A high-salt diet stimulates thick ascending limb eNOS expression by raising medullary osmolality and increasing release of endothelin-1

A high-salt diet increases renal endothelin (ET) production and thick ascending limb (THAL) endothelial nitric oxide synthase ( eNOS) expression. ET stimulates THAL eNOS expression via ETB receptors. The tonicity of the renal medulla is highly variable, and hyperosmolality stimulates ET-1 synthesis by endothelial cells. We hypothesized that a high-salt diet raises medullary osmolality, increases ET release by the THAL, and thus enhances eNOS expression. Seven days of high salt (1% NaCl in drinking water) increased eNOS expression in THALs by 125 +/- 31%. High salt increased outer medullary osmolality from 362 +/- 13 to 423 +/- 6 mosmol/kgH(2)O ( P < 0.05). Bosentan, a dual-ET receptor antagonist, blocked the increase in THAL eNOS expression caused by high salt (2.66 +/- 0.44 absorbance units with bosentan vs. 5.15 +/- 0.67 for vehicle; P < 0.05). Conscious systolic blood pressure did not differ between the two groups. In primary cultures of medullary THALs, raising osmolality from 300 to 350 and 400 mosmol/kgH(2)O using NaCl increased eNOS expression by 39 +/- 11% (P < 0.05) and 71 +/- 16%, respectively (P < 0.05). In primary cultures of THALs, raising osmolality from 300 to 400 mosmol/kgH(2)O for 1 h increased ET-1 release from 62 +/- 7 to 113 +/- 2 pg/mg protein (P < 0.05). BQ-788, an ETB receptor antagonist (1 mu M), blocked the stimulatory effect of 400 mosmol/kgH(2)O on eNOS expression (70 +/- 13% vs. - 5 +/- 10%; paired difference, 74 +/- 15%; P < 0.05). BQ-788 alone had no significant effect. We concluded that high salt stimulates THAL eNOS expression by increasing outer medullary osmolality, ET-1 release by the THAL and ETB receptor activation. This may be an important regulatory mechanism of THAL NaCl absorption when dietary salt intake is increased.