title: Inhibition of high glucose-induced VEGF and ICAM-1 expression in human retinal pigment epithelium cells by targeting ILK with small interference RNA.
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.
authors.affiliation: Ophthalmology Hospital, First Affiliated Hospital, Harbin Medical University, 23 Youzheng Street, Harbin, 150001, Heilongjiang Province, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Makoto
authors.initials: M
authors.lastname: Matsukura
authors.forename: Isao
authors.initials: I
authors.lastname: Fujii
authors.forename: Kaori
authors.initials: K
authors.lastname: Ito
authors.forename: Ji En
authors.initials: JE
authors.lastname: Zhao
authors.forename: Makoto
authors.initials: M
authors.lastname: Shinohara
authors.forename: Ye Qing
authors.initials: YQ
authors.lastname: Wang
authors.forename: Xiao Mei
authors.initials: XM
authors.lastname: Zhang
mesh_terms: D000704:Analysis of Variance; D015153:Blotting, Western; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D055786:Gene Knockout Techniques; D005947:Glucose / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000235:genetics; D055213:Retinal Pigment Epithelium / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*; D000099200:Scaffold Protein ILK
chemical_list: D017931:DNA Primers; D005947:Glucose; D018799:Intercellular Adhesion Molecule-1; D017346:Protein Serine-Threonine Kinases; D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A; D000099200:Scaffold Protein ILK; C467484:VEGFA protein, human