The Dietary Approach to Stop Hypertension (DASH) diet is a proven intervention to treat hypertension, yet its mechanism is not clearly known. We investigated the change in protein abundance patterns in urine extracellular vesicles (uEVs) following DASH diet implementation. A pilot study was carried out to compare uEVs isolated using three different methods: a low centrifugation (P20), high centrifugation (P100), and a combination of both (P20 and P100). Uromodulin was removed by size exclusion chromatography and low ionic strength washing. Mass spectrometry analysis identified 1,593 proteins in the combined fraction (P20+P100), 1434 in the P20 fraction and 1229 in the P100 fraction. The combined fraction was chosen for further analysis. Statistical analysis was carried out using R and Limma to identify all proteins that changed before and during 11 days of DASH intervention (p < 0.05) as well as between individual timepoints. Nine hypertensive volunteers were admitted for a 14-day supervised transition from American style diet to DASH diet. First-void urine was collected on days 0, 5, 11 for uEV processing. In total, 1800 proteins were identified across all 27 DASH samples with 22 proteins upregulated and 25 down regulated between day 0 and both days 5 and 11. These included increased abundance of SLC12A3 (NCC) and reduced abundance of Aquaporine 2. These changes could explain the increased urine volume and reduced sodium reabsorption that lead to blood pressure reduction following consumption of the high potassium and low sodium DASH diet. uEVs may serve as a surrogate to a more invasive procedure.

The authors have declared no competing interest.

NCT04142138

This work was supported in part by the National Center for Advancing Translational Sciences, National Institutes of Health, through Rockefeller University, Grant #UL1TR001866. This work was supported by the Young Investigator Grant of the National Kidney Foundation

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