The effect of inhaled nitric oxide and oxygen on the hydroxylation of salicylate in rat lungs

Inhaled nitric oxide (iNO) is used as a selective pulmonary vasodilator, and often under conditions when a high fraction of inspired oxygen is indicated. However, little is known about the potential toxicity of iNO therapy with or without concomitant oxygen therapy. NO can combine with superoxide (O2-) to form peroxynitrite (ONOO-), which can in turn decompose to form hydroxyl radical (OH.). Both OH. and ONOO- are involved in various forms of lung injury. To begin evaluation of the effect of iNO under either normoxic or hyperoxic conditions on OH. and/or ONOO- formation, rats were exposed for 58 h to either 21% O2, 21% O2 + 10 parts per million (ppm) NO, 21% O2 + 100 ppm NO, 50% O2, 90% O2, 90% O2 + 10 ppm NO, or 90% O2 + 100 ppm NO. We used a salicylate hydroxylation assay to detect the effects of these exposures on lung OH. and/or ONOO- formation measured as the appearance of 2,3-dihydroxybenzoic acid (2,3-DHBA). Exposure to 90% O2 and 90% O2 + 100 ppm NO resulted in significantly (p < 0.05) greater lung wet weight (1.99 +/- 0.14 g and 3.14 +/- 0.30 g, respectively) compared with 21% O2 (1.23 +/- 0.01 g). Exposure to 21% O2 + 100 ppm NO led to 2.5 times the control (21% O2 alone) 2,3 DHBA formation (p < 0.05) and exposure to 90% O2 led to 2.4 times the control 2,3-DHBA formation (p < 0.05). However, with exposure to both 90% O2 and 100 ppm NO, the 2,3-DHBA formation was no greater than the control condition (21% O2). Thus, these results indicate that, individually, both the hyperoxia and the 100 ppm NO led to greater salicylate hydroxylation, but that the combination of hyperoxia and 100 ppm NO led to less salicylate hydroxylation than either did individually. The production of OH. and/or ONOO- in the lung during iNO therapy may depend on the ratio of NO to O2.