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16 September 2019 : Laboratory Research  

The Effects of the CXCR4 Antagonist, AMD3465, on Human Retinal Vascular Endothelial Cells (hRVECs) in a High Glucose Model of Diabetic Retinopathy

Di Wu1ABE*, Li Jin1CF, Hongshuang Xu1B

DOI: 10.12659/MSM.917186

Med Sci Monit 2019; 25:6946-6954

Abstract

BACKGROUND: High blood glucose levels in diabetes result in retinal angiogenesis, which is the key feature of diabetic retinopathy. This study aimed to investigate the effects of the CXCR4 antagonist, AMD3465, on human retinal vascular endothelial cells (hRVECs) in vitro.

MATERIAL AND METHODS: Cell viability and the protein expression levels of CXCR4 and stromal cell-derived factor 1 (SDF-1) were evaluated in high glucose (HG)-treated human retinal vascular endothelial cells (hRVECs). The cell counting kit 8 (CCK-8) assay, the colony formation assay, immunofluorescence, and Western blot were used to investigate the effects of AMD3465 on hRVEC cell viability, colony formation, cell proliferation, and expression of CXCR4 and SDF-1. Cell apoptosis and angiogenesis were assessed by flow cytometry and Western blot.

RESULTS: Treatment with high glucose reduced the viability of hRVECs and increased the protein expression levels of CXCR4 and SDF-1. Following treatment with AMD3465, the colony formation capacity and cell proliferation in hRVECs increased, and there was a significant reduction in apoptosis rate compared with the untreated cells. AMD3465 significantly reduced the expression of angiogenesis-associated proteins, including ICAM1, VCAM1, VEGF, and AngII. AMD3465 significantly reduced the protein expression levels of TNF-α, IL-1β, NF-κB, and p-p65.

CONCLUSIONS: The CXCR4 antagonist, AMD3465, reduced apoptosis of HG-treated hRVECs in an in vitro model of diabetic retinopathy.

Keywords: Diabetic Retinopathy, Protective Agents, Receptors, CXCR4, Apoptosis, Cell Survival, Chemokine CXCL12, endothelial cells, Glucose, Intercellular Adhesion Molecule-1, Neovascularization, Pathologic, Retina, Retinal Vessels, Vascular Endothelial Growth Factor A

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750