Inhibition of BDNF-AS Provides Neuroprotection for Retinal Ganglion Cells against Ischemic Injury

PLoS One. 2016 Dec 9;11(12):e0164941. doi: 10.1371/journal.pone.0164941. eCollection 2016.

Abstract

Background: Brain-derived neurotrophic factor (BDNF) protects retinal ganglion cells against ischemia in ocular degenerative diseases. We aimed to determine the effect of BDNF-AS on the ischemic injury of retinal ganglion cells.

Methods: The levels of BDNF and BDNF-AS were measured in retinal ganglion cells subjected to oxygen and glucose deprivation. The lentiviral vectors were constructed to either overexpress or knock out BDNF-AS. The luciferase reporter gene assay was used to determine whether BDNF-AS could target its seed sequence on BDNF mRNA. The methyl thiazolyl tetrazolium assay was used to determine cell viability, and TUNEL staining was used for cell apoptosis.

Results: The levels of BDNF-AS were negatively correlated with BDNF in ischemic retinal ganglion cells. BDNF-AS directly targeted its complementary sequences on BDNF mRNA. BDNF-AS regulated the expression of BDNF and its related genes in retinal ganglion cells. Down-regulation of BDNF-AS increased cell viability and decreased the number of TUNEL-positive retinal ganglion cells under oxygen and glucose deprivation conditions.

Conclusion: Inhibition of BDNF-AS protected retinal ganglion cells against ischemia by increasing the levels of BDNF.

MeSH terms

  • Animals
  • Animals, Newborn
  • Apoptosis / drug effects
  • Brain-Derived Neurotrophic Factor / genetics*
  • Brain-Derived Neurotrophic Factor / metabolism
  • Cell Hypoxia
  • Cell Survival / drug effects
  • Gene Expression Regulation
  • Genes, Reporter
  • Genetic Vectors / chemistry*
  • Genetic Vectors / metabolism
  • Glucose / deficiency
  • Lentivirus / genetics
  • Lentivirus / metabolism
  • Luciferases / genetics
  • Luciferases / metabolism
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neuroprotective Agents / chemistry*
  • Neuroprotective Agents / metabolism
  • Oxygen / metabolism
  • Oxygen / pharmacology
  • Primary Cell Culture
  • RNA, Long Noncoding / antagonists & inhibitors
  • RNA, Long Noncoding / genetics*
  • RNA, Long Noncoding / metabolism
  • RNA, Messenger / genetics*
  • RNA, Messenger / metabolism
  • Retinal Ganglion Cells / cytology
  • Retinal Ganglion Cells / drug effects
  • Retinal Ganglion Cells / metabolism*

Substances

  • Brain-Derived Neurotrophic Factor
  • Neuroprotective Agents
  • RNA, Long Noncoding
  • RNA, Messenger
  • long noncoding RNA BDNF-AS, mouse
  • Luciferases
  • Glucose
  • Oxygen

Grants and funding

The authors received no specific funding for this work.