Endothelial pro-atherosclerotic response to extracellular diabetic-like environment: possible role of thioredoxin-interacting protein.
BACKGROUND. High blood and tissue concentrations of glucose and advanced glycation end-products (AGEs) are thought to play an important role in the development of vascular diabetic complications. Therefore, the impact of extracellular AGEs and different glucose concentrations was evaluated by studying the gene expressions and the underlying cellular pathways involved in the development of inflammatory pro-atherosclerotic processes observed in cultured endothelial cells. METHODS. Fresh human umbilical vein cord endothelial cells (HUVEC) were treated in the presence of elevated extracellular glucose concentrations (5.5-28 mmol/l) with and without AGE-human serum albumin (HSA). Affymetrix GeneChip(R) Human Gene 1.0 ST arrays were used for gene expression analysis (total 20 chips). Genes of interest were further validated using real-time PCR and western blot techniques. RESULTS. Microarray analysis revealed significant changes in some gene expressions in the presence of the different stimuli, suggesting that different pathways are involved. Six genes were selected for validation as follows: thioredoxin-interacting protein (TXNIP), thioredoxin (TXN), nuclear factor of kappa B (NF-kappaB), interleukin 6 (IL6), interleukin 8 (IL8) and receptor of advanced glycation end-products (RAGE). Interestingly, it was found that the association of AGEs together with the highest pathophysiological concentration of glucose (28 mmol/l) diminished the expression of these specific genes, excluding TXN. CONCLUSIONS. In the present model that mimics a diabetic environment, the relatively short-term experimental conditions used showed an unexpected blunting action of AGEs in the presence of the highest glucose concentration (28 mmol/l). The interactive cellular pathways involved in these processes should be further investigated.
Zitman-Gal
Tali
T
Renal Physiology Laboratory, Department of Nephrology and Hypertension, Meir Medical Center, Kfar Saba, Israel. tali.gal@clalit.org.il
Green
Janice
J
Pasmanik-Chor
Metsada
M
Oron-Karni
Varda
V
Bernheim
Jacques
J
D050197:Atherosclerosis / Q000503:physiopathology*; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology*; D002478:Cells, Cultured; D048909:Diabetes Complications / Q000503:physiopathology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D005109:Extracellular Matrix / Q000502:physiology*; D005947:Glucose / Q000378:metabolism; D017127:Glycation End Products, Advanced / Q000378:metabolism; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics / Q000502:physiology; D016209:Interleukin-8 / Q000235:genetics / Q000502:physiology; D016328:NF-kappa B / Q000235:genetics / Q000502:physiology; D020411:Oligonucleotide Array Sequence Analysis; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic / Q000235:genetics / Q000502:physiology; D013879:Thioredoxins / Q000235:genetics / Q000502:physiology; D014471:Umbilical Veins / Q000166:cytology / Q000502:physiology
D002352:Carrier Proteins; D017127:Glycation End Products, Advanced; D015850:Interleukin-6; D016209:Interleukin-8; D016328:NF-kappa B; D000067759:Receptor for Advanced Glycation End Products; D011971:Receptors, Immunologic; C486715:TXNIP protein, human; D013879:Thioredoxins; D005947:Glucose