A Non-Targeted Capillary Electrophoresis-Mass Spectrometry Strategy to Study Metabolic Differences in an In Vitro Model of High-Glucose Induced Changes in Human Proximal Tubular HK-2 Cells

Abstract

Diabetic nephropathy is characterized by the chronic loss of kidney function due to high glucose renal levels. HK-2 proximal tubular cells are good candidates to study this disease. The aim of this work was to study an in vitro model of high glucose-induced metabolic alterations in HK-2 cells to contribute to the pathogenesis of this diabetic complication. An untargeted metabolomics strategy based on CE-MS was developed to find metabolites affected under high glucose conditions. Intracellular and extracellular fluids from HK-2 cells treated with 25 mM glucose (high glucose group), with 5.5 mM glucose (normal glucose group), and with 5.5 mM glucose and 19.5 mM mannitol (osmotic control group) were analyzed. The main changes induced by high glucose were found in the extracellular medium where increased levels of four amino acids were detected. Three of them (alanine, proline, and glutamic acid) were exported from HK-2 cells to the extracellular medium. Other affected metabolites include Amadori products and cysteine, which are more likely cause and consequence, respectively, of the oxidative stress induced by high glucose in HK-2 cells. The developed CE-MS platform provides valuable insight into high glucose-induced metabolic alterations in proximal tubular cells and allows identifying discriminative molecules of diabetic nephropathy.

Long Non-Coding RNA Small Nucleolar RNA Host Gene 5 (SNHG5) Regulates Renal Tubular Damage in Diabetic Nephropathy via Targeting MiR-26a-5p

“…Proximal renal tubules are the main driving force of renal pathology in DM [22]. Therefore, HK2 cell line is a good candidate for studying how high glucose induction promotes the occurrence and progression of DN [23]. Growing evidence suggested that the pathogenesis of DN is related to dysfunction of renal proximal tubular epithelial cell, oxidative stress, inflammation, and fibrosis under a high-glucose condition [24].…”

Time-Dependent Influence of High Glucose Environment on the Metabolism of Neuronal Immortalized Cells

“…The effects of a high glucose environment on metabolome have been previously reported for cell lines from pancreas (BRIN-BD11 and INS-1E), [27] liver (HepG2) [28] and kidneys (HK-2). [29][30][31] In the studies mentioned, 25 mM was the maximum concentration of glucose employed to treat the cells, respect to a physiological glucose concentration equal to 5.5 mM. [32] Due to the serious neurological complications associated with pathologies as diabetes, we propose here the HN9.10e hippocampal neuroblasts as model line to study hyperglycemia metabolic effects.…”

Piceatannol Protects against High Glucose-Induced Injury of Renal Tubular Epithelial Cells via Regulating Carbonic Anhydrase 2

“…For subsequent studies, HK-2 cells were divided into these groups as needed: control group (HK-2 cells were treated with 5.5 mmol/L D-glucose [G8150; Solarbio, China] for 48 h), osmotic control (Os-control) group (HK-2 cells were treated with 5.5 mmol/L D-glucose and 19.5 mM mannitol (SM8120; Solarbio) for 48 h) [21], PIC-H group (HK-2 cells were treated with 20 μmol/L PIC [SP9130; Solarbio, China] for 24 h, and 5.5 mmol/L D-glucose for 48 h), HG group (HK-2 cells were treated with 30 mmol/L D-glucose for 48 h), HG+PIC-L group (HK-2 cells were first treated with 10 μmol/L PIC for 24 h and then with 30 mmol/L D-glucose for 48 h), and HG+PIC-H group (HK-2 cells were treated with 20 μmol/L PIC for 24 h and 30 mmol/L D-glucose for 48 h).…”