섬유주세포에서 니트로글리세린에 의한 일산화질소의 생성경로에 관한 연구
A Study of the Pathway of Nitric Oxide Production by Nitroglycerin in Trabecular Meshwork Cells
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목적: 섬유주세포에서 니트로글리세린이 일산화질소의 생성에 미치는 영향과 기전을 알아 보고자 하였다. 대상과 방법: 섬유주세포를 일차배양한 후 1% 혈청이 포함된 배지를 이용하여 10 nM의 니트로글리세린에 30분간 노출시킨 후 Griess assay를 이용하여 일산화질소의 생성량을 측정하였으며, RT-PCR을 이용하여 eNOS mRNA의 발현을 조사하였다. 이때 니트로글리세린의 작용기전을 알아보기 위하여 wortmanin과 Akt1/2 kinase 저해제에도 노출시켰다. 결과: 니트로글리세린은 섬유주세포에서 일산화질소의 생성을 유의하게 증가시켰으며(p<0.05), 이러한 생성증가는 wortmanin과Akt1/2 kinase 저해제에 의해 유의하게 억제되었다. 이와 함께 니트로글리세린은 eNOS mRNA의 발현을 증가시켰으며 wortmanin과Akt1/2 kinase 저해제는 eNOS mRNA의 발현을 감소시켰다. 결론: 섬유주세포에서 저농도의 니트로글리세린은 일산화질소의 생성을 증가시켰으며 이러한 증가의 기전으로 PI3K/Akt 경로가 작용할 것이라 생각한다.
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.
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