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Metformin is the first-line drug in the treatment of type 2 diabetes mellitus. The aim of this work was to evaluate the efficacy of metformin treatment in reducing blood pressure and investigate the molecular mechanism using a preclinical animal model. Adult male and female diabetic db/db mice with a blood glucose of greater than 300 mg/dl were salt-loaded (8% NaCl) for 10 days to induce hypertension. The mice were subject to metabolic cage studies for 24 hour urine collections in order to measure urinary electrolytes, albumin, and creatinine. Blood pressure was measured weekly by the tail-cuff method to assess the effect of metformin or vehicle given by oral gavage (dose of 60 mg/kg of body weight per day). At the end of the study the mice was euthanized and the left kidney was formalin-fixed and paraffin-embedded for immunohistochemistry while the right kidney was homogenized for Western blotting. Western blotting showed attenuation of total NCC and phospho-NCC in diabetic db/db mice given an oral gavage of metformin (Pearson correlation coefficient: 0.9470 +/- 2.52e-3) compared to vehicle (Pearson correlation coefficient: 0.9800 +/- 2.86e-3). Immunohistochemical analysis showed less co-localization of the actin cytoskeleton protein filamin and phosphorylated NCC in the metformin treated group compared to the control group. Taken together, we show metformin decreases sodium retention and blood pressure by reducing the density of renal NCC at the luminal membrane and the association between NCC and the actin cytoskeleton.

journal of the endocrine society

SAT-LB126 Metformin Attenuates Sodium Retention and Blood Pressure in Hypertensive Diabetic Mice by Reducing the Phosphorylation of Renal NCC and Its Association With the Actin Cytoskeleton