Phosphodiesterase 5A Inhibition Decreases NHE-1 Activity Without Altering Steady State pHi: Role of Phosphatases

Cellular Physiology and Biochemistry

关键词: Phosphodiesterase 5A;    NHE-1;    Protein kinase G;    Protein phosphatase 2A;   

DOI  :  10.1159/000322321

学科分类：分子生物学，细胞生物学和基因

来源: S Karger AG

【 摘 要 】

Background/Aims This study aimed to identify the signaling pathway for the proposed link between phosphodiesterase-5A (PDE5A) inhibition and decreased cardiac Na⁺/H⁺ exchanger (NHE-1) activity. Methods NHE-1 activity was assessed in rat isolated papillary muscles by the Na⁺-dependent initial pH_i recovery from a sustained acidosis (ammonium prepulse). ERK1/2, p90RSK and NHE-1 phosphorylation state during acidosis was determined. Results PDE5A inhibition (1 µmol/L sildenafil, SIL) did not modify basal pH_i but significantly blunted pH_i recovery after sustained acidosis. Although preventing ERK1/2- p90RSK signaling pathway (10 µmol/L U0126) mimicked SIL effect, SIL did not blunt the acidosis-mediated increase in kinases activation. SIL+U0126 did not show additive effect on NHE-1 activity. Then, we hypothesized that SIL could be activating phophasatases (PP1 and/or PP2A) to directly dephosphorylate NHE-1 despite preserved ERK1/2-p90RSK activation. Non-specific phosphatases inhibition (1 µmol/L okadaic acid) canceled SIL effect on pH_i recovery from acidosis. Same result was observed by inhibiting PP2A either with a lower dose of okadaic acid (1 nmol/L) or, more specifically, with 100 µmol/L endothall. Consistently, NHE-1 phosphorylation at Ser703 increased after acidosis, SIL prevented this effect and PP2A inhibition (endothall) reverted SIL effect. Conclusion We suggest that PDE5A inhibitors decrease NHE-1 phosphorylation and activity through a mechanism that involves PP2A activation.

【 授权许可】

CC BY-NC-ND