A unifying model of glucotoxicity in human renal proximal tubular epithelial cells and the effect of the SGLT2 inhibitor dapagliflozin

Abstract

Background: Glucotoxicity in renal tubular epithelial cells (RPTECs) contributes to the pathogenesis of diabetic nephropathy. Sodium-glucose cotransporter 2 (SGLT2) inhibitors may exert their renoprotective effect by preventing glucotoxicity. We tested whether the confirmed in capillary endothelial cells unifying model of glucotoxicity can be applied in RPTECs and the impact of dapagliflozin. Methods: In primary human RPTECs cultured in normal or high glucose medium in the presence or not of dapagliflozin, we assessed glucose consumption, SCLT2 expression, reactive oxygen species (ROS) production, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) activity, d-sorbitol and methylglyoxal cell content, O-linked β-N-acetyl glucosamine (O-Glc-NAc)-modified proteins, protein kinase C (PKC) activity, transforming growth factor-β1 (TGF-β1), interleukin-8 (IL-8), cell necrosis, and cell apoptosis using colorimetric and immunoenzymatic assays, and western blotting. Results: High glucose increases SGLT2 expression and glucose consumption. ROS are overproduced, and GAPDH is inhibited. The accumulation due to GAPDH inhibition glycolytic products are diverted into four noxious pathways. The polyol pathway assessed by d-sorbitol, the hexosamine pathway determined by O-GlcNAc-modified proteins, the lipid synthesis pathway assessed by PKC activity, and the advanced glycation end-products (AGEs) formation assessed by methylglyoxal. Eventually, these paths lead to overproduction of TGF-β1 and IL-8, as well as to cell necrosis and apoptosis. Dapagliflozin ameliorates all the above cascade of events. Conclusions: Our results support a unifying model for glucotoxicity in RPTECs. Dapagliflozin by decreasing the elevated glucose influx into the RPTECs under high glucose conditions ameliorates glucotoxicity. © 2020, Springer Nature B.V.