title: Acute and chronic regulation of thick ascending limb endothelial nitric oxide synthase by statins.
abstract: Statins, 3-hydroxy-3-methylglutaryl CoA reductase inhibitors, acutely increase endothelial nitric oxide synthase (eNOS) activity and chronically increase eNOS expression in endothelial cells. NO decreases transport in thick ascending limbs (TAL). We hypothesized that statins inhibit TAL transport by acutely activating eNOS, thereby increasing NO production and chronically enhancing eNOS expression. Oxygen consumption (QO(2)) by TAL suspensions from Sprague-Dawley rats was used as a measure of active NaCl reabsorption. Na/K ATPase activity was assessed by measuring ATP hydrolysis in the presence and absence of ouabain. eNOS expression was measured by Western blot. A total of 50 micro M pravastatin decreased QO(2) by 18.6 +/- 3.4% (P < 0.01). In the presence of 500 micro M furosemide and 200 micro M amiloride, transport blockers, QO(2) remained the same after pravastatin was added. Na/K ATPase activity was not different from controls and TAL treated with 50 micro M pravastatin (0.33 +/- 0.07 versus 0.29 +/- 0.04 nmol P(i)/ micro g protein/min, where P(i) is inorganic phosphate). Nystatin stimulated QO(2) to 178 +/- 13.7 in pravastatin-treated TAL and 195 +/- 11.5 in furosemide-treated TAL. The inhibitory effect of pravastatin on QO(2) was blocked by L-nitroarginine methyl ester, an NOS inhibitor. In addition, pravastatin increased NO production as measured by the fluorescent dye DAF-2A. Pravastatin at a dose of 10 mg/kg per d had no effect on eNOS protein at 1 d (24.1 +/- 2.7 versus 25.5 +/- 1.1 arbitrary units [AU]) or 7 d (24.1 +/- 2.7 versus 20.9 +/- 1.3 AU). Similarly, at 1 d, 50 mg/kg per d had no effect on expression (24.1 +/- 2.7 versus 21.2 +/- 3.6 AU). At 7 d, this dose decreased eNOS protein from 24.1 +/- 2.7 to 11.8 +/- 4.4 AU. It is concluded that pravastatin acutely decreases NaCl entry into the TAL by releasing NO. Pravastatin does not chronically increase eNOS expression in TAL.
authors.affiliation: Henry Ford Health System, Detroit, Michigan.
authors.forename: Marisela
authors.initials: M
authors.lastname: Varela
authors.forename: Jeffrey L
authors.initials: JL
authors.lastname: Garvin
mesh_terms: D000818:Animals; D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors / Q000494:pharmacology*; D008138:Loop of Henle / Q000187:drug effects / Q000201:enzymology*; D009569:Nitric Oxide / Q000096:biosynthesis; D019001:Nitric Oxide Synthase / Q000502:physiology*; D010101:Oxygen Consumption / Q000187:drug effects; D017035:Pravastatin / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D019161:Hydroxymethylglutaryl-CoA Reductase Inhibitors; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D017035:Pravastatin