30、40、50 mM 葡萄糖或 5.5 mM 葡萄糖+45.5 mM 甘露醇 48 小时后,通过 RT-PCR 和 Western blot 分析 ILK mRNA 和蛋白表达。还测定了 VEGF 和 ICAM-1 的表达。用 ILK siRNA 处理细胞,以确定在高糖处理后 ILK 对 VEGF 和 ICAM-1 表达的影响。高浓度葡萄糖显著上调 ILK mRNA 和蛋白表达,并且随着葡萄糖浓度的增加,ILK 表达增加。VEGF 和 ICAM-1 表达的变化与 ILK 表达的变化相似。用 siRNA 敲低 ILK 基因表达抑制了高糖处理诱导的 VEGF 和 ICAM-1 的升高。这些结果表明,ILK 参与了 RPE 细胞对高糖的反应,因此可能在糖尿病眼病的发病机制中起作用。

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