Title |
Differential expression and role of hyperglycemia induced oxidative stress in epigenetic regulation of β1, β2 and β3-adrenergic receptors in retinal endothelial cells
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Published in |
BMC Medical Genomics, May 2014
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DOI | 10.1186/1755-8794-7-29 |
Pubmed ID | |
Authors |
Sher Zaman Safi, Rajes Qvist, Gracie Ong Siok Yan, Ikram Shah Bin Ismail |
Abstract |
Aberrant epigenetic profiles are concomitant with a spectrum of developmental defects and diseases. Role of methylation is an increasingly accepted factor in the pathophysiology of diabetes and its associated complications. This study aims to examine the correlation between oxidative stress and methylation of beta1, beta2 and beta3-adrenergic receptors and to analyze the differential variability in the expression of these genes under hyperglycemic conditions METHODS: Human retinal endothelial cells were cultured in CSC complete medium in normal (5 mM) or high (25 mM) glucose to mimic a diabetic condition. Reverse transcription PCR and Western Blotting were performed to examine the expression of beta1, beta2 and beta3-adrenergic receptors. For detections, immunocytochemistry was used. Bisulfite sequencing method was used for promoter methylation analysis. Apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay. Dichlorodihydrofluorescein diacetate (DCFH-DA) assay was used to measure reactive oxygen species (ROS) production in the cells. |
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