RESUMEN
OBJECTIVES: To test the hypothesis that the common missense mutation of 5,10-methylenetetrahydrofolate reductase (MTHFR) (677 C to T, ala to val) is more prevalent among nulliparous preeclamptic women compared with control and transient hypertension of pregnancy patients. The correlation of the MTHFR T677/T677 genotype in mothers and fetuses was also investigated to test for possible maternal-fetal interactions. Lastly, possible differences in serum folate concentrations between control and preeclampsia patients and the possibility of a correlation between serum folate and MTHFR genotype were investigated as well. METHODS: The MTHFR genotype was determined for 114 control subjects, 99 preeclamptic patients, and 24 patients with transient hypertension of pregnancy by a polymerase chain reaction/restriction fragment length polymorphism (PCR) method. To ensure homogeneity of ethnic background, only samples from white women were analyzed. Results were analyzed with a chi 2 test for homogeneity. Serum folate was determined by radioimmunoassay (RIA). RESULTS: The prevalence of the MTHFR T677/T677 genotype was not significantly different between the populations studied. There was no significant difference in the prevalence of the MTHFR T677/T677 genotype between the infants of preeclamptic and control mothers. Furthermore, there was no difference in serum folate concentrations between control and preeclampsia patients, and there was no correlation between serum folate and MTHFR genotype. CONCLUSION: These data suggest that contrary to previous published reports, the C677T missense mutation of MTHFR is not a risk factor for preeclampsia in this nulliparous patient population. Furthermore, this mutation is not related to serum folate status in late pregnancy.
Asunto(s)
Ácido Fólico/sangre , Predisposición Genética a la Enfermedad , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/genética , Polimorfismo de Longitud del Fragmento de Restricción , Preeclampsia/enzimología , ADN/sangre , Femenino , Sangre Fetal/enzimología , Genotipo , Humanos , Metilenotetrahidrofolato Reductasa (NADPH2) , Reacción en Cadena de la Polimerasa , Preeclampsia/genética , EmbarazoRESUMEN
The pathogenesis of preeclampsia is proposed to be due to uncharacterized circulating factors that activate endothelial cells. Support for this hypothesis is provided by in vitro activation of endothelial cells by plasma from preeclamptic women, eg, increased nitric oxide and prostacyclin generation. We performed molecular sizing, lipid extraction, and lipoprotein fractionation of plasma from normal pregnant and preeclamptic women and determined the ability of these plasma fractions to increase nitric oxide or prostacyclin generation by endothelial cells. Fractions from plasma of preeclamptic women were consistently more active than fractions from normal pregnant women, although characterization was qualitatively similar. The factors stimulating nitric oxide and prostacyclin were different. The factor (or factors) stimulating nitric oxide generation was extractable by charcoal and present in lipid extracts and lipoprotein isolates with a molecular weight greater that 1.5 million daltons, which is characteristic of a lipoprotein or lipoprotein aggregate. By contrast, activity to stimulate prostacyclin persisted after charcoal stripping or lipoprotein removal, partitioned to the aqueous fraction, and had a molecular weight of approximately 50,000 D. Two distinct factors in the blood of preeclamptic women alter endothelial function in vitro. This information should guide the search for circulating factors contributing to the pathophysiology of preeclampsia.
