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Advanced glycation end products (AGEs) are non-enzymatic glycations of protein or lipids when exposed to sugars. AGEs are involved in several systemic conditions such as diabetes mellitus or ageing and in organ dysfunction of the brain (neurodegeneration), heart (heart failure), and kidney (chronic kidney disease). Indeed, the receptor of AGEs (RAGE) is highly expressed in the brain, kidneys, and lungs. Several cell types also express RAGE, including endothelial cells.
In the kidney, AGEs have been implicated in diabetic nephropathy, hypertensive nephropathy, and IgA Nephropathy. The extent of glycation is observed to be correlated with the fibrosis of the kidney.
All the conditions modified by AGEs share the presence of Endothelial dysfunction (ED), and therefore we hypothesize that AGEs might damage the kidney by acting on endothelial cells.
Furthermore, the recent introduction of SGLT2 (sodium-glucose cotransporter 2) inhibitors (SGLT2i) appears to be beneficial in all these conditions, and therefore it is plausible they act, at least in part, on ED.
Aim
We have investigated if kidney protection by SGLT2i might be mediated by endothelial cells, through a scavenger activity on AGEs.
Methods
We studied the effects of empagliflozin on the human endothelial cell line (EA. hy926) in the presence of Glycated Human Serum Albumin. Multiple approaches were employed to evaluate the effect of Empagliflozin (SGLT2i) on AGEs-induced ED, such as cell viability, evaluation of ROS production, and cell-morphological parameters by phase contrast optical microscopy. The treatment is done with two concentrations of Glycated albumin and two concentrations of Empagliflozin based on IC50. Cell morphology has also been analyzed using Atomic Force Microscopy and expansion microscopy.
The results suggest that SGLT2i improve cell vitality at baseline and protects endothelial cells from AGEs-induced cytotoxicity. It was observed that Empagliflozin was able to increase the cell vitality up to 35% with respect to AGEs and provide protection from AGEs. It was also observed that there is an approximately 55-60 % reduction in ROS production in the endothelial cells. The observed protective effect of Empagliflozin towards AGEs is both dose and time-dependent. Moreover, A new protocol for studying cell morphology has been optimized via Expansion microscopy which allows cells to expand, thus facilitating the visualization of subcellular organelle under a simple microscope. This new tool is demonstrated to help study cell morphology and Endothelial dysfunction.
Our data suggest that SGLT2i might be beneficial through protection from AGEs; this also suggests that they might prove beneficial in the ageing process and other vascular diseases.