Effect of renal tubule-specific knockdown of the Na+/H+ exchanger NHE3 in Akita diabetic mice

Am J Physiol Renal Physiol. 2019 Aug 1;317(2):F419-F434. doi: 10.1152/ajprenal.00497.2018. Epub 2019 Jun 5.

Abstract

Na+/H+ exchanger isoform 3 (NHE3) contributes to Na+/bicarbonate reabsorption and ammonium secretion in early proximal tubules. To determine its role in the diabetic kidney, type 1 diabetic Akita mice with tubular NHE3 knockdown [Pax8-Cre; NHE3-knockout (KO) mice] were generated. NHE3-KO mice had higher urine pH, more bicarbonaturia, and compensating increases in renal mRNA expression for genes associated with generation of ammonium, bicarbonate, and glucose (phosphoenolpyruvate carboxykinase) in proximal tubules and H+ and ammonia secretion and glycolysis in distal tubules. This left blood pH and bicarbonate unaffected in nondiabetic and diabetic NHE3-KO versus wild-type mice but was associated with renal upregulation of proinflammatory markers. Higher renal phosphoenolpyruvate carboxykinase expression in NHE3-KO mice was associated with lower Na+-glucose cotransporter (SGLT)2 and higher SGLT1 expression, indicating a downward tubular shift in Na+ and glucose reabsorption. NHE3-KO was associated with lesser kidney weight and glomerular filtration rate (GFR) independent of diabetes and prevented diabetes-associated albuminuria. NHE3-KO, however, did not attenuate hyperglycemia or prevent diabetes from increasing kidney weight and GFR. Higher renal gluconeogenesis may explain similar hyperglycemia despite lower SGLT2 expression and higher glucosuria in diabetic NHE3-KO versus wild-type mice; stronger SGLT1 engagement could have affected kidney weight and GFR responses. Chronic kidney disease in humans is associated with reduced urinary excretion of metabolites of branched-chain amino acids and the tricarboxylic acid cycle, a pattern mimicked in diabetic wild-type mice. This pattern was reversed in nondiabetic NHE3-KO mice, possibly reflecting branched-chain amino acids use for ammoniagenesis and tricarboxylic acid cycle upregulation to support formation of ammonia, bicarbonate, and glucose in proximal tubule. NHE3-KO, however, did not prevent the diabetes-induced urinary downregulation in these metabolites.

Keywords: Na-glucose cotransporter 2; albuminuria; diabetes mellitus; diabetic nephropathy; hyperfiltration; proximal tubule; tubular growth.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Acid-Base Equilibrium* / genetics
  • Amino Acids, Branched-Chain / urine
  • Ammonia / urine
  • Animals
  • Bicarbonates / urine
  • Biomarkers / urine
  • Blood Glucose / metabolism
  • Diabetes Mellitus, Type 1 / genetics
  • Diabetes Mellitus, Type 1 / metabolism*
  • Diabetes Mellitus, Type 1 / physiopathology
  • Diabetes Mellitus, Type 1 / urine
  • Diabetic Nephropathies / genetics
  • Diabetic Nephropathies / metabolism*
  • Diabetic Nephropathies / physiopathology
  • Diabetic Nephropathies / urine
  • Disease Models, Animal
  • Energy Metabolism / genetics
  • Gene Expression Regulation
  • Gene Knockdown Techniques
  • Hydrogen-Ion Concentration
  • Kidney Tubules / metabolism*
  • Kidney Tubules / physiopathology
  • Male
  • Metabolomics / methods
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Renal Reabsorption*
  • Sodium / urine*
  • Sodium-Glucose Transporter 1 / genetics
  • Sodium-Glucose Transporter 1 / metabolism
  • Sodium-Glucose Transporter 2 / genetics
  • Sodium-Glucose Transporter 2 / metabolism
  • Sodium-Hydrogen Exchanger 3 / deficiency*
  • Sodium-Hydrogen Exchanger 3 / genetics

Substances

  • Amino Acids, Branched-Chain
  • Bicarbonates
  • Biomarkers
  • Blood Glucose
  • Slc5a1 protein, mouse
  • Slc5a2 protein, mouse
  • Slc9a3 protein, mouse
  • Sodium-Glucose Transporter 1
  • Sodium-Glucose Transporter 2
  • Sodium-Hydrogen Exchanger 3
  • Ammonia
  • Sodium