J Korean Ophthalmol Soc.  2009 Oct;50(10):1569-1575. 10.3341/jkos.2009.50.10.1569.

Effect of Methylglyoxal on the Oxidative Stress in Trabecular Meshwork Cells

• Lee SH
• Kim SH
• Kim JW

• KMID: 2212777
• DOI: http://doi.org/10.3341/jkos.2009.50.10.1569

Abstract

PURPOSE
To investigate the effect of methylglyoxal (MG), intermediate metabolite of advanced glycation end products(AGE), on the induction of oxidative stress in human trabecular meshwork cells (HTMC).
METHODS
Primarily cultured HTMC were exposed to at concentrations of 0, 30, 100, and 300 micrometer of MG for 18 hours, with or without co-exposure to N-acetyl-cysteine. Cellular survival and apoptosis were assessed by MTT assay and flow cytometry using annexin-PI double staining. Production of nitric oxide (NO), superoxide, and reactive oxygen species (ROS) was assessed by Griess assay, cytochrome c assay, and dichlorofluorescein diacetate assay, respectively.
RESULTS
MG did not affect cellular survival at concentrations under 100 micrometer, but induced apoptosis of HTMC at concentrations over 100 micrometer. MG decreased NO production, accompanied with increased superoxide production. In addition, MG increased ROS, which were abolished by N-acetylcysteine.
CONCLUSIONS
MG induced oxidative stress by decreasing NO production, accompanied by increasing superoxide and ROS productions in HTMC. AGE could induce trabecular meshwork dysfunction.

Keyword

Advanced glycation end products; Methylglyoxal; Oxidative stress; Trabecular meshwork cells

MeSH Terms

Acetylcysteine
Apoptosis
Cytochromes c
Flow Cytometry
Glycosylation End Products, Advanced
Humans
Nitric Oxide
Oxidative Stress
Pyruvaldehyde
Reactive Oxygen Species
Superoxides
Trabecular Meshwork
Acetylcysteine
Cytochromes c
Glycosylation End Products, Advanced
Nitric Oxide
Pyruvaldehyde
Reactive Oxygen Species
Superoxides

Figure

• 

  Figure 1. Effect of methylglyoxal (MG) on the survival of trabecular meshwork cells. MG and co-exposure of 50 μM N-acetyl-cysteine (NAC) did not affect on the cellular survival significantly except at 300 μM MG. (* p<0.05)

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  Figure 2. Effect of methylglyoxal on the induction of apoptosis in cultured trabecular meshwork cells. Methylglyoxal induced apoptosis from 100 μM. (* p<0.05)

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  Figure 3. Effect of methylglyoxal (MG) on the generation of nitric oxide in cultured trabecular meshwork cells. MG decreased nitric oxide production from 30 μM. (* p<0.05)

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  Figure 4. Effect of methylglyoxal on the generation of superoxide. Methylglyoxal increased superoxide production from 30 μM. (* p<0.05)

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  Figure 5. Effect of methylglyoxal (MG) on the generation of reactive oxygen species. MG increased reactive oxygen species, which were abolished by co-exposed N-acetyl cysteine (NAC). (*,** p<0.05)

Reference

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