Neuroprotection of Rodent Retinal Ganglion Cells using Hybrid Molecule SA-10

Date

2023

Authors

Pham, Jennifer H.
Kodati, Bindu
Johnson, Gretchen A.
Acharya, Suchismita
Stankowska, Dorota L.

ORCID

0000-0003-0965-3625 (Pham, Jennifer H.)

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Abstract

Purpose: Oxidative stress is the imbalance between the activity of antioxidants and free radical production, which has been shown to be associated with glaucomatous retinal ganglion cell (RGC) degeneration. In this study, we aimed to promote RGC survival by treatment with SA-10, a second-generation hybrid molecule with nitric oxide donating and sulfone reactive oxygen species (ROS) scavenging moieties in vitro and ex vivo following oxidative stress-induced injury.

Methods: Endothelin-3, a vasoactive peptide, was used to induce oxidative stress in vitro in rat primary RGCs (n=3 biological replicates) and ex vivo in C57BL/6J mice retinal explants (n=8-9 explants/group). Primary RGCs were isolated from Sprague Dawley rat pups (post-natal days 4-7) and cultured for seven days with neurotrophic factors to allow for neurite outgrowth. The RGCs and retinal explants were pretreated with vehicle (DPBS) or SA-10 [10 µM] for 30 minutes, following which ET-3 treatment [100 nM or 400 nM] was carried out for 1 hour. CellROX™ Green was then used to stain for ROS produced by the cells, and the integrated density was analyzed. Analysis of Variance (ANOVA) or nonparametric Kruskal-Wallis was performed for all experiments.

Results: In primary RGCs, ET-3-mediated ROS production decreased by 25.9% (p<0.01) following SA-10 treatment compared to the vehicle. In mice, retinal explants, 400 nM ET-3 induced a 24.4% increase in ROS production compared to the vehicle [0 nM ET-3]. With the SA-10 treatment, the ROS production was decreased by 14.74% (p<0.001) in the ET-3 and SA-10 treated group compared to the ET-3-only treated group.

Conclusion: SA-10 effectively protects rodent RGCs in vitro and ex vivo from ET-3-mediated oxidative stress.

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