Stefano De De Cillà1, Serena Farruggio2, Stela Vujosevic3, Giulia Raina2, Davide Filippini2, Valentina Gatti1,2, Nausicaa Clemente4, David Mary2, Diego Vezzola1, Giamberto Casini5, Luca Rossetti6, Elena Grossini2. 1. Oculistic Unit, Dept. Health Sciences, University East Piedmont, Azienda Ospedaliera Universitaria Maggiore della Carità, Novara, Italy. 2. Lab. Physiology/Experimental Surgery, Dept. Translational Medicine, University East Piedmont, Novara, Italy. 3. Eye Clinic, Azienda Ospedaliera Universitaria Maggiore della Carità, Novara, Italy. 4. Lab. Immunology, IRCAD, Dept. Health Sciences, University East Piedmont, Novara, Italy. 5. Dept. Surgical, Medical, Molecular and Critical Area Pathology, University of Pisa, Pisa, Italy. 6. Eye Clinic, San Paolo Hospital, University of Milan, Milan, Italy.
Abstract
BACKGROUND/AIMS: the anti-vascular endothelial growth factors (VEGF), Aflibercept and Ranibizumab, are used for the treatment of macular degeneration. Here we examined the involvement of nitric oxide (NO), mitochondria function and of apoptosis/autophagy in their antioxidant effects in human retinal pigment epithelium cells (RPE). METHODS: RPE were exposed to Ranibizumab/Aflibercept in the absence or presence of NO synthase (NOS) inhibitor and of autophagy activator/blocker, rapamicyn/3-methyladenine. Specific kits were used for cell viability, NO and reactive oxygen species detection and mitochondrial membrane potential measurement, whereas Western Blot was performed for apoptosis/ autophagy markers and other kinases detection. RESULTS: In RPE cultured in physiological conditions, Aflibercept/Ranibizumab increased NO release in a dose and time-dependent way. Opposite results were obtained in RPE pretreated with hydrogen peroxide. Moreover, both the anti-VEGF agents were able to prevent the fall of cell viability and of mitochondrial membrane potential. Those effects were reduced by the NOS inhibitor and 3-methyladenine and were potentiated by rapamycin. Finally, Aflibercept and Ranibizumab counteracted the changes of apoptosis/autophagy markers, NOS, Phosphatidylinositol-3-Kinase/Protein Kinase B and Extracellular signal-regulated kinases 1/2 caused by peroxidation. CONCLUSION: Aflibercept and Ranibizumab protect RPE against peroxidation through the modulation of NO release, apoptosis and autophagy.
BACKGROUND/AIMS: the anti-vascular endothelial growth factors (VEGF), Aflibercept and Ranibizumab, are used for the treatment of macular degeneration. Here we examined the involvement of nitric oxide (NO), mitochondria function and of apoptosis/autophagy in their antioxidant effects in human retinal pigment epithelium cells (RPE). METHODS: RPE were exposed to Ranibizumab/Aflibercept in the absence or presence of NO synthase (NOS) inhibitor and of autophagy activator/blocker, rapamicyn/3-methyladenine. Specific kits were used for cell viability, NO and reactive oxygen species detection and mitochondrial membrane potential measurement, whereas Western Blot was performed for apoptosis/ autophagy markers and other kinases detection. RESULTS: In RPE cultured in physiological conditions, Aflibercept/Ranibizumab increased NO release in a dose and time-dependent way. Opposite results were obtained in RPE pretreated with hydrogen peroxide. Moreover, both the anti-VEGF agents were able to prevent the fall of cell viability and of mitochondrial membrane potential. Those effects were reduced by the NOS inhibitor and 3-methyladenine and were potentiated by rapamycin. Finally, Aflibercept and Ranibizumab counteracted the changes of apoptosis/autophagy markers, NOS, Phosphatidylinositol-3-Kinase/Protein Kinase B and Extracellular signal-regulated kinases 1/2 caused by peroxidation. CONCLUSION: Aflibercept and Ranibizumab protect RPE against peroxidation through the modulation of NO release, apoptosis and autophagy.
Authors: Salvador Mérida; Vincent M Villar; Amparo Navea; Carmen Desco; María Sancho-Tello; Cristina Peris; Francisco Bosch-Morell Journal: Front Physiol Date: 2020-05-14 Impact factor: 4.566
Authors: Elena Grossini; Divya Praveen Garhwal; Giuseppe Calamita; Raffaele Romito; Cristina Rigamonti; Rosalba Minisini; Carlo Smirne; Daniela Surico; Mattia Bellan; Mario Pirisi Journal: Front Med (Lausanne) Date: 2021-07-02