Metabolomic profiling in kidney cells treated with a sodium glucose-cotransporter 2 inhibitor

Abstract

We aimed to determine the metabolomic profile of kidney cells under high glucose conditions and following sodium-glucose cotransporter 2 (SGLT2) inhibitor treatment. Targeted metabolomics using the Absolute IDQ-p180 kit was applied to quantify metabolites in kidney cells stimulated with high glucose (25 and 50 mM) and treated with SGLT2 inhibitor, dapagliflozin (2 µM). Primary cultured human tubular epithelial cells and podocytes were used to identify the metabolomic profile in high glucose conditions following dapagliflozin treatment. The levels of asparagine, PC ae C34:1, and PC ae C36:2 were elevated in tubular epithelial cells stimulated with 50 mM glucose and were significantly decreased after 2 µM dapagliflozin treatment. The level of PC aa C32:0 was significantly decreased after 50 mM glucose treatment compared with the control, and its level was significantly increased after dapagliflozin treatment in podocytes. The metabolism of glutathione, asparagine and proline was significantly changed in tubular epithelial cells under high-glucose stimulation. And the pathway analysis showed that aminoacyl-tRNA biosynthesis, arginine and proline metabolism, glutathione metabolism, valine, leucine and isoleucine biosynthesis, phenylalanine, tyrosine, and tryptophan biosynthesis, beta-alanine metabolism, phenylalanine metabolism, arginine biosynthesis, alanine, aspartate and glutamate metabolism, glycine, serine and threonine metabolism were altered in tubular epithelial cells after dapagliflozin treatment following 50 mM glucose compared to those treated with 50 mM glucose.

Role of the CCL20/CCR6 axis in tubular epithelial cell injury: Kidney‐specific translational insights from acute kidney injury to chronic kidney disease

“…The methods for establishing primary proximal tubule cell cultures have been described previously 21,22,29,30 . In brief, the kidney cortex was carefully sectioned, and the tissue pieces were treated with Hank's balanced salt solution containing 3 mg/mL collagenase (Sigma‐Aldrich, St. Louis, MO, USA) and incubated at 37°C for 1 h. Next, the cells were rinsed with phosphate‐buffered saline (PBS) through a series of sieves and then spun down at 500× g for 5 min.…”

“…The methods for establishing primary proximal tubule cell cultures have been described previously. 21,22,29,30 In brief, the kidney cortex was carefully sectioned, and the tissue pieces were treated with Hank's balanced salt solution containing 3 mg/mL collagenase (Sigma-Aldrich, St. Louis, MO, USA) and incubated at 37°C for 1 h. Next, the cells were rinsed with phosphate-buffered saline (PBS) through a series of sieves and then spun down at 500× g for 5 min. Following this, the cells were treated with DMEM/F12 media (Lonza) for 4-6 h, after which the tubular cells floating in the media were harvested and grown on collagen-coated plates (BD Biosciences) until colonies of epithelial cells formed.…”