Activation of the VEGF-A/ERK/PLA2 axis mediates early retinal endothelial cell damage induced by high glucose: New insight from an in vitro model of diabetic retinopathy
| Autor: | Filippo Drago, Debora Lo Furno, Salvatore Salomone, Federica Conti, Florinda Gennuso, Giuliana Mannino, Gabriella Lupo, Carmelina Daniela Anfuso, Giovanni Giurdanella, Claudio Bucolo |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
MAPK/ERK pathway
Vascular Endothelial Growth Factor A Angiogenesis Inhibitors VEGF-A lcsh:Chemistry chemistry.chemical_compound Phospholipase A2 Receptors Extracellular Signal-Regulated MAP Kinases lcsh:QH301-705.5 Spectroscopy Cells Cultured Cultured Chemistry Vascular Endothelial Growth Factor General Medicine Computer Science Applications Endothelial stem cell Vascular endothelial growth factor A medicine.anatomical_structure High glucose Signal Transduction medicine.medical_specialty Endothelium Phospholipase A2 Inhibitors Cells Recombinant Fusion Proteins Blood–retinal barrier Arachidonic Acids Catalysis Article Inorganic Chemistry Downregulation and upregulation Retinal endothelium Vascular Internal medicine medicine Humans Physical and Theoretical Chemistry Molecular Biology Cell damage Diabetic Retinopathy Organic Chemistry Endothelial Cells Retinal Aflibercept Endothelium Vascular Glucose Phospholipases A2 Receptors Vascular Endothelial Growth Factor medicine.disease Endocrinology lcsh:Biology (General) lcsh:QD1-999 |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 21, Iss 7528, p 7528 (2020) Volume 21 Issue 20 |
| Popis: | Early blood retinal barrier (BRB) dysfunction induced by hyperglycemia was related to increased pro-inflammatory activity of phospholipase A2 (PLA2) and the upregulation of vascular endothelial growth factor A (VEGF-A). Here, we tested the role of VEGF-A in high glucose (HG)-induced damage of human retinal endothelial cells (HRECs) mediated by Ca++-dependent (cPLA2) and Ca++-independent (iPLA2) PLA2s. HRECs were treated with normal glucose (5 mM, NG) or high glucose (25 mM, HG) for 48 h with or without the VEGF-trap Aflibercept (Afl, 40 µ g/mL), the cPLA2 inhibitor arachidonoyl trifluoromethyl ketone (AACOCF3 15 µ M), the iPLA2 inhibitor bromoenol lactone (BEL 5 µ M), or VEGF-A (80 ng/mL). Both Afl and AACOCF3 prevented HG-induced damage (MTT and LDH release), impairment of angiogenic potential (tube-formation), and expression of VEGF-A mRNA. Furthermore, Afl counteracted HG-induced increase of phospho-ERK and phospho-cPLA2 (immunoblot). VEGF-A in HG-medium increased glucose toxicity, through upregulation of phospho-ERK, phospho-cPLA2, and iPLA2 (about 55%, 45%, and 50%, respectively) immunocytochemistry confirmed the activation of these proteins. cPLA2 knockdown by siRNA entirely prevented cell damage induced by HG or by HG plus VEGF-A, while iPLA2 knockdown produced a milder protective effect. These data indicate that VEGF-A mediates the early glucose-induced damage in retinal endothelium through the involvement of ERK1/2/PLA2 axis activation. |
| Databáze: | OpenAIRE |
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