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Scientific Reports

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Ripa-56 protects retinal ganglion cells in glutamate-induced retinal excitotoxic model of glaucoma

Published:15 February 2024 DOI: 10.1038/s41598-024-54075-z PMID: 38360971
Lemeng Feng,?Shirui Dai,?Cheng Zhang,?Wulong Zhang,?Weiming Zhu,?Chao Wang,?Ye He,?Weitao Song

Abstract

Glaucoma is a prevalent cause of blindness globally, characterized by the progressive degeneration of retinal ganglion cells (RGCs). Among various factors, glutamate excitotoxicity stands out as a significant contributor of RGCs loss in glaucoma. Our study focused on Ripa-56 and its protective effect against NMDA-induced retinal damage in mice, aiming to delve into the potential underlying mechanism. The R28 cells were categorized into four groups: glutamate (Glu), Glu?+?Ripa-56, Ripa-56 and Control group. After 24?h of treatment, cell death was assessed by PI / Hoechst staining. Mitochondrial membrane potential changes, apoptosis and reactive oxygen species (ROS) production were analyzed using flow cytometry. The alterations in the expression of RIP-1, p-MLKL, Bcl-2, BAX, Caspase-3, Gpx4 and SLC7A11 were examined using western blot analysis. C57BL/6j mice were randomly divided into NMDA, NMDA?+?Ripa-56, Ripa-56 and control groups. Histological changes in the retina were evaluated using hematoxylin and eosin (H&E) staining. RGCs survival and the protein expression changes of RIP-1, Caspase-3, Bcl-2, Gpx4 and SLC7A11 were observed using immunofluorescence. Ripa-56 exhibited a significant reduction in the levels of RIP-1, p-MLKL, Caspase-3, and BAX induced by glutamate, while promoting the expression of Bcl-2, Gpx-4, and SLC7A1 in the Ripa-56-treated group. In our study, using an NMDA-induced normal tension glaucoma mice model, we employed immunofluorescence and H&E staining to observe that Ripa-56 treatment effectively ameliorated retinal ganglion cell loss, mitigating the decrease in retinal ganglion cell layer and bipolar cell layer thickness caused by NMDA. In this study, we have observed that Ripa-56 possesses remarkable anti- necroptotic, anti-apoptotic and anti-ferroptosis properties. It demonstrates the ability to combat not only glutamate-induced excitotoxicity in R28 cells, but also NMDA-induced retinal excitotoxicity in mice. Therefore, Ripa-56 could be used as a potential retinal protective agent.

Substances (4)

Materials
Procduct Name CAS Molecular Formula Supplier Price
RIPA-56 1956370-21-0 C13H19NO2 77 suppliers Inquiry
RIPA-56 1956370-21-0 C13H19NO2 77 suppliers Inquiry
RIPA-56 1956370-21-0 C13H19NO2 77 suppliers Inquiry
RIPA-56 1956370-21-0 C13H19NO2 77 suppliers Inquiry

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