Improvement of Function and Morphology of Tumor Necrosis Factor-.ALPHA. Treated Endothelial Cells With 17-.BETA. Estradiol A Preliminary Study for a Feasible Simple Model for Atherosclerosis

Abstract

n normal vasculature endothelial cells (EC), a small amount of NO synthesized by endothelial nitric oxide synthase (eNOS) maintains normal vascular physiological function controls. 1,2 Endothelial cell dysfunction in eNOS expression by tumor necrosis factor-(TNF-) is involved in the pathogenesis of vascular diseases, for example atherosclerosis. 3 The related pathologic changes are increased expression of adhesion molecules, including intercellular adhesion molecule (ICAM)-1, vascular adhesion molecule (VCAM)-1, and E-selectin that enhances inflammatory cell infiltration in the vascular wall, platelet aggregation and adhesion on EC surfaces, and smooth muscle cell proliferation and migration. 1,4 In contrast, 17-estradiol (E2) upregulates eNOS production in EC, protects against atherosclerosis and recovers EC after arterial injury in animal models. [5][6][7] At present, experimental models for pathological and pharmacological studies in atherosclerosis have been developed only in laboratory models, eg, primates, rats and mice. 5,6,8 The important defects in the animal models are their complicated metabolisms with systemic variation and the difficulty of controlling the concentrations of drugs acting directly on the target cells. To date, no in vitro vascular model has been demonstrated for pharmacological studies in atherosclerosis, even though there have been several studies with other vascular diseases. [9][10][11] In the present study, we first constructed the in vitro model vascular wall with endothelial cell dysfunction in eNOS production by TNF-to mimic the important basic early stage pathology of atherosclerosis, and determined the potential of the model by using E2 as an antiatheroslerogenic agent. Methods Endothelial Cell CultureHuman umbilical vein endothelial cells (HUVEC) were separated from fresh umbilical cord veins, as previously described. 12 The HUVEC were cultured in human endothelial-SFM basal growth medium (Gibco BRL, Gaithersburg, MD, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS), 40 g/ml gentamycin and glutamine. The HUVEC were incubated at 37°C in a 5% CO2 atmosphere. Positive staining for CD31 confirmed the endothe- Background Dysfunction of endothelial cells (EC) to produce endothelial nitric oxide synthase (eNOS) by tumor necrosis factor-(TNF-) causes critical features of vascular inflammation associated with several disease states (eg, atherosclerosis including increased platelet aggregation and adhesion on EC, elevated adhesion molecules and enhanced inflammatory cells binding to EC). 17-estradiol (E2) can stimulate eNOS production and improve the critical features of atherosclerosis. Using TNF-and E2, we attempted to develop an in vitro vascular model for studying atherosclerosis. Methods and ResultsHuman umbilical vein endothelial cells (HUVEC) grown in transwells were cocultured with smooth muscle cells in a 24-well plate to mimic the major components of the vascular wall. The model was incubated with TNF-(10 ng/ml) for 12 h, prior exposed to E2 (100 pg/ml) f...

Pregnancy-Related Hyperlipidemia and Endothelial Function in Healthy Women

“…18 E2 protects the endothelium by upregulating endothelial nitric oxide synthase and can reverse endothelial dysfunction in an in vitro vascular model. 19 In the present study we did not measure estrogen or other hormonal levels so we could not examine the correlation between E2 and FMD. In pregnancy the naturally occurring increase in E2 seems to protect the endothelium against the adverse lipid changes.…”

Delivery May Affect Arterial Elasticity in Women

“…15 Cevic et al demonstrated that a high concentration of estrogen reduces the level of asymmetric dimethylarginine (ADMA), which is an endogenous competitive inhibitor of nitric oxide (NO) synthase. 16 Estradiol, by reducing ADMA, may therefore facilitate NO synthesis in endothelial cells.…”

Mechanisms Underlying Nano-Sized Air-Pollution-Mediated Progression of Atherosclerosis Carbon Black Causes Cytotoxic Injury/Inflammation and Inhibits Cell Growth in Vascular Endothelial Cells

“…25 ECs apparently have to physically uncouple in order to allow transendothelial migration of leukocytes. Inflammatory mediators, such as tumor necrosis factor- 12,26 and lipopolysaccharide, 27 induce leukocyte adhesion molecules such as VCAM-1, but also downregulate connexin37 expression in vascular ECs. 28,29 The inhibition of connexin37 expression by CB thus seems to be associated with EC inflammatory process such as adhesion and/or transmigration of leukocytes.…”