Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy
Abstract
:1. Introduction
2. Materials and Methods
2.1. In Vitro Studies
2.1.1. MIO-M1 Cell Culture
2.1.2. Cell Viability/Proliferation
2.1.3. Pharmacological Treatments
2.1.4. Preparation of Conditioned Medium
2.1.5. Quantitative Real-Time PCR
2.1.6. Enzyme-Linked Immunosorbent Assay (ELISA)
2.1.7. Immunofluorescence
2.2. Ex Vivo Studies
2.2.1. Preparation of Retinal Explants
2.2.2. Pharmacological Treatments
2.2.3. Preparation of Conditioned Media
2.2.4. Quantitative Real-Time PCR
2.2.5. ELISA
2.2.6. Immunofluorescence
2.3. Statistics
3. Results
3.1. OS Induces VEGFR2-Dependent VEGF Expression Both in MIO-M1 Cells and in Retinal Explants
3.2. Exogenous VEGF Induces VEGFR2-Dependent VEGF Expression Both in MIO-M1 Cells and in Retinal Explants
3.3. Conditioned Medium Induces VEGF Expression in MIO-M1 Cells
3.4. Conditioned Medium Does Not Induce VEGF Expression in Retinal Explants
3.5. OS Triggers Nrf2 Nuclear Translocation, while OS-Induced VEGF Expression Is Inhibited by Nrf2 Blockade
3.6. Exo-VEGF Has No Effect on Nrf2 Nuclear Translocation, while Exo-VEGF-Induced VEGF Expression Is Not Affected by Nrf2 Blockade
3.7. Blockade of HIF-1 Inhibits VEGF mRNA Expression in All Conditions
4. Discussion
4.1. The Role of OS
4.2. The Involvement of VEGFR2
4.3. The Involvement of Müller Cells
4.4. Intracellular Pathways
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Rossino, M.G.; Lulli, M.; Amato, R.; Cammalleri, M.; Dal Monte, M.; Casini, G. Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy. Cells 2020, 9, 1452. https://doi.org/10.3390/cells9061452
Rossino MG, Lulli M, Amato R, Cammalleri M, Dal Monte M, Casini G. Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy. Cells. 2020; 9(6):1452. https://doi.org/10.3390/cells9061452
Chicago/Turabian StyleRossino, Maria Grazia, Matteo Lulli, Rosario Amato, Maurizio Cammalleri, Massimo Dal Monte, and Giovanni Casini. 2020. "Oxidative Stress Induces a VEGF Autocrine Loop in the Retina: Relevance for Diabetic Retinopathy" Cells 9, no. 6: 1452. https://doi.org/10.3390/cells9061452