ELD increased PPARγ expression and decreased p65 expression in OVX rat femoral arteries. ELD also decreased Nox4 expression and nitrotyrosine content.
Endothelial function and vascular remodelling
sis but also endothelial dysfunction, leading to cardiovascular events. Eldecalcitol (ED-71; ELD), a 2β-hydroxypropyloxy derivative of 1α,25 (OH)2D3, is a novel active vitamin D3 analog for the treatment of osteoporosis. The aim of this study was to evaluate the endothelial protective effect of ELD in ovariectomized (OVX) rats. Methods: Female Sprague-Dawley rats (12 weeks) were ovariectomized. ELD (20 ng/kg, the effective dose for improvement of bone mineral density) was orally administrated 5 times a week for 4 weeks from 1 day after surgery. Four weeks after surgery, flow-mediated dilation (FMD) as an indicator of endothelial function was measured by high-resolution ultrasound in the femoral artery of anesthetized rats. After that, the femoral artery was harvested and used for western blot analysis. Results: FMD was significantly decreased in OVX rats as compared with sham-operated rats. ELD reversed the reduction of FMD in OVX rats (Sham, 14.3±2.5%; OVX, 8.2±1.5%; OVX+ELD, 15.0±1.1%; mean ± SE, n=7–10). In OVX rats, PPARγ expression was decreased in femoral arteries, leading to an increase in NF-κB p65 expression. Furthermore, Nox4 expression (a NADPH oxidase component) and nitrotyrosine content (a maker of oxidative stress such as peroxynitrite) were also increased in OVX rat femoral arteries. Increased oxidative stress led to a decrease in dimer/monomer ratio of eNOS in OVX rat femoral arteries. ELD increased PPARγ expression and decreased p65 expression in OVX rat femoral arteries. ELD also decreased Nox4 expression and nitrotyrosine content in OVX rat femoral arteries and improved eNOS uncoupling state. Conclusion: These results suggest that ELD ameliorated endothelial dysfunction in OVX rats through improvement of eNOS uncoupling state by an antioxidative effect. This antioxidative effect may be mediated by normalization of the PPARγ/NF-κB signaling pathway. ELD might improve the prognosis of postmenopausal women by improving not only bone mineral density but also endothelial function.
P592 | BENCH Isolation and characterisation of endothelial outgrowth from coronary arteries in patients with acute myocardial infarction M. Brittan, S. Gallogly, N.L. Mills. University of Edinburgh, Centre for Cardiovascular Science, Edinburgh, United Kingdom Purpose: Our understanding of endothelial cell biology is primarily derived from studies of human umbilical vein endothelial cells (HUVECs). However, HUVECs provide limited insight into disease pathogenesis. We describe a novel method for isolation of coronary artery endothelial cells from thrombectomy specimens obtained during the treatment of patients with acute myocardial infarction. We have expanded and characterised human coronary endothelial outgrowth (CEO) cells compared to HUVECs, to investigate their potential as a model of endothelial dysfunction. Methods: Patients presenting to the Edinburgh Royal Infirmary with ST-segment elevation myocardial infarction (n=15) underwent emergency percutaneous coronary intervention and thrombus aspiration. Thrombus specimens were dissected, plated onto collagen-I coated plates and maintained in vitro to encourage cellular outgrowth. Population doubling time was evaluated and angiogenic potential assessed by tubule formation. Multiparameter flow cytometry using antibodies to endothelial cell markers was performed (CD31, KDR, CD146 and CD34) and cells were immunostained for vonWillenbrand factor (vWF). Results: CEO cell outgrowth was observed in 9/15 samples. Population doubling times of CEO cells were comparable to HUVECs (mean±SD: CEO 2.6±0.5, HUVEC 2.3±0.1 days; p=0.50 student’s t-test). CEO cells had typical "cobblestone" morphology and were immunoreactive for vWF. Surface expression of CD31 and KDR was comparable in both cell types (CD31 mean±SD: CEO 74.9±21.3 vs. HUVEC 89.3±11.4, p=0.13; KDR mean±SD: CEO 46.7±31.7 vs. HUVEC 22.2±15.9, p=0.11), however CD146 and CD34 expression was increased in CEO cells (CD146 mean±SD: CEO 96±5.3 vs. HUVEC 73.5±30.7, p
