Exogenous NG-hydroxy-l-arginine causes nitrite production in vascular smooth muscle cells in the absence of nitric oxide synthase activity

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Abstract

Nitric oxide (NO) production from exogenous NG-hydroxy-l-arginine (OH-l-Arg) was investigated in rat aortic smooth muscle cells in culture by measuring nitrite accumulation in the culture medium. As well, the interaction between OH-l-Arg and l-arginine uptake via the y+ cationic amino acid transporter was studied. In cells without NO-synthase activity, OH-l-Arg (1-1000 μM) induced a dose-dependent nitrite production with a half-maximal effective concentration (EC50) of 18.0 ± 1.5 μM (n = 4-7). This nitrite accumulation was not inhibited by the NO-synthase inhibitor NG-nitro-l-arginine methyl ester, l-NAME (300 μM). In contrast, it was abolished by miconazole (100 μM), an inhibitor of cytochrome P450. Incubation of vascular smooth muscle cells with LPS (10 μg ml) induced an l-name inhibited nitrite accumulation, but did not enhance the OH-l-Arg induced nitrite production. OH-l-Arg and other cationic amino acids, L-lysine and l-ornithine, competitively inhibited [3H]-l-arginine uptake m rat aortic smooth muscle cells, with inhibition constants of 195 ± 23 μM(n = 12), 260 ± 40 μM(n= 5) and 330 ± 10 μM(n = 5), respectively. These results show that OH-l-Arg is recognized by the cationic l-amino acid carrier present in vascular smooth muscle cells and can be oxidized to NO and nitrite in these cells in the absence of NO-synthase, probably by cytochrome P450 or by a reaction involving a cytochrome P450 byproduct. © 1994.

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Schott, C. A., Bogen, C. M., Vetrovsky, P., Berton, C. C., & Stoclet, J. C. (1994). Exogenous NG-hydroxy-l-arginine causes nitrite production in vascular smooth muscle cells in the absence of nitric oxide synthase activity. FEBS Letters, 341(2–3), 203–207. https://doi.org/10.1016/0014-5793(94)80457-5

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