A High-Salt Diet Dissociates NO Synthase-3 Expression and NO Production by the Thick Ascending Limb

M. Herrera, G. Silva, J. L. Garvin
2005 Hypertension  
NO produced by endothelial NO synthase (NOS3) decreases sodium transport by the thick ascending limb (THAL). We found previously that 7 days of high salt (HS) increased THAL-NOS3 expression but not NO production. NOS3 phosphorylation regulates enzyme activity. We hypothesized that HS acutely increases NOS3 expression and NO production, and, over time, changes in NOS3 phosphorylation dissociate NO production from expression. NOS3 expression increased by 71Ϯ13%, 127Ϯ24%, and 69Ϯ16% at days 1, 3,
more » ... Ϯ16% at days 1, 3, and 7 of HS, respectively. At days 14 and 28, expression was back to normal salt. After 1 day of HS, NO production in response to 250 mol/L L-arginine was elevated by 146% and, by day 3, returned to normal salt. Similar increases were found in response to endothelin-1. Inhibitors of NOS1/2 did not blunt the salt-induced increase in NO. Phosphorylation at Thr 495 , an inhibitory site, decreased by 39Ϯ8% at day 1 of HS and then increased by 116Ϯ18% at day 3. Phosphorylation at Ser 633 and Ser 1177 (stimulatory sites) decreased by Ϸ25% at day 1 and remained depressed at day 3. Superoxide production increased by 71% at day 1, decreased by 57% at day 3, and decreased by 55% at day 7. The NOS inhibitor L-N G -nitroarginine methyl ester did not alter superoxide levels at any time point. The addition of reduced nicotinamide-adenine dinucleotide phosphate and tetrahydrobiopterin had no effect on NO release after 3 days of HS. We conclude the following: (1) HS transiently increases NO production and NOS3 expression; (2) NOS3 expression and NO production are dissociated by HS; and (3) changes in phosphorylation explain how THAL NOS3 activity and expression are dissociated by HS. (Hypertension. 2006;47:95-101.)
doi:10.1161/01.hyp.0000196274.78603.85 pmid:16344378 fatcat:vze6zgih6fhfvifxlishu77acm