Blockade by phosphorothioate aptamers of advanced glycation end products-induced damage in cultured pericytes and endothelial cells

Yuichiro Higashimoto, Takanori Matsui, Yuri Nishino, Junichi Taira, Hiroyoshi Inoue, Masayoshi Takeuchi, Sho ichi Yamagishi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Advanced glycation end products (AGEs) not only inhibit DNA synthesis of retinal pericytes, but also elicit vascular hyperpermeability, pathological angiogenesis, and thrombogenic reactions by inducing vascular endothelial growth factor (VEGF) and plasminogen activator inhibitor-1 (PAI-1) through the interaction with the receptor for AGEs (RAGE), thereby being involved in the pathogenesis of diabetic retinopathy. In this study, we screened novel phosphorothioate-modified aptamers directed against AGEs (AGEs-thioaptamers) using a combinatorial chemistry in vitro, and examined whether these aptamers could inhibit the AGE-induced damage in both retinal pericytes and human umbilical vein endothelial cells (HUVECs). We identified 11 AGEs-thioaptamers; among them, clones #4, #7s and #9s aptamers had higher binding affinity to AGEs-human serum albumin (HSA) than the others. Surface plasmon resonance analysis revealed that KD values of #4s, #7s and #9s were 0.63, 0.36, and 0.57nM, respectively. Furthermore, these 3 clones dose-dependently restored the decrease in DNA synthesis in AGE-exposed pericytes. AGEs significantly increased RAGE, VEGF and PAI-1 mRNA levels in HUVEC, all of which were completely blocked by the treatment with 20nM clone #4s aptamer. Quartz crystal microbalance analysis confirmed that #4s aptamer dose-dependently inhibited the binding of AGEs-HSA to RAGE. Our present study demonstrated that AGEs-thioaptamers could inhibit the harmful effects of AGEs in pericytes and HUVEC by suppressing the binding of AGEs to RAGE. Blockade by AGEs-thioaptamers of the AGEs-RAGE axis might be a novel therapeutic strategy for diabetic retinopathy.

Original languageEnglish
Pages (from-to)64-70
Number of pages7
JournalMicrovascular Research
Volume90
DOIs
Publication statusPublished - 2013 Nov

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Pericytes
Advanced Glycosylation End Products
Endothelial cells
Cultured Cells
Human Umbilical Vein Endothelial Cells
Endothelial Cells
Clone Cells
Plasminogen Activator Inhibitor 1
Diabetic Retinopathy
Serum Albumin
Albumin Receptors
Vascular Endothelial Growth Factor A
Quartz Crystal Microbalance Techniques
Pathologic Neovascularization
Vascular Endothelial Growth Factor Receptor
Surface Plasmon Resonance
DNA
Quartz crystal microbalances
Blood Vessels
Surface plasmon resonance

ASJC Scopus subject areas

  • Biochemistry
  • Cardiology and Cardiovascular Medicine
  • Cell Biology

Cite this

Blockade by phosphorothioate aptamers of advanced glycation end products-induced damage in cultured pericytes and endothelial cells. / Higashimoto, Yuichiro; Matsui, Takanori; Nishino, Yuri; Taira, Junichi; Inoue, Hiroyoshi; Takeuchi, Masayoshi; Yamagishi, Sho ichi.

In: Microvascular Research, Vol. 90, 11.2013, p. 64-70.

Research output: Contribution to journalArticle

Higashimoto, Yuichiro ; Matsui, Takanori ; Nishino, Yuri ; Taira, Junichi ; Inoue, Hiroyoshi ; Takeuchi, Masayoshi ; Yamagishi, Sho ichi. / Blockade by phosphorothioate aptamers of advanced glycation end products-induced damage in cultured pericytes and endothelial cells. In: Microvascular Research. 2013 ; Vol. 90. pp. 64-70.
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