BACKGROUND: A series of studies was performed to examine the ability of estradiol (E2) to protect endothelial cells from apoptosis. METHODS AND RESULTS: Light and transmission electron microscopy demonstrated typical features of apoptosis in human umbilical vein endothelial cells (HUVEC) exposed to tumor necrosis factor-alpha (TNF-alpha). Northern and Western blot analyses revealed induction of message and protein for the interleukin-1 beta converting enzyme (ICE), which has been shown to mediate apoptosis induced by TNF-alpha. Immunofluorescent staining of HUVEC colocalized ICE expression to apoptotic HUVEC. Direct cell counting demonstrated a significant decrease in total endothelial cell number after 24 hours of TNF-alpha exposure and a dose-dependent reversal of the effect of TNF-alpha with E2 treatment. This protective effect was abrogated by an estrogen-receptor antagonist. Fluorescence-activated cell sorting analysis revealed 39.3% apoptosis after 24 hours of TNF-alpha exposure. Treatment with E2 resulted in a 50% decrease in apoptosis. Similarly, viability assays revealed 35 +/- 4% cell death after TNF-alpha exposure. Simultaneous treatment with E2 resulted in a dose-dependent reduction of cell death to a minimum of 18 +/- 2%. The protective effect of E2 was nullified by a specific estrogen-receptor antagonist. CONCLUSIONS: E2 treatment resulted in a dose-dependent, receptor-mediated inhibition of TNF-alpha-induced endothelial cell apoptosis. These studies indicate that E2 may also serve a maintenance function in preventing endothelial cell death after noxious stimuli and suggest that the ICE pathway may mediate cytokine-induced apoptosis in endothelial cells. Preservation of endothelial integrity represents another mechanism that may account for the atheroprotective effect of estrogen.
BACKGROUND: A series of studies was performed to examine the ability of estradiol (E2) to protect endothelial cells from apoptosis. METHODS AND RESULTS: Light and transmission electron microscopy demonstrated typical features of apoptosis in human umbilical vein endothelial cells (HUVEC) exposed to tumor necrosis factor-alpha (TNF-alpha). Northern and Western blot analyses revealed induction of message and protein for the interleukin-1 beta converting enzyme (ICE), which has been shown to mediate apoptosis induced by TNF-alpha. Immunofluorescent staining of HUVEC colocalized ICE expression to apoptotic HUVEC. Direct cell counting demonstrated a significant decrease in total endothelial cell number after 24 hours of TNF-alpha exposure and a dose-dependent reversal of the effect of TNF-alpha with E2 treatment. This protective effect was abrogated by an estrogen-receptor antagonist. Fluorescence-activated cell sorting analysis revealed 39.3% apoptosis after 24 hours of TNF-alpha exposure. Treatment with E2 resulted in a 50% decrease in apoptosis. Similarly, viability assays revealed 35 +/- 4% cell death after TNF-alpha exposure. Simultaneous treatment with E2 resulted in a dose-dependent reduction of cell death to a minimum of 18 +/- 2%. The protective effect of E2 was nullified by a specific estrogen-receptor antagonist. CONCLUSIONS:E2 treatment resulted in a dose-dependent, receptor-mediated inhibition of TNF-alpha-induced endothelial cell apoptosis. These studies indicate that E2 may also serve a maintenance function in preventing endothelial cell death after noxious stimuli and suggest that the ICE pathway may mediate cytokine-induced apoptosis in endothelial cells. Preservation of endothelial integrity represents another mechanism that may account for the atheroprotective effect of estrogen.
Authors: Maged M Costantine; Francesca Ferrari; Giusseppe Chiossi; Esther Tamayo; Gary D V Hankins; George R Saade; Monica Longo Journal: Am J Obstet Gynecol Date: 2009-07-26 Impact factor: 8.661