Increased superoxide generation and decreased stress protein Hsp90 expression in human umbilical cord vein endothelial cells (HUVECs) from pregnancies complicated by preeclampsia.
OBJECTIVE
Endothelial dysfunction is associated with increased oxidative stress in the vascular system in women with preeclampsia (PE), a hypertensive disorder occurring during human pregnancy. However, due to the nature of the disease, direct evidence of increased endothelial oxidative stress in the maternal vascular system at an in vivo situation is still lacking. We previously reported that primary cultured endothelial cells (ECs) from umbilical cords (HUVECs) from pregnancies complicated by PE exhibit phenotypic changes compared to those from normal pregnancies such as reduced eNOs expression associated with disorganized endothelial junction protein distribution and increased endothelial permeability. In this study, we sought to determine whether increased oxidative stress was also present in primary cultured HUVECs from women with PE.


METHODS
HUVECs were isolated from normal and PE pregnancies and EC oxidative stress was examined by superoxide generation using positive nuclear dihydroethidium (DHE) staining as an indicator. Since Hsp90 is believed to have protective effects on endothelial function, we also determined mRNA and protein expression for Hsp90. Using Hsp90 inhibitor geldanamycin (GA), we further determined the potential role of Hsp90 in superoxide generation, eNOs expression, and prostacyclin production of altered EC function associated with PE pregnancies.


RESULTS
We found that primary cultured ECs from PE pregnancies showed an increase in DHE positive cells, p < 0.01. Hsp90 protein expression was significantly decreased in ECs from PE compared with that from normal pregnancies, p < 0.05. Inhibition of Hsp90 by GA resulted in an increase in superoxide generation and a decrease in eNOs protein expression. Decreased prostacyclin production was also found in ECs treated with GA.


CONCLUSIONS
These in vitro HUVEC data suggest that increased endothelial oxidative stress may also occur in the fetal compartment during preeclampsia.
Gu
Yang
Y
Department of Obstetrics and Gynecology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130, USA.
Lewis
David F
DF
Zhang
Yanping
Y
Groome
Lynn J
LJ
Wang
Yuping
Y
D016227:Benzoquinones; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism*; D011464:Epoprostenol / Q000378:metabolism; D005260:Female; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D047029:Lactams, Macrocyclic; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology; D011225:Pre-Eclampsia / Q000378:metabolism*; D011247:Pregnancy; D013481:Superoxides / Q000378:metabolism*; D014471:Umbilical Veins / Q000378:metabolism*
D016227:Benzoquinones; D018841:HSP90 Heat-Shock Proteins; D047029:Lactams, Macrocyclic; D013481:Superoxides; D011464:Epoprostenol; D052250:Nitric Oxide Synthase Type III; C001277:geldanamycin