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
Abstract
:1. Introduction
2. Results
2.1. Effects of Aflibercept and PLA2 Inhibitors on Retinal Endothelial Cell Damage Induced by HG
2.2. Effect of Aflibercept and the cPLA2 Inhibitor AACOCF3 on Tube-Like Structure Formation in HRECs Impaired by HG
2.3. Effects of Aflibercept and PLA2 Inhibitors on VEGF-A/PLA2 Axis Activation in HRECs Stimulated by HG
2.4. Effects of Exogenous VEGF-A in HG-Stimulated HREC
2.5. Activation of ERK1/2/cPLA2 Pathway and iPLA Expression by VEGF-A in Presence of HG in HREC
2.6. Selective PLA2 Silencing Reduces the Detrimental Effects of HG in HRECs
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Cultures and Treatments
4.3. Cell Viability Assay
4.4. Lactate Dehydrogenase (LDH) Release
4.5. Immunocytochemical Analysis
4.6. Tube Formation Assay
4.7. Western Blot Analysis
4.8. Transfection of siRNA
4.9. Real-Time Reverse Transcriptase-Polymerase Chain Reaction (RT-qPCR)
4.10. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Giurdanella, G.; Lupo, G.; Gennuso, F.; Conti, F.; Furno, D.L.; Mannino, G.; Anfuso, C.D.; Drago, F.; Salomone, S.; Bucolo, C. 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. Int. J. Mol. Sci. 2020, 21, 7528. https://doi.org/10.3390/ijms21207528
Giurdanella G, Lupo G, Gennuso F, Conti F, Furno DL, Mannino G, Anfuso CD, Drago F, Salomone S, Bucolo C. 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. International Journal of Molecular Sciences. 2020; 21(20):7528. https://doi.org/10.3390/ijms21207528
Chicago/Turabian StyleGiurdanella, Giovanni, Gabriella Lupo, Florinda Gennuso, Federica Conti, Debora Lo Furno, Giuliana Mannino, Carmelina Daniela Anfuso, Filippo Drago, Salvatore Salomone, and Claudio Bucolo. 2020. "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" International Journal of Molecular Sciences 21, no. 20: 7528. https://doi.org/10.3390/ijms21207528