Tobacco smoke dysregulates endothelial vasoregulatory transcripts in vivo.
We hypothesized that human smoking and its deleterious effects on endothelial function can be modeled by exposure of mice to tobacco smoke, and further that these changes would be reflected in gene regulation in vascular endothelium. We used for these studies a mouse strain that expresses green fluorescent protein under the control of an endothelial-specific promoter, Tie-2. Mice were exposed to sidestream smoke from reference cigarettes at 34 mg total suspended particulates/m3. After exposure for 5 days/wk for 1 and 6 wk, aortas were pooled from treatment and control groups. Endothelial cells were rapidly isolated by collagenase treatment followed by fluorescent activated cell sorting to yield populations of >95% purity. RNA isolated from >500 endothelial cells was amplified and analyzed on deeply representative long oligo microarrays. Transcripts dysregulated by >2.5-fold were confirmed by real-time PCR and selected proteins by immunofluorescent localization. In the endothelial cells, the observed more than threefold upregulation of complement factor H (Cfh), calcitonin receptor-like (Calcr1), and soluble epoxide hydrolase (Epxh2) may play a role in hypertensive responses of the vasculature to smoking. We have identified gene regulation in vivo in vascular endothelium that potentially underlies hypertensive responses to tobacco smoke.
Maresh
J Gregory
JG
Department of Internal Medicine-Cardiology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA
Xu
Huaxia
H
Jiang
Nan
N
Gairola
C Gary
CG
Shohet
Ralph V
RV
D000818:Animals; D001011:Aorta; D001483:Base Sequence; D017931:DNA Primers; D004730:Endothelium, Vascular / Q000503:physiopathology*; D004781:Environmental Exposure; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D009131:Muscle, Smooth, Vascular; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D042787:Receptor, TIE-2 / Q000235:genetics; D012906:Smoke / Q000009:adverse effects; D012907:Smoking / Q000009:adverse effects*; D014158:Transcription, Genetic
D017931:DNA Primers; D012906:Smoke; D049452:Green Fluorescent Proteins; D042787:Receptor, TIE-2