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KH902 suppresses high glucose‐induced migration and sprouting of human retinal endothelial cells by blocking VEGF and PIGF
Author(s) -
Chen X.,
Li J.,
Li M.,
Zeng M.,
Li T.,
Xiao W.,
Li J.,
Wu Q.,
Ke X.,
Luo D.,
Tang S.,
Luo Y.
Publication year - 2013
Publication title -
diabetes, obesity and metabolism
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.445
H-Index - 128
eISSN - 1463-1326
pISSN - 1462-8902
DOI - 10.1111/dom.12008
Subject(s) - chemistry , western blot , vascular endothelial growth factor , kinase insert domain receptor , placental growth factor , microbiology and biotechnology , receptor , vascular endothelial growth factor a , cancer research , medicine , vegf receptors , biology , biochemistry , gene
Aims Vascular endothelial growth factor ( VEGF ) and placental growth factor ( PlGF ) are upregulated in many ocular neovascular diseases such as diabetic retinopathy ( DR ). KH902 is a recombinant fusion protein with its binding ligand taken from the domains of VEGF receptor‐1 ( VEGFR ‐1) and VEGF receptor‐2 ( VEGFR ‐2) and can bind all VEGF ‐A isoforms and PlGF . The aim of this study was to investigate the underlying mechanisms of anti‐angiogenic effects of KH902 . Methods The toxic effect of KH902 on cultured human retinal endothelial cells ( HRECs ) was measured by Annexin V/ PI staining and MTT assay. The concentrations of secreted VEGF and PlGF were measured by ELISA . The migration of HRECs was assessed by scratch wound and transwell assay. The sprouting of HRECs was determined by tube formation assay. The protein levels of Src, p‐Src, PI3K , Akt1, p‐Akt1, Erk1/2 and p‐Erk1/2 were measured by Western blot. Results KH902 at the concentrations from 100 ng/ml to 100 µg/ml had no cytotoxicity to cultured HRECs . KH902 bound not only VEGF165 , but also PlGF that were secreted by HRECs under high glucose condition. A 500 ng/ml of KH902 significantly suppressed high glucose‐induced migration and sprouting of HRECs through downregulating the expression of PI3K and inhibiting the activation of Src, Akt1 and Erk1/2. Conclusion Our study indicates that KH902 suppresses high glucose‐induced migration and sprouting of HRECs through not only binding VEGF , but also PlGF to inhibit the activation of Src‐Akt1‐Erk1/2 pathway. KH902 is a drug that potentially inhibits angiogenic pathways involving in DR or other ocular neovascular diseases.