Biochemical and Biophysical Research Communications
TNF-α upregulates the A2B adenosine receptor gene: The role of NAD(P)H oxidase 4
Section snippets
Materials and methods
Vascular smooth muscle cell culture conditions. Aortic VSMC were isolated from neonatal Sprague–Dawley rats, as described previously [26], [27], and were seeded at a density of 1.5 × 105 cells per well of a 6-well plate; all experiments were performed with cells in their first passage. VSMC were cultured in VSMC growth medium containing DMEM, 10% bovine calf serum (BCS), 1 mM sodium pyruvate, 1 mM nonessential amino acids, 100 U/mL penicillin, and 100 μg/mL streptomycin (all from Invitrogen,
Effect of an inflammation-inducing agent and TNF-α on A2BAR mRNA and promoter activity in vitro and in vivo
In addition to our findings describing the inducibility of the A2BAR gene in proliferating VSMC by the transcription factor B-Myb [30], a recent paper by Kong et al. identified HIF-1α as the responsible transcription factor for the induction of A2BAR expression in endothelial cells under hypoxic conditions [31]. In the context of atherosclerosis, proliferation and hypoxia are stress responses, and A2BAR activation has been shown to be protective in these instances [14], [31]. Additionally,
Acknowledgments
We thank Dr. Barbara Schreiber and the cell core for preparation of smooth muscle cells and for the insight, as well as Dr. Xu Yong for initial assistance with qPCR. This work was supported by National Institutes of Health (NIH) Public Health Services Grant HL13262. K.R. is an Established Investigator with the American Heart Association.
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Present address: Laboratory for Comparative Carcinogenesis, National Cancer Institute-Frederick, Building 538, Room 144, Frederick, MD 21702, USA.