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  • Advanced glycation end products‑induced mitochondrial energy metabolism dysfunction alters proliferation of human umbilical vein endothelial cells.

Advanced glycation end products‑induced mitochondrial energy metabolism dysfunction alters proliferation of human umbilical vein endothelial cells.

Molecular medicine reports (2017-04-28)
Yuan Li, Ye Chang, Ning Ye, Yintao Chen, Naijin Zhang, Yingxian Sun
ABSTRACT

Advanced glycation end products (AGEs) restrain the proliferation of endothelial cells, which is an important determinant of diabetic vasculopathy. Mitochondrial biogenesis serves an essential role in cellular adaptation and repair. The current study aimed to investigate alterations in mitochondrial energy metabolism in human umbilical vein endothelial cells (HUVECs) and the latent mechanism regulated by AGEs. The proliferation of cultured HUVECs stimulated with AGEs was detected using an MTT assay and a real‑time cell analyzer (RTCA). Mitochondrial energy metabolism was measured using a Seahorse metabolic flux analyzer. Mitochondrial membrane potential was detected under fluorescence microscopy following staining with tetraethylrhodamine and MitoTracker Red. Respiratory chain complexes I‑V were detected using western blotting. MTT and RTCA assays demonstrated that AGEs treatment significantly inhibited the viability and proliferation of HUVECs when compared with bovine serum albumin treatment. Results from the Seahorse metabolic flux analyzer indicated that mitochondrial aerobic respiration and glycolysis declined following AGEs treatment. In addition, mitochondrial membrane potential and the expression of mitochondrial respiration chain complexes I/II/III/IV/V notably decreased in the presence of AGEs. In conclusion, the results of the present study indicated that AGEs exhibited an inhibitory effect on the proliferation in HUVECs potentially by mediating the dysfunction of mitochondrial energy metabolism and glycolysis. This may provide a new consideration for therapeutic methods in diabetic vascular complications.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Basal Medium Eagle, With Earle′s salts and L-glutamine, without sodium bicarbonate, powder, suitable for cell culture
Sigma-Aldrich
Anti-GAPDH Antibody, from chicken, purified by affinity chromatography
Sigma-Aldrich
Goat Anti-Rabbit IgG Antibody, Rhodamine conjugate, Species Adsorbed, 1.5 mg/mL, Chemicon®
Millipore
Goat Anti-Mouse IgG Antibody, Rhodamine conjugate, Species Adsorbed, 1.4 mg/mL, Chemicon®
Sigma-Aldrich
Human Umbilical Vein Endothelial Cells: HUVEC: Pre-Screened for Angiogenesis & VEGF signaling, neonatal