Extracellular acidosis and very low [Na⁺ ] inhibit NBCn1- and NHE1-mediated net acid extrusion from mouse vascular smooth muscle cells

Aim: The electroneutral Na⁺ , HCO3- cotransporter NBCn1 and Na⁺ /H⁺ exchanger NHE1 regulate acid-base balance in vascular smooth muscle cells (VSMCs) and modify artery function and structure. Pathological conditions - notably ischaemia - can dramatically perturb intracellular (i) and extracellular (o) pH and [Na⁺ ]. We examined effects of low [Na⁺ ]_o and pH_o on NBCn1 and NHE1 activity in VSMCs of small arteries.

Methods: We measured pH_i by 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein-based fluorescence microscopy of mouse mesenteric arteries and induced intracellular acidification by NH4+ prepulse technique.

Results: NBCn1 activity - defined as Na⁺ -dependent, amiloride-insensitive net base uptake with CO₂ /HCO3- present - was inhibited equally when pH_o decreased from 7.4 (22 mm HCO3-/5% CO₂ ) by metabolic (pH_o 7.1/11 mm HCO3-: 22 ± 8%; pH_o 6.8/5.5 mm HCO3-: 61 ± 7%) or respiratory (pH_o 7.1/10% CO₂ : 35 ± 11%; pH_o 6.8/20% CO₂ : 56 ± 7%) acidosis. Extracellular acidosis more prominently inhibited NHE1 activity - defined as Na⁺ -dependent net acid extrusion without CO₂ /HCO3- present - at both pH_o 7.1 (45 ± 9%) and 6.8 (85 ± 5%). Independently of pH_o , lowering [Na⁺ ]_o from 140 to 70 mm reduced NBCn1 and NHE1 activity <20% whereas transport activities declined markedly (25-50%) when [Na⁺ ]_o was reduced to 35 mm. Steady-state pH_i decreased more during respiratory (ΔpH_i /ΔpH_o = 71 ± 4%) than metabolic (ΔpH_i /ΔpH_o = 30 ± 7%) acidosis.

Conclusion: Extracellular acidification inhibits NBCn1 and NHE1 activity in VSMCs. NBCn1 is equivalently inhibited when pCO₂ is raised or [HCO3-]_o decreased. Lowering [Na⁺ ]_o inhibits NBCn1 and NHE1 markedly only below the typical physiological and pathophysiological range. We propose that inhibition of Na⁺ -dependent net acid extrusion at low pH_o protects against cellular Na⁺ overload at the cost of intracellular acidification.