Journal of the Society for Gynecologic Investigation最新文献

Objective: The purpose of the current study was to determine whether maternal circulating components could regulate oxidative status of glutathione redox cycle and adhesion molecule expression in endothelial cells (ECs).

Methods: Maternal plasma was extracted from venous blood obtained from normal term pregnant women and from women with preeclampsia (PE). Normal and PE pregnancies were defined as American College of Obstetricians and Gynecologists criteria. Confluent ECs were incubated with EC growth medium (EGM) containing 20% plasma from women with normal (n = 8) and PE (n = 8) pregnancies for 4 hours. ECs incubated with EGM only were used as control. EC oxidative status was assessed by measuring cellular glutathione reductase (GR) and glutathione peroxidase (GPx) activities. Adhesion molecule expressions for intercellular adhesion molecule (ICAM), vascular cell adhesion molecule (VCAM), P-selectin, and E-selectin were determined by colorimetric assays detected on EC surface by UV spectrophotometer at OD 450 nm. Data are presented as mean +/- SE and analyzed by analysis of variance (ANOVA). A P value < .05 was set as statistically significant.

Results: Cellular GR activity was reduced approximately 35% in ECs treated with normal plasma and 70% in ECs treated with PE plasma compared to that in untreated control cells: 0.072 +/- 0.014 (P < .05), 0.039 +/- 0.006 (P < .01), versus 0.117 +/- 0.010 U/mg cellular protein, respectively. In contrast, GPx activity was slightly increased in ECs treated with normal plasma and significantly increased in ECs treated with PE plasma compared to that in untreated control cells: 0.059 +/- 0.005, 0.075 +/- 0.012 (P < .05) versus 0.044 +/- 0.002 U/mg cellular protein, respectively. P-selectin, E-selectin, and VCAM expressions were elevated in cells treated with normal plasma but significantly increased in cells treated with PE plasma compared to those of untreated controls: P-selectin--0.18 +/- 0.03, 0.35 +/- 0.04 versus 0.04 +/- 0.01 OD 450 nm, P < .01; E--selectin-0.06 +/- 0.02, 0.10 +/- 0.02 (P < .05) versus 0.03 +/- 0.01 OD 450 nm; VCAM--0.12 +/- 0.02, 0.16 +/- 0.03 (P < .01) versus 0.08 +/- 0.02 OD 450 nm, respectively. There was no difference for ICAM expression in cells treated with normal or PE plasma compared to controls.

Conclusions: These data suggest that endothelial pro- and anti-oxidative status could be directly affected by circulating components during pregnancy. Reduced cellular GR activity and increased GPx activity accompany increased inflammatory reactions in ECs responding to circulating "toxic" factors in preeclampsia.

Objective: The aim of the current study was to determine the effects of in vivo administration of prenatal betamethasone in patients at risk for preterm delivery on adrenomedullin (AM) concentrations in maternal and fetal plasma and on AM localization in placenta and fetal membranes.

Methods: A total of 62 pregnant women between 25 and 35 weeks' gestation were studied. Forty-seven pregnant women received betamethasone (2 x 12 mg intramuscularly given 24 hours apart) for stimulation of fetal lung maturity. Blood samples were collected before betamethasone administration and at different time points after the first and the second dose. Further samples were collected at delivery and, in women who did not deliver, after 1 week and 30 days from betamethasone administration. At delivery, placenta and membranes were collected. Fifteen patients who delivered at the same gestational age not receiving betamethasone represented the control group. AM concentration was determined by radioimmunoassay. Localization of AM in placental tissues was assessed by immunohistochemistry.

Results: Betamethasone caused approximately 50% increase in maternal plasma AM at 1 week after administration, whereas in fetal plasma AM levels increased by about 90% at 48 hours after betamethasone administration. There was increased immunohistochemical staining for AM in fetoplacental tissues collected after betamethasone administration.

