SUPPLEMENTARY DATA. Supplementary Table S1. Clinical characteristics of the study subjects.*
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1 Supplementary Table S1. Clinical characteristics of the study subjects.* T2D ND n (F/M) 66 (21/45) 25 (7/18) Age (years) 61.8 ± ± 7.3 # Body weight (kg) 95 ± ± 13 # Body mass index (kg. m -2 ) 31.6 ± ± 3.5 # HbA 1c (mmol/mol) 55.2 ± egfr (ml. min m 2 ) 86 ± ± 13 Fasting plasma glucose (mmol/l) 8.7 ± ± 0.4 # Fasting plasma insulin (pmol/l) 73 [74] 87 [50] Fasting plasma glucagon (pmol/l) 18 ± 8 21 ± 8 Meal glucose AUC (mmol/l) 317 [89] 211 [22] # Meal insulin AUC (nmol/l) 93 [65] 156 [123] # Meal glucagon AUC (nmol/ml) 1.07 ± ± 0.35 * Entries are mean±sd or median [interquartile range]; AUC = area-under-curve during 5 hours following a mixed meal; # p 0.05 vs T2D by Mann-Whitney test.
2 Supplementary Table S2. Effect of empagliflozin on renal glucose handling in nondiabetic subjects.* Baseline Acute Chronic # p 1 p 2 Fasting Plasma glucose (mmol. L -1 ) 5.8 ± 0.4 x 5.8 ± 0.4 x 5.5 ± 0.5 x ns 0.02 Urine output (ml. min -1 ) 3.7 ± ± ± 2.1 ns ns Creatinine clearance (ml. min ± 37 z 109 ± 31 x 110 ± 31 ns ns m -2 ) Glucose filtration rate 864 ± ± ± ns (µmol. min -1 ) x x y Glucose excretion rate 0.8 ± ± 74 x 172 ± 67 y < (µmol. min -1 ) Renal glucose clearance 0.2 ± ± ± 12 z < (ml. min -1 ) Fractional glucose excretion (%) 0.08 ± 23 ± 8 y 26 ± 11 z Meal Plasma glucose (mmol. L -1 ) 7.2 ± 0.7 a,x 6.6 ± 0.6 a,x 7.0 ± 0.4 b,x Urine output (ml. min -1 ) 2.9 ± ± 1.4 c 2.9 ± 1.3 ns 0.05 Creatinine clearance (ml. min ± 32 z 118 ± ± m -2 ) Glucose filtration rate 1185 ± 954 ± ± ns ns (µmol. min -1 ) 286 x x 308 c, y Glucose excretion rate 1.4 ± 1.9 x 388 ± ± 113 < (µmol. min -1 ) a,x c.y Renal glucose clearance 0.2 ± 0.2 y 59 ± 20 b 50 ± 15 c < (ml. min -1 ) Fractional glucose excretion (%) 0.09 ± 0.05 c, z 40 ± 7 a,y 35 ± 8 c * Entries are mean±sd; # IGT subjects only; p 1 = acute vs baseline, p 2 = chronic vs baseline (Wilcoxon sign rank); a p<0.0001; b p<0.001; c p<0.05 = Meal vs Fasting (Wilcoxon sign rank); x p<0.0001; y p<0.001; z p<0.05 = T2D vs NGT/IGT (Mann Whitney U test).
3 Supplementary Table S3. Effect of empagliflozin on renal solute handling in nondiabetic subjects.* Baseline Acute Chronic # p 1 p 3 Fasting ß-HB excretion rate (µmol. min -1 ) 0.07 [0.09] 0.14 [0.19] 0.19 [0.17] z ß-HB clearance rate (ml. min -1 ) 0.64 [0.83] 1.06 [0.56] 0.97 [1.13] ns ns ß-HB fractional excretion (%) 0.47 [0.64] 0.85 [0.84] z 0.62 [0.71] ns ns Lactate excretion rate (µmol. min -1 ) 0.06 [0.04] 0.15 [0.10] z 0.26 [0.17] < Lactate clearance rate (ml. min -1 ) 0.06 [0.04] 0.15 [0.11] 0.25 [0.17] Lactate fractional extraction 0.04 [0.04] 0.12 [0.14] 0.20 [0.05] Na + excretion rate (meq. min -1 ) 0.44 [0.24] 0.50 [0.26] 0.47 [0.20] ns ns Na + excretion (meq. 180min) 79 [43] 80 [42] 84 [35] ns ns Meal ß-HB excretion rate (µmol. min -1 ) 0.06 [0.03] c,z 0.11 [0.13] c,z 0.15 [0.19] c,z ß-HB clearance rate (ml. min -1 ) 0.87 [0.74] 1.31 [0.77] 1.04 [0.97] ns ns ß-HB fractional excretion (%) 0.55 [0.49] 0.85 [0.57] 0.72 [0.88] Lactate excretion rate (µmol. min -1 ) 0.06 [0.04] y 0.20 [0.20] b, z 0.33 [0.19] c < Lactate clearance rate (ml. min -1 ) 0.05 [0.03] 0.21 [0.17] 0.36 [0.26] Lactate fractional extraction (%) 0.03 [0.02] c,y 0.12 [0.15] z 0.27 [0.16] c Na + excretion rate (meq. min -1 ) 0.32 [0.14] c 0.42 [0.13] c 0.39 [0.19] Na + excretion (meq. 300min) 96 [42] 126 [39] c 117 [57] c * entries are median [IQR]; # IGT subjects only; p 1 = acute vs baseline, p 2 = chronic vs baseline; a p<0.0001; b p<0.001; c p<0.05 = Meal vs Fasting; x p<0.0001; y p<0.001; z p<0.05 = T2D vs NGT/IGT (Mann Whitney U test).
