Down Syndrome Critical Region 1 Reduces Oxidative Stress-Induced Retinal Ganglion Cells Apoptosis via CREB-Bcl-2 Pathway

Invest Ophthalmol Vis Sci. 2020 Oct 1;61(12):23. doi: 10.1167/iovs.61.12.23.

Abstract

Purpose: Irreversible retina ganglion cell (RGC) loss is a key process during glaucoma progression. Down syndrome critical region 1 (DSCR1) has been shown to have protective effects against neuronal death. In this study, we aimed to investigate the neuroprotective mechanisms of DSCR1 on RGCs.

Methods: DBA/2J mice and optic nerve crush (ONC) rat model were used for vivo assays. Oxidative stress model of primary RGCs was carried out with in vitro transduction. DSCR1 protein localization was assessed by immunofluorescence. Differential protein expression was validated by Western blot, and gene expression was detected by real-time PCR. TUNEL was used to identify cell apoptosis, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide was used to analyze cell viability.

Results: Significant upregulation of DSCR1 was observed in DBA/2J mice, ONC rat model, and RGCs treated with H2O2, reaching peaks at the age of 6 months in DBA/2J mice, 5 days after ONC in rats, and 24 hours after H2O2 treatment in RGCs, respectively. DSCR1 was shown to be expressed in the ganglion cell layer. In vitro, overexpressed DSCR1 significantly promoted phosphorylation of cyclic AMP response element binding protein (CREB), B-cell lymphoma 2 (Bcl-2) expression, and RGC survival rate while reducing cleaved caspase 3 expression in H2O2-treated RGCs. On the other hand, the opposite effects were shown after knockdown of DSCR1. In addition, silencing of CREB inhibited expression of DSCR1.

Conclusions: Our results suggested that DSCR1 might protect the RGCs against oxidative stress via the CREB-Bcl-2 pathway, which may provide a theoretical basis for future treatments of glaucoma.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Blotting, Western
  • Calcium-Binding Proteins / genetics*
  • Cyclic AMP Response Element-Binding Protein / metabolism*
  • Disease Models, Animal
  • Fluorescent Antibody Technique, Indirect
  • Gene Expression Regulation / physiology*
  • Hydrogen Peroxide / toxicity
  • In Situ Nick-End Labeling
  • Lentivirus / genetics
  • Mice
  • Mice, Inbred C57BL
  • Mice, Inbred DBA
  • Muscle Proteins / genetics*
  • Nerve Crush
  • Oxidative Stress*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism*
  • Rats
  • Real-Time Polymerase Chain Reaction
  • Retina / metabolism
  • Retina / pathology
  • Retinal Ganglion Cells / metabolism*
  • Retinal Ganglion Cells / pathology
  • Transfection

Substances

  • Calcium-Binding Proteins
  • Creb1 protein, mouse
  • Cyclic AMP Response Element-Binding Protein
  • DSCR1 protein, mouse
  • Muscle Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Bcl2 protein, mouse
  • Hydrogen Peroxide