calcimycin and Pre-Eclampsia

To investigate whether decidual endothelial cells (DEC) contribute to the pathogenesis of preeclampsia through abnormal nitric oxide production. Decidual endothelial cells from normal (NDEC) and preeclamptic (PEDEC) pregnancies, and also human umbilical vein endothelial cells (HUVEC), were examined.. HUVEC, NDEC, and PEDEC were incubated for 45 min in serum-free media with the addition of potential stimulators [calcium ionophore (A23187), sepiapterin, and a combination of cytokines (TNF-alpha, gamma-IFN and LPS)], and the competitive inhibitor, NG-monomethyl-L-arginine (L-NMMA). These were added alone or in combination. Supernatants were measured for nitrate/nitrite (NOx) levels and the cells acid-extracted for measurement of cyclic guanosine monophosphate (cGMP). The effect of 30 min of shear stress (approximately 20 dynes/cm2) on NO and cGMP production by NDEC and PEDEC and on production of prostacyclin and thromboxane A2, was assessed.. PEDEC and HUVEC both produced more NO than NDEC under all conditions examined. Cell-associated cGMP levels, however, were not different among the cell groups but were increased by A23187 and inhibited by L-NMMA. In control conditions, shear stress stimulated cGMP levels 5-fold (p<0.01) in both NDEC and PEDEC, and PGI2 production 2-fold (p<0.05).. DEC from preeclamptic women do not have reduced NO production and respond normally to shear stress by increasing cGMP and PGI2 production. Our results are consistent with other reports of equal or higher NO levels in preeclampsia and indicate that reduced NO production by endothelial cells is not the explanation for the vasoconstriction of uterine vessels.

Topics: Adult; Calcimycin; Case-Control Studies; Cells, Cultured; Cyclic GMP; Cytokines; Decidua; Endothelium, Vascular; Epoprostenol; Female; Humans; Nitric Oxide; omega-N-Methylarginine; Pre-Eclampsia; Pregnancy; Pteridines; Pterins; Thromboxane A2; Umbilical Veins

Nitric oxide is an important vasodilator, and in this study we studied whether the calcium-dependent nitric oxide production capacity of human umbilical vein endothelial cells was affected by preeclampsia.. Human umbilical vein endothelial cells were isolated from 11 preeclamptic and 10 normotensive pregnancies. The maximal calcium ionophore A23187-stimulated nitric oxide production capacity was measured as accumulation of nitrate and nitrite into the culture medium, and it was related to the number of viable endothelial cells by measurement of their mitochondrial dehydrogenase activity.. The cell number-related nitric oxide production capacity was similar in preeclamptic and normotensive pregnancies. The total nitric oxide production of cells from preeclamptic pregnancies was significantly lower (p <0.001). This difference, however, was mainly caused by larger amount of viable endothelial cells recovered from normotensive pregnancies.. The maximal calcium-dependent nitric oxide production capacity of individual human umbilical vein endothelial cells is not affected by preeclampsia.

Topics: Adult; Analysis of Variance; Antihypertensive Agents; Calcimycin; Calcium; Cell Count; Cells, Cultured; Endothelium, Vascular; Female; Humans; Ionophores; Labetalol; Linear Models; Nitrates; Nitric Oxide; Nitrites; Pre-Eclampsia; Pregnancy; Umbilical Veins

The amount of Leukotriene B4 (LTB4) released from neutrophilic leukocyte activated by calcium ionophore was measured to examine neutrophilic functions regarding the changes during normal pregnancy and differences between normal pregnant women and preeclamptic women. In addition, we examined the arachidonic acid (AA) content and fatty acid composition of neutrophilic phospholipid in normal pregnant women. In a normal pregnancy, the amount of LTB4 released decreased significantly with gestation. In preeclamptic women, the amount of LTB4 released was significantly higher than that in normal pregnant women, but there was no significant difference from non-pregnant women. Normal pregnant women showed a significantly decreased AA content with gestation. Regarding other fatty acids, as to fatty acid composition, there was increased saturated fatty acid and decreased unsaturated fatty acid with gestation. The results suggested that the LTB4 released was inhibited more in pregnant women than in non-pregnant women, and pregnant women also had decreased AA content. In addition, changes in fatty acid compositions showed signs of decreased fluidity of the cell membrane. This phenomenon is thought to be due to a mechanism to inhibiting the activation of neutrophilic leukocytes accompanied by a decrease in the amount of LTB4 released. Conversely, no similar inhibition was observed in preeclamptic women, and the failure of this mechanism seemed to contribute to the development of preeclampsia.