4 Supplementary Table S4. Effect of empagliflozin on laboratory parameters.* Baseline Acute Chronic # p 1 p 2 T2D HbA 1c (%) 7.20 ± ± ± 0.53 ns < Hematocrit (%) 40.8 ± ± ± ns Red blood cells ( L -1 ) 4.46 ± ± ± 0.40 < Hemoglobin (g/l) 139 ± ± ± 12 < White blood cells ( L -1 ) 6.2 ± ± ± 1.6 ns ns Fasting plasma albumin (g. L -1 ) 41.2 ± ± ± 2.6 ns 0.01 Meal plasma albumin (g. L -1 ) 38.5 ± 2.4 a 39.6 ± 2.6 a 39.7 ± 2.8 a < < Systolic blood pressure (mmhg) 138 ± ± ± Serum NT-proBNP (ng/l) 41 [43] - 40 [48] ns ns ND HbA 1c (%) 5.48 ± 0.38 x 5.50 ± 0.35 x 5.44 ± 0.42 x ns ns Hematocrit (%) 42.8 ± 2.9 z 42.4 ± 3.1 z 42.6 ± 3.0 ns ns Red blood cells ( L -1 ) 4.62 ± ± 0.31 z 4.49 ± ns Hemoglobin (g. L -1 ) 143 ± ± 10 z 143 ± ns White blood cells ( L -1 ) 6.6 ± ± ± 1.7 ns ns Fasting plasma albumin (g. L -1 ) 40.0 ± ± ± 1.7 z 0.02 ns Meal plasma albumin (g. L -1 ) 38.2 ± 1.9 c 39.3 ± 2.1 b 38.8 ± 1.8 b < Systolic blood pressure (mmhg) 130 ± 11 z 129 ± 12 z 132 ± 15 ns ns Serum NT-proBNP (ng/l) 39 [81] - 41 [70] ns ns * entries are mean ± SD or median [IQR]; # Among ND includes IGT subjects only; p 1 = acute vs baseline, p 2 = chronic vs baseline; a p<0.0001; b p<0.001; c p<0.05 = Meal vs Fasting; x p<0.0001; y p<0.001; z p<0.05 = T2D vs NGT/IGT (Mann Whitney U test).
5 Supplementary Figure S1. Urine output (top) and creatinine clearance (bottom) during fasting and following meal ingestion at baseline, after acute and chronic empagliflozin administration in patients with type 2 diabetes. Plots are mean SEM. * p 0.05 vs Baseline; p 0.05 vs Fasting.
6 Supplementary Figure S2. Time-course of urinary excretion of s-hydroxybutyrate (s-hb), lactatae, and sodium in T2D patients at baseline and following acute and chronic administration of 25 mg empagliflozin.
7 Supplementary Figure S3. Relationship between glucose excretion and urinary excretion of s- hydroxybutyrate top), lactate (middle), and sodium (bottom) during chronic treatment with empagliflozin. Data from fasting and postmeal urine collections from T2D patients and nondiabetic subjects are plotted together. Best fit (full lines) and 95% confidence intervals (dotted line) are shown along with correlation coefficients and their p values.
8 Supplementary Figure S4. Fasting serum erythropoietin concentrations at baseline and after 4 weeks of treatment with 25 mg empagliflozin. Data are median and interquartile range. * p= for the difference between baseline and follow up by 2-way ANOVA for repeated measures.
9 Supplementary Figure S5. Relationship between serum erythropoietin concentrations (EPO) and blood hemoglobin concentrations at baseline (top), and after 4 weeks of treatment with 25 mg empagliflozin (bottom). Best fit (full lines) and 95% confidence intervals (dotted line) are shown along with correlation coefficients and their p values.
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