High-glucose-triggered apoptosis in cultured endothelial cells

• Published in: Diabetes (New York, N.Y.) Vol. 44; no. 11; pp. 1323 - 1327
• Main Authors: BAUMGARTNER-PARZER, S. M, WAGNER, L, PETTERMANN, M, GRILLARI, J, GESSL, A, WALDHÄUSL, W
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Diabetes Association 01-11-1995
• Subjects: Ageing, cell death; Apoptosis - drug effects; Biological and medical sciences; Blotting, Northern; Cell death; Cell physiology; Cells, Cultured; Clusterin; Cytology; Dextrose; DNA - drug effects; DNA - isolation & purification; DNA - metabolism; Dose-Response Relationship, Drug; Electrophoresis, Agar Gel; Endothelium; Endothelium, Vascular - cytology; Endothelium, Vascular - drug effects; Endothelium, Vascular - physiology; Flow Cytometry; Fundamental and applied biological sciences. Psychology; Gene Expression - drug effects; Glucose; Glucose - pharmacology; Glycoproteins - biosynthesis; Humans; Kinetics; Molecular and cellular biology; Molecular Chaperones; Physiological aspects; RNA, Messenger - analysis; RNA, Messenger - biosynthesis; Thymidine - metabolism; Umbilical Veins; Human; Endothelial cell; Blood vessel; Circulatory system; Glucose; In vitro; Apoptosis
• ISSN: 0012-1797; 1939-327X
• DOI: 10.2337/diab.44.11.1323

High ambient glucose concentration, linked to vascular complications in diabetes in vivo, modulates mRNA expression of fibronectin, collagen, tissue-type plasminogen activator, and plasminogen activator inhibitor and induces delayed replication and excess cell death in cultured vascular endothelial cells. To determine the role of high ambient glucose (30 mmol/l) in apoptosis, paired cultures of individual isolates of human umbilical vein endothelial cells (HUVECs) were exposed to both high (30 mmol/l) and low (5 mmol/l) concentrations of glucose for short-term (24, 48, and 72 h) and long-term (13 +/- 1 days) experiments. Incubation of HUVECs with high glucose for > 48 h increased DNA fragmentation (13.7 +/- 6.5% of total DNA, mean +/- SD) versus cultures kept in 5 mmol/l glucose (10.9 +/- 5.6%, P < 0.005), as measured by [3H]thymidine assays. Data were confirmed by apoptosis-specific fluorescence-activated cell sorter analysis of confluent HUVEC cultures, which displayed after long-term exposure to 30 mmol/l glucose a 1.5-fold higher prevalence of apoptosis than control cultures exposed to 5 mmol/l glucose (P < 0.005). In contrast, no increase in DNA fragmentation in response to 30 mmol/l glucose was seen for standardized cell lines (K 562, P 815, YT) and fibroblasts. Expression of clusterin mRNA, originally reported to be a molecular marker of apoptosis, was only slightly affected by short-term (24-h) high-glucose exposure but was significantly reduced after long-term incubation in 30 mmol/l glucose (82.2 +/- 13.8% of control) versus 5 mmol/l glucose, which questions the role of clusterin gene expression as a marker of apoptosis.