Uraemic plasma decreases the expression of ABCA1, ABCG1 and cell-cycle genes in human coronary arterial endothelial cells

• Published in: Nephrology, dialysis, transplantation Vol. 22; no. 2; pp. 409 - 416
• Main Authors: Cardinal, Héloise, Raymond, Marc-André, Hébert, Marie-Josée, Madore, François
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.02.2007; Oxford Publishing Limited (England)
• Subjects: Aged; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; apoptosis; Apoptosis - genetics; atherosclerosis; ATP Binding Cassette Transporter 1; ATP Binding Cassette Transporter, Sub-Family G, Member 1; ATP-Binding Cassette Transporters - biosynthesis; ATP-Binding Cassette Transporters - genetics; Biological and medical sciences; Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis; Cell Cycle - physiology; Cell Proliferation; Cells, Cultured; Coronary Vessels - metabolism; Coronary Vessels - pathology; Emergency and intensive care: renal failure. Dialysis management; endothelial cells; Endothelium, Vascular - metabolism; Female; gene expression; Gene Expression - physiology; Humans; Intensive care medicine; Male; Medical sciences; Microarray Analysis; Middle Aged; Nephrology. Urinary tract diseases; Nephropathies. Renovascular diseases. Renal failure; Plasma; Polymerase Chain Reaction; Renal failure; RNA - genetics; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; uraemia; Uremia - blood; Uremia - pathology; atherosclerosis; apoptosis; endothelial cells; cell proliferation; uraemia; gene expression; Kidney disease; Human; Cell proliferation; Endothelial cell; Urinary system disease; Hemodialysis; Cardiovascular disease; Gene expression; Vascular disease; Extrarenal dialysis; Atherosclerosis; Cell cycle; Renal failure; Apoptosis
• ISSN: 0931-0509; 1460-2385
• DOI: 10.1093/ndt/gfl619

Background. Uraemia is associated with endothelial dysfunction, but the effect of uraemic plasma on the gene expression pattern of human coronary arterial endothelial cells (HCAEC) has never been defined. Methods. HCAECs were exposed for 48 h to a culture medium supplemented with 20% uraemic vs normal plasma. We extracted mRNA and hybridized it onto Affymetrix HG-U133 Plus2 microarrays. We validated our findings for five genes of interest by real-time PCR and performed evaluations of cell proliferation and apoptosis in HCAECs exposed to uraemic vs normal plasma. Results. Six genes involved in the regulation of cell-cycle progression (CDK-1, topoisomerase II, PDZ-binding kinase, CDCA1, protein SDP35, E2F transcription factor 8) and two genes of the cholesterol efflux system (ABCA1 and ABCG1) were down-regulated in HCAECs exposed to uraemic plasma (>1.75-fold change vs normal). Real-time PCR confirmed the down-regulation observed in the microarray experiment. Cell proliferation was significantly decreased in HCAECs exposed to uraemic vs normal plasma for 48 h (86 vs 95% of serum-starved control, P = 0.006). Exposure to uraemic plasma for 48 h was associated with increased apoptosis of HCAEC as compared with normal plasma (7.7 vs 2.8%, P < 0.001), a phenomenon that was further enhanced when oxidized LDLs (150 µg protein/ml) were added to the medium containing uraemic plasma (16.9 vs 7.7%, P < 0.001). Conclusions. The down-regulation of genes involved in cell-cycle progression and cholesterol efflux from HCAECs exposed to uraemic conditions could contribute to enhancing endothelial dysfunction and atherosclerosis in patients with chronic renal failure.