(1) Background: empagliflozin, sodium-glucose co-transporter 2 (SGLT-2) inhibitor, is an effective antidiabetic agent with strong cardio- and nephroprotective properties. The mechanisms behind its cardio- and nephroprotection are still not fully clarified. (2) Methods: we used male hereditary hypertriglyceridemic (hHTG) rats, a non-obese model of dyslipidaemia, insulin resistance, and endothelial dysfunction fed standard diet with or without empagliflozin for six weeks to explore the molecular mechanisms of empagliflozin effects. Nuclear magnetic resonance (NMR)-based metabolomics; quantitative PCR of relevant genes involved in lipid and glucose metabolism, or senescence; glucose and palmitic acid oxidation in isolated tissues and cell lines of adipocytes and hepatocytes were used. (3) Results: empagliflozin inhibited weight gain and decreased adipose tissue weight, fasting blood glucose, and triglycerides and increased HDL-cholesterol. It also improved insulin sensitivity in white fat. NMR spectroscopy identified higher plasma concentrations of ketone bodies, ketogenic amino acid leucine and decreased levels of pyruvate and alanine. In the liver, adipose tissue and kidney, empagliflozin up-regulated expression of genes involved in gluconeogenesis and down-regulated expression of genes involved in lipogenesis along with reduction of markers of inflammation, oxidative stress and cell senescence. (4) Conclusion: multiple positive effects of empagliflozin, including reduced cell senescence and oxidative stress, could contribute to its long-term cardio- and nephroprotective actions.

Cardiometabolic Research Division Centre for Experimental Medicine Institute for Clinical and Experimental Medicine 140 21 Prague Czech Republic

Department of Analytical Chemistry Faculty of Science Palacky University Olomouc 779 00 Olomouc Czech Republic

Diabetes Centre Institute for Clinical and Experimental Medicine 140 21 Prague Czech Republic

Institute of Medical Biochemistry and Laboratory Diagnostics 1st Faculty of Medicine Charles University and General University Hospital 128 08 Prague Czech Republic

Laboratory of Animal Biochemistry Department of Biochemistry and Microbiology University of Chemistry and Technology Prague 166 28 Prague Czech Republic

Laboratory of Molecular Structure Characterization Institute of Microbiology Czech Academy of Sciences 142 20 Prague Czech Republic

Laboratory of Molecular Therapy Institute of Biotechnology Czech Academy of Sciences 252 50 Prague Czech Republic

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Sodium Glucose Cotransporter-2 Inhibitors: Spotlight on Favorable Effects on Clinical Outcomes beyond Diabetes