Positive versus negative effects of VEGF165 on Ca2+ signaling and NO production in human endothelial cells.
The role increased vascular endothelial growth factor (VEGF) plays in vascular function during normal vs. preeclamptic pregnancy has been a source of some controversy of late. In this study, we seek to understand how VEGF165influences vasodilator production via Ca2+signaling mechanisms in human endothelial cells. We utilize human umbilical vein endothelial cells (HUVEC) as well as intact ex vivo human umbilical vein (HUV Endo) to address direct stimulation of Ca2+and NO by VEGF165alone, as well as the effect of VEGF165on subsequent ATP-stimulated Ca2+signaling and NO production. We show that VEGF165stimulates Ca2+responses in both HUVEC and HUV Endo, which results in a corresponding increase in NO production in HUV Endo. Longer-term VEGF165pretreatment then inhibits sustained Ca2+burst responses to ATP in HUVEC and HUV Endo. This is paralleled by a corresponding drop in ATP-stimulated NO production in HUV Endo, likely through inhibition of Cx43 gap-junction function. Thus, although VEGF165makes a small initial positive impact on vasodilator production via direct stimulation of Ca2+responses, this is outweighed by the greater subsequent negative impact on Ca2+bursts and vasodilator production promoted by more potent agonists such as ATP. Overall, elevated levels of VEGF165associated with preeclampsia could contribute to the endothelial dysfunction by preventing Ca2+bursts to other agonists including but not limited to ATP.
In this manuscript, we show that VEGF levels associated with preeclampsia are a net negative contributor to potential vasodilator production in both a human ex vivo and in vitro endothelial cell model. Therefore, pharmacological targeting of VEGF-stimulated signaling pathways could be a novel treatment modality for preeclampsia-related hypertension.
Perinatal Research Laboratories, Department of Obstetrics and Gynecology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin; and.
Derek S
DS
Boeldt
Perinatal Research Laboratories, Department of Obstetrics and Gynecology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin; and.|Division Maternal Fetal Medicine, Department of Obstetrics and Gynecology, School Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin.
Jennifer
J
Krupp
Perinatal Research Laboratories, Department of Obstetrics and Gynecology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin; and.
Fu-Xian
FX
Yi
Perinatal Research Laboratories, Department of Obstetrics and Gynecology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin; and.|Division Maternal Fetal Medicine, Department of Obstetrics and Gynecology, School Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin.
Nauman
N
Khurshid
Division Maternal Fetal Medicine, Department of Obstetrics and Gynecology, School Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin.
Dinesh M
DM
Shah
Perinatal Research Laboratories, Department of Obstetrics and Gynecology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, Wisconsin; and imbird@wisc.edu.
Ian M
IM
Bird
D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects*; D018031:Connexin 43 / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005260:Female; D017629:Gap Junctions / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D009569:Nitric Oxide / Q000096:biosynthesis*; D011225:Pre-Eclampsia / Q000378:metabolism*; D011247:Pregnancy; D014471:Umbilical Veins / Q000187:drug effects* / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology*
D018031:Connexin 43; C500216:GJA1 protein, human; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; D009569:Nitric Oxide; D002118:Calcium
Ca2+; endothelium; human; nitric oxide; pregnancy