Asunto(s)
6-Cetoprostaglandina F1 alfa/sangre , Endotelio Vascular/metabolismo , Óxido Nítrico/biosíntesis , Óxido Nítrico/sangre , Preeclampsia/sangre , Células Cultivadas , Epoprostenol/biosíntesis , Femenino , Humanos , Lipoproteínas/aislamiento & purificación , Lipoproteínas/metabolismo , Peso Molecular , EmbarazoRESUMEN
OBJECTIVE: In preeclampsia markers of endothelial activation (e.g., increased cellular fibronectin and activities that alter in vitro endothelial function (e.g., stimulation of nitric oxide and prostacyclin generation) are increased in the maternal circulation. We tested preeclamptic infant blood for these markers and activities and correlated these findings with fetal growth. STUDY DESIGN: Plasma was obtained from 17 term nulliparcus preeclamptic and normal pregnant women and their infants and from 8 additional preeclamptic mother-baby pairs from earlier gestations. Plasma cellular fibronectin and production of nitric oxide and prostacyclin by cultured endothelial cells exposed to 2% plasma were measured. RESULTS: Cellular fibronectin was higher in maternal plasma of preeclamptic than nonpregnant women (6.1 +/- 0.29 vs 4.2 +/- 0.27 microgram/ml, p < 0.01), as were stimulated endothelial nitric oxide and prostacyclin production (nitric oxide 42.5 +/- 3.9 vs 26.9 +/- 2.3 nmol nitrite/microgram protein/24 hours, p < 0.05; prostacyclin 261.7 +/- 31.2 vs 151.9 +/- 18.7 pg prostaglandin F1 alpha/microgram protein/24 hours, p < 0.05). In the preeclamptic infants cellular fibronectin was also greater (3.3 +/- 0.15 vs 2.6 +/- 0.14 microgram/ml, p < 0.01), as was endothelial nitric oxide production in response to the plasma (24.4 +/- 1.1 vs 21.4 +/- 0.09 mumol/L nmol nitrite/microgram protein/24 hours, p < 0.05). Prostacyclin production was not significantly different. In preeclamptic infants across a wide gestational age there was no correlation of endothelial activation and fetal growth. CONCLUSIONS: Infants of women with preeclampsia may be affected by endothelial dysfunction, as well as reduced uteroplacental perfusion.
Asunto(s)
Endotelio Vascular/fisiología , Sangre Fetal/fisiología , Preeclampsia/fisiopatología , Adulto , Células Cultivadas , Desarrollo Embrionario y Fetal , Epoprostenol/biosíntesis , Femenino , Fibronectinas/sangre , Humanos , Recién Nacido , Óxido Nítrico/biosíntesis , EmbarazoRESUMEN
Because of the potent mitogenic and vasoactive properties of endothelin-1 (ET-1) and the presence of its receptor in third trimester placenta, we postulated that ET-1 might be involved in human placental growth and vascularization during development. As an initial approach to test this hypothesis, placental ET receptors were characterized and quantified in each trimester of pregnancy. Membrane-rich particulates were prepared from first-, second-, and third-trimester villous human placenta obtained immediately after pregnancy termination or delivery. ET receptors were characterized by radioligand saturation analysis, ligand competition, and reverse transcription-polymerase chain reaction (RT-PCR) to determine the concentration, affinity, and specificity of ET binding sites, and to document the presence of specific ET-receptor subtype mRNA transcripts in placentas from each trimester. Kinetic determinations of 125I-labeled ET-1 binding yielded a Kd = 61 pM, consistent with the equilibrium determinations of 34 +/- 6 pM (n = 11). The concentration of ET receptors decreased significantly from 682 +/- 94 fmol/mg protein (n = 4) in the first trimester to 266 +/- 89 fmol/mg protein (n = 4) in the third trimester. Competition studies with unlabeled ET-1 indicated a single class of binding sites with a Ki = 49 +/- 5 pM (n = 9), whereas competition with ET-3 demonstrated binding sites with two affinities. The predominant sites had a Ki = 84 +/- 14 pM, similar to that for ET-1. The RT-PCR data confirmed that both ETA and ETB receptors mRNA transcripts are expressed in human placenta.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Placenta/química , Receptores de Endotelina/análisis , Secuencia de Bases , Membrana Celular/química , Membrana Celular/fisiología , Membrana Celular/ultraestructura , Femenino , Humanos , Datos de Secuencia Molecular , Placenta/fisiología , Placenta/ultraestructura , Reacción en Cadena de la Polimerasa , Embarazo , Primer Trimestre del Embarazo , Segundo Trimestre del Embarazo , Tercer Trimestre del Embarazo , ARN Mensajero/análisis , ARN Mensajero/genética , Receptores de Endotelina/genética , Receptores de Endotelina/fisiología , Transcripción GenéticaRESUMEN
Growth-retarded infants have a reduced risk of pulmonary morbidity. We used a naturally occurring model of growth retardation in rabbits to test the hypothesis that reduced risk might be related to precocious maturation of the alveolar beta-adrenergic response system in runted neonates. We confirmed that the weights of the fetuses were significantly different, depending on uterine position, as predicted by this model. However, we found no evidence of either increased beta-adrenergic receptor concentration or cyclic adenosine monophosphate generation in the smaller fetuses. These results indicate that reduced fetal size in this model of growth retardation does not result in accelerated maturation of alveolar beta-adrenergic responses in neonates.