J Korean Ophthalmol Soc.  2013 Sep;54(9):1429-1434.

A Study of the Pathway of Nitric Oxide Production by Nitroglycerin in Trabecular Meshwork Cells

Affiliations
  • 1Department of Ophthalmology, Catholic University of Daegu School of Medicine, Daegu, Korea. jwkim@cu.ac.kr

Abstract

PURPOSE
To investigate the effects of nitroglycerin on the production of nitric oxide and its related pathway in cultured human trabecular meshwork cells (HTMC).
METHODS
Primarily cultured HTMC were exposed to 10 nM nitroglycerin using 1% serum-containing media for 30 minutes. The production of nitric oxide was assessed with the Griess assay and expressions of eNOS mRNA was assessed with RT-PCR. Additionally, the cells were exposed to wortmanin and Akt1/2 kinase inhibitor to investigate the mechanism related to the production of nitric oxide.
RESULTS
Nitroglycerin increased the production of nitric oxide (p < 0.05) accompanied with increased expression of eNOS mRNA. The increased production of nitric oxide and eNOS mRNA was inhibited by wortmanin and Akt1/2 kinase inhibitor.
CONCLUSIONS
Low-dose nitroglycerin increased the production of nitric oxide accompanied by increased eNOS activity. Nitroglycerin drives eNOS activation via the phosphatidylinositol 3-kinase/protein kinase B pathway.

Keyword

eNOS; Nitroglycerin; Nitric oxide; Trabecular meshwork cells

MeSH Terms

Humans
Nitric Oxide
Nitroglycerin
Phosphatidylinositols
Phosphotransferases
RNA, Messenger
Trabecular Meshwork
Nitric Oxide
Nitroglycerin
Phosphatidylinositols
Phosphotransferases
RNA, Messenger

Figure

  • Figure 1. Effects of Nitroglycerin (GTN) and vascular endo-thelial growth factor (VEGF) on the production of nitric oxide in trabecular meshwork cells. Both GTN and VEGF increased production of nitric oxide significantly (* p < 0.05).

  • Figure 2. Effects of wortmanin and Akt1/2 kinase inhibitor on the nitroglycerin (GTN)-induced production of nitric oxide in trabecular meshwork cells. Akt1/2 kinase inhibitor and wort-manin inhibits GTN-induced production of nitric oxide (* p < 0.05).

  • Figure 3. Effects of nitroglycerin (GTN) on the activity of eNOS in trabecular meshwork cells. GTN and vascular growth factor (VEGF) increased eNOS expression, which were abolished by Akt1/2 kinase inhibitor and wortmanin compared to the non-exposed control.


Reference

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