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Abstract: TH-PO840

DASH Diet Mechanism Dissected by Urinary Extracellular Vesicles

Session Information

Category: Health Maintenance‚ Nutrition‚ and Metabolism

  • 1400 Health Maintenance‚ Nutrition‚ and Metabolism


  • Bielopolski, Dana, The Rockefeller University, New York, New York, United States
  • Musante, Luca, The Pennsylvania State University - University Park Campus, University Park, Pennsylvania, United States
  • Barrows, Douglas, The Rockefeller University, New York, New York, United States
  • Carroll, Tom Samuel, The Rockefeller University, New York, New York, United States
  • Tobin, Jonathan N., The Rockefeller University, New York, New York, United States
  • Erdbruegger, Uta, University of Virginia, Charlottesville, Virginia, United States

The DASH (Dietary Approach to Stop Hypertension) diet is a proven intervention in lowering blood pressure, yet it’s mechanism has never been elucidated. We sought to characterize changes in urine exosome protein abundance pattern shifting from Western style diet to DASH diet.


9 volunteers were admitted for 14 days to the Rockefeller hospitalization unit, during which they transitioned from American style diet to DASH diet. blood and urine for electrolytes were collected daily and aldosterone was sampled four times.
Urine was centrifuged to discard cell debris at low speed and than the supernatant was centrifuged at 21,000g , the pellet Was termed P20. P20 was treated with TCEP to discard of Uromodulin. The supernatant was further centrifuged at 164,000g, and the pellet termed P100.P100 was fractioned with size exclusion chromatography to discard of Uromodulin, and the first two fractions that were positive for TSG101 and negative for Uromodulin were used for further analysis.


A total of 1,593 proteins were identified in P20+P100. 240 were uniquely expressed in P100 and 759 were uniquely expressed in P20. According to DAVID annotation tool both fractions were enriched for exosomes ( P20 with 95% and P100 with 97.4% of proteins involved P-value of 7.0E-51). Due to this differential expression we chose to unite the two fractions of P100 and P20 for mass spectrometry analysis of the DASH samples. 1800 proteins were identified and 226 crossed the significance threshold of change between trial days. 22 proteins were with upregulated and 25 were down regulated from day 0 to days 5 and 11. Both groups were enriched for extracellular Exosomes pathway
p-value= 3.8E-14 including SLC12A3 (NCC) in the upregulated and Aquaporine 2 (AQP2) in
the downregulated.


Following DASH diet implementation, the prevalence of NCC symporter increases and the prevalence of AQP2 decreases in uEV’s.
This leads to increased urine output and salt wasting similar to Gittelman syndrome.
prolonged thiazide treatment increases NCC abundance in urinary extracellular vesicles of essential hypertensive patients ,as well as in uEVs isolated from mice fed a high potassium diet. The increased abundance following the two stimuli may be a compensatory effect to counteract reduced NCC activity as a result of inhibition.


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