Topics: Adult; Calcimycin; Cells, Cultured; Fatty Acids; Female; Humans; Ionophores; Leukotriene B4; Neutrophils; Pre-Eclampsia; Pregnancy

Endothelial cells isolated from umbilical veins (HUVEC) and from decidual biopsies collected at caesarean section delivery (DEC) from both normal (N DEC) and pre-eclamptic (PE DEC) women, were maintained in culture until passage 2, when the effect on growth of removing heparin/ECGS (endothelial cell growth supplement) from the culture medium was assessed, and the effects of heparin-free incubation and of the Ca2+ ionophore A23187 on endothelin-1, prostacyclin and prostaglandin E2 secretion over a 24 h period were examined. Cell growth slowed significantly in all three cell types in the absence of heparin/ECGS, and cell death occurred in 1/3 samples of HUVEC, 4/6 of N DEC, but 0/2 of PE DEC over 4 days. During the 24 h incubation for prostaglandin in medium without these growth factors, there was further cell death in N DEC. The addition of A23187 to this stress led to a reduction in cell number in both N DEC and HUVEC, and to a lesser extent in PE DEC. Prostaglandin and endothelin-1 levels were higher in the absence of heparin/ECGS in all cell types There was significant suppression of endothelin-1 secretion at 24 h incubation, and stimulation of prostaglandin secretion by A23187. Incubation without heparin/ECGS magnified the effect of A23187 on prostaglandin secretion, although the proportional change was similar if compared to controls without heparin/ECGS. Withdrawal of heparin/ECGS from the medium altered the balance of PGE2/PGI2 secretion by HUVEC, but not DEC. Endothelial cells require the presence of heparin/ECGS for optimum growth and viability, and N DEC are particularly dependent on these growth factors. PE DEC appear relatively 'hardy' in this regard. The addition of a further potentially toxic stimulus may result in cell death, and experiments to be conducted in limited medium must take this into account. There are both qualitative and quantitative differences in the effects of these stimuli on secretion of vasoactive substances, between decidual and umbilical vein endothelial cells.

Topics: Calcimycin; Calcium; Cell Death; Cell Division; Cells, Cultured; Culture Media; Decidua; Endothelin-1; Endothelium, Vascular; Female; Growth Substances; Heparin; Humans; Pre-Eclampsia; Pregnancy; Prostaglandins; Umbilical Veins

Pre-eclampsia is characterized by an increased vascular tone which might be related to an abnormal endothelial cell function. As representatives of the fetal circulation, we compared the nitric oxide (NO)-releasing capacity of human umbilical vessels from normal and pre-eclamptic pregnancies.. Normal and pre-eclamptic umbilical vessels were mounted in parallel in an organ chamber with three perfusion lines superfusing the same detector tissue (rubbed rat aortic ring). In this cascade system the capacity of the umbilical vessels to release NO was measured under basal conditions and after stimulation with histamine, bradykinin or calcium ionophore A23187.. Relaxations dependent on basal NO release were found to be significantly higher in pre-eclamptic vessels (especially in veins) than in normal vessels. Conversely, stimulated NO release in response to histamine or bradykinin was significantly decreased in pre-eclamptic umbilical arteries, but not in veins, compared with normal vessels. However, there was no significant difference in the release of NO in response to A23187 between normal and pre-eclamptic vessels.. The NO-releasing and NO-producing capacity in the vessels from fetal circulation is not diminished in pre-eclampsia. However, in pre-eclamptic umbilical arteries the NO release in response to certain stimuli (histamine or bradykinin) is diminished, probably as a result of alterations in the receptor function.

Topics: Adult; Animals; Aorta; Biological Assay; Calcimycin; Female; Histamine; Humans; In Vitro Techniques; Nitric Oxide; Norepinephrine; Pre-Eclampsia; Pregnancy; Rats; Umbilical Arteries; Umbilical Veins; Vasoconstriction; Vasodilation

To elucidate the relationship between the high concentration of taurine in platelets and platelet aggregation in patients with EPH gestosis (gestosis with edema, proteinuria and hypertension), platelet aggregation and the platelet release response (release of ATP and beta-thromboglobulin) were studied in the washed platelet suspension (PS) obtained from normal pregnant or non-pregnant women and EPH gestosis patients. Platelet aggregation and platelet release response were significantly lower in EPH gestosis patients than in normal pregnant and non-pregnant women. Platelet aggregation, platelet release response induced by ADP and collagen and the aggregation induced by A23187 were inhibited in taurine-loaded PS from non-pregnant women. These results suggest that the decrease of platelet aggregation in EPH gestosis patients was caused by high concentrations of taurine in platelets, which may inhibit the intracellular Ca2+ movement and platelet release response. Therefore, taurine appears to have a protective effect against the hyper-coagulative state in EPH gestosis.

Topics: Adenosine Diphosphate; Adenosine Triphosphate; beta-Thromboglobulin; Blood Platelets; Calcimycin; Collagen; Female; Humans; In Vitro Techniques; Platelet Aggregation; Platelet Aggregation Inhibitors; Pre-Eclampsia; Pregnancy; Taurine