Immunohystochemical and gene expression analysis of the effect of bevacizumab upon lister rat retinal explants and on a human Müller glial cell line

Abstract

Retinal diseases that are associated to neovascularization, such as age-related macular degeneration, diabetic retinopathy and retinopathy of prematurity are important causes of blindness throughout the world. In the last years, intravitreal injection of anti-angiogenic drugs, such as bevacizumab, has been of great value both in the adulthood as well as in newborn infants. However, experimental in vitro and in vivo studies have shown that bevacizumab has also the capacity of causing adverse ocular effects on cellular processes interfering in proliferative and differentiation retinal processes. Based on the current knowledge on pathogenesis of retinal diseases, the current study has the purpose of investigating bevacizumab effect on cellular and molecular mechanisms of the retina, particularly the differentiation mechanisms, growth factors and retinal proteoglycans that support such events. In the study, biochemical and molecular biology techniques will be employed to provide quantitative and gene expression analysis. Immunocytochemistry assays and LUMINEX immunoassays will be employed. mRNA expression will be analyzed using reverse transcriptase reaction, in which amplification of selected sequences will be performed using real-time polymerase chain reaction. Findings in this study will hopefully bring important insights for future studies regarding synthesis of markers and the effect of anti-angiogenic drugs in the retina as well as for the knowledge about mechanisms of action that will be important for designing new treatment options. (AU)