J Korean Ophthalmol Soc.  2010 Feb;51(2):276-281. 10.3341/jkos.2010.51.2.276.

Effect of Glucose on the Production of Reactive Oxygen Species in Retinal Pigment Epithelial Cells

• Lee JH
• Kim JW

• KMID: 2213152
• DOI: http://doi.org/10.3341/jkos.2010.51.2.276

Abstract

PURPOSE
To investigate the effect of high glucose (HG) on the production of reactive oxygen species (ROS) in retinal pigment epithelial (RPE) cells.
METHODS
ARPE-19 cells were exposed to low glucose (5 mM) and high glucose (HG, 25 mM) for three days. Additionally, 50 micrometer N-acetyl cysteine (NAC) and 100 micrometer L-arginine were co-exposed. Productions of nitric oxide (NO), ROS, and superoxide were determined by Griess assay, DCFH-DA assay, and modified cytochrome c assay, respectively.
RESULTS
HG increased production of NO, ROS, and superoxide. HG-induced cells increased production of superoxide and ROS but were suppressed by NAC and L-arginine (substrate for NO production) as an antioxidant.
CONCLUSIONS
HG increased ROS production in RPE cells. Thus, HG may cause cellular dysfunction and damage by inducing oxidative stress in RPE cells.

Keyword

High glucose; Oxidative stress; Reactive oxygen species; Retinal pigment epithelial cells

MeSH Terms

Arginine
Cysteine
Cytochromes c
Epithelial Cells
Fluoresceins
Glucose
Nitric Oxide
Oxidative Stress
Reactive Oxygen Species
Retinaldehyde
Superoxides
Arginine
Cysteine
Cytochromes c
Fluoresceins
Glucose
Nitric Oxide
Reactive Oxygen Species
Retinaldehyde
Superoxides

Figure

• 

  Figure 1. Effect of high glucose on the survival of ARPE-19 cells. High glucose (HG, 25 mM) increased cellular survival significantly compared with low glucose (LG, 5 mM) (* p<0.05). Co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg) did not affect the survival(p>0.05).

• 

  Figure 2. Effect of high glucose on the production of nitric oxide in ARPE-19 cells. High glucose (HG, 25 mM) increased nitric oxide production significantly compared with low glucose (LG, 5 mM) (* p<0.05). Co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg) did not affect the production of nitric oxide compared to HG alone (p>0.05).

• 

  Figure 3. Effect of high glucose on the production of superoxide in ARPE-19 cells. High glucose (HG, 25 mM) increased superoxide production significantly compared with low glucose (LG, 5 mM) (* p<0.05). HG-induced increased production of superoxide was abolished by co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg), respectively (** p<0.05).

• 

  Figure 4. Effect of high glucose on the production of reactive oxygen species (ROS) in ARPE-19 cells. High glucose (HG, 25 mM) increased ROS production significantly compared with low glucose (LG, 5 mM) (* p<0.05). HG-induced increased production of ROS was abolished by co-exposed 50 μM N-acetyl cysteine (NAC) or 100 μM L-arginine (L-Arg), respectively(** p<0.05).

Reference

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