Benzamil-mediated urine alkalization is caused by the inhibition of H + -K + -ATPases

• Published in: American journal of physiology. Renal physiology 2021-04, Vol.320 (4), p.F596-F607
• Main Authors: Ayasse, Niklas, Berg, Peder, Andersen, Jesper Frank, Svendsen, Samuel Levi, Sørensen, Mads V, Fedosova, Natalya U, Lynch, I Jeanette, Wingo, Charles S, Leipziger, Jens
• Format: Article
• Language: English
• Subjects: Acids; Acute effects; Amiloride - analogs & derivatives; Amiloride - pharmacology; Animals; Collecting duct; Dietary intake; Epithelial Sodium Channels - drug effects; Epithelial Sodium Channels - metabolism; Excretion; H(+)-K(+)-Exchanging ATPase - drug effects; H(+)-K(+)-Exchanging ATPase - metabolism; Hydrogen; Kidney Tubules, Collecting - metabolism; Kidneys; Mice; Natriuresis - drug effects; Nutrient deficiency; pH effects; Renal Elimination - drug effects; Renal Elimination - physiology; Renal function; Sodium; Sodium - metabolism; Sodium, Dietary - metabolism; Urine
• ISSN: 1931-857X; 1522-1466
• DOI: 10.1152/ajprenal.00444.2020

Epithelial Na channel (ENaC) blockers elicit acute and substantial increases of urinary pH. The underlying mechanism remains to be understood. Here, we evaluated if benzamil-induced urine alkalization is mediated by an acute reduction in H secretion via renal H -K -ATPases (HKAs). Experiments were performed in vivo on HKA double-knockout and wild-type mice. Alterations in dietary K intake were used to change renal HKA and ENaC activity. The acute effects of benzamil (0.2 µg/g body wt, sufficient to block ENaC) on urine flow rate and urinary electrolyte and acid excretion were monitored in anesthetized, bladder-catheterized animals. We observed that benzamil acutely increased urinary pH (ΔpH: 0.33 ± 0.07) and reduced NH and titratable acid excretion and that these effects were distinctly enhanced in animals fed a low-K diet (ΔpH: 0.74 ± 0.12), a condition when ENaC activity is low. In contrast, benzamil did not affect urine acid excretion in animals kept on a high-K diet (i.e., during high ENaC activity). Thus, urine alkalization appeared completely uncoupled from ENaC function. The absence of benzamil-induced urinary alkalization in HKA double-knockout mice confirmed the direct involvement of these enzymes. The inhibitory effect of benzamil was also shown in vitro for the pig α -isoform of HKA. These results suggest a revised explanation of the benzamil effect on renal acid-base excretion. Considering the conditions used here, we suggest that it is caused by a direct inhibition of HKAs in the collecting duct and not by inhibition of the ENaC function. Bolus application of epithelial Na channel (EnaC) blockers causes marked and acute increases of urine pH. Here, we provide evidence that the underlying mechanism involves direct inhibition of the H -K pump in the collecting duct. This could provide a fundamental revision of the previously assumed mechanism that suggested a key role of ENaC inhibition in this response.