Sulphur Antioxidants Inhibit Oxidative Stress Induced Retinal Ganglion Cell Death by Scavenging Reactive Oxygen Species but Influence Nuclear Factor (Erythroid-Derived 2)-like 2 Signalling Pathway Differently

Biol Pharm Bull. 2013;36(7):1095-110. doi: 10.1248/bpb.b13-00023.

Abstract

This study aimed to show if two different sulphur containing drugs sulbutiamine and acetylcysteine (NAC) could attenuate the effects of two different insults being serum deprivation and glutamate/buthionine sulfoximine (GB)-induced death to transformed retinal ganglion cell line (RGC-5) in culture. Cells were exposed to either 5 mM of GB for 24 h or serum deprivation for 48 h with inclusion of either NAC or sulbutiamine. Cell viability, microscopic evidence for apoptosis, caspase 3 activity, reactive oxygen species (ROS), glutathione (GSH), catalase and gluthathione-S-transferase (GST) were determined. The effects of NAC and sulbutiamine on the oxidative stress related transcription factor nuclear factor (erythroid-derived 2)-like 2 (Nrf-2) levels and its dependent phase II enzyme haemeoxygenase-1 (HO-1) were carried out using Western blot and quantitative-polymerase chain reaction (PCR). NAC and sulbutiamine dose-dependently attenuated serum deprivation-induced cell death. However NAC but not sulbutiamine attenuated GB-induced cell death. NAC and sulbutiamine both independently stimulated the GSH and GST production but scavenged different types of ROS with different efficacy. Moreover only sulbutiamine stimulated catalase and significantly increased Nrf-2 and HO-1 levels. In addition, the pan caspase inhibitor, benzoylcarbonyl-Val-Ala-Asp-fluoromethyl ketone (z-VAD-fmk) attenuated the negative effect of serum deprivation while the necroptosis inhibitor (necrostatin-1) counteracted solely an insult of GB. The neuroprotective actions of NAC and sulbutiamine in GB or serum-deprivation insult are therefore different.

MeSH terms

  • Acetylcysteine / pharmacology*
  • Animals
  • Apoptosis / drug effects
  • Blotting, Western
  • Cell Culture Techniques
  • Cell Line, Transformed
  • Cell Survival / drug effects
  • Free Radical Scavengers / pharmacology*
  • Heme Oxygenase (Decyclizing) / metabolism
  • NF-E2-Related Factor 2 / metabolism*
  • Oxidative Stress / drug effects*
  • Rats
  • Reactive Oxygen Species / metabolism*
  • Real-Time Polymerase Chain Reaction
  • Retinal Ganglion Cells / drug effects*
  • Retinal Ganglion Cells / metabolism
  • Signal Transduction / drug effects*
  • Thiamine / analogs & derivatives*
  • Thiamine / pharmacology

Substances

  • Free Radical Scavengers
  • NF-E2-Related Factor 2
  • Reactive Oxygen Species
  • sulbutiamine
  • Heme Oxygenase (Decyclizing)
  • Hmox1 protein, rat
  • Acetylcysteine
  • Thiamine