Conclusion: These results provide the first evidence for in vivo stimulation of AM, likely of placental origin, by glucocorticoids in the third trimester human pregnancy.

Background: Miscarriage (early pregnancy failure) is a pregnancy-related disease, the pathophysiology of which is still not completely understood. Lipid peroxidation and alterations in antioxidant enzyme activities may be of importance in the pathogenesis of this disorder. This study was planned to investigate the possible relation between free radical scavenging enzyme activities and lipid peroxidation levels in placenta tissues with miscarriage.

Methods: Placental tissue samples were obtained from 21 patients who had miscarried and 25 normal pregnant women undergoing elective abortion as a control group. Total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) enzyme activities and levels of thiobarbituric acid reactive substances (TBARS), antioxidant potential (AOP), and nonenzymatic superoxide radical scavenger activity (NSSA) were measured in the placental tissues.

Results: GSH-Px, CAT activities, and TBARS levels were found to be significantly increased, while T-SOD and NSSA values decreased in patients with early pregnancy failure when compared with women undergoing elective abortion (control group). However, there were no significant differences in AOP levels between the groups.

Conclusions: Our results reflect oxidative stress in placenta tissues of early pregnancy failure, as the oxidative processes seem to be counteracted by the physiologic activation of antioxidant enzymes such as CAT and GSH-Px. Moreover, a compensatory mechanism might be developed against possible oxidative damage in patients with miscarriage.

Objective: The peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is a nuclear hormone receptor that plays an important role in many diseases. This study investigated whether two polymorphisms (Pro12Ala in exon B and C161T in exon 6) of the PPAR-gamma2 gene are related to adenomyosis, endometriosis, or leiomyomata.

Methods: A total of 390 patients with adenomyosis, endometriosis, and/or leiomyomata were classified into four groups: 103 patients with adenomyosis (21 adenomyosis only and 82 adenomyosis with endometriosis and/or leiomyomata), 95 patients with endometriosis only, 100 patients with leiomyomata only, and 92 patients with endometriosis and leiomyomata.

Results: There was no association between distribution of genotype or allele frequencies for the PPAR-gamma Pro12Ala polymorphism and the presence of adenomyosis, endometriosis, and/or leiomyomata. However, compared with results for controls, the PPAR-gamma 161CC genotype and 161C allele frequencies were significantly increased in patients with adenomyosis (genotype: chi2 = 8.185, corrected P value [Pc] = .0169; allele: chi2 = 8.337, Pc = .0155) and in patients with endometriosis (genotype: chi2 = 6.748, Pc = .0375; allele: chi2 = 6.413, Pc = .0453).

Conclusion: The results suggest that the PPAR-gamma 161CC genotype could be a genetic risk factor for adenomyosis and endometriosis, whereas the Pro12Ala polymorphism was not associated with these estrogen-dependent benign uterine diseases in a Japanese population.

Objective: Enhanced tyrosine phosphorylation of phospholipase C-gamma1 (PLCgamma1) is associated with increased spontaneous contractile activity. PLCgamma1 phosphorylation is regulated by cellular protein tyrosine kinases and tyrosine phosphatases (PTPs). The studies in this report were undertaken to characterize the expression of two PTPs known to bind to PLCgamma1: Src-homology phosphatase type-1 (SHP-1) and type-2 (SHP-2).

Methods: Uterine and other tissues were obtained from non-pregnant (estrus) and pregnant (gestational day 12 through day 1 postpartum) Sprague-Dawley rats. PTP activity in myometrial homogenates was determined using an in vitro fluorometric PTP assay with and without bpV(phen) (a nonselective PTP inhibitor), or PTP-Inhibitor 1 (PTP-I1, a SHP selective inhibitor). Western blots were performed using polyclonal antibodies to SHP-1 and SHP-2. Immunoprecipitation studies were performed to demonstrate an association between PLCgamma1 and the SHP proteins.

Results: The in vitro PTP assays demonstrated comparable enzyme activity in myometrium from estrus and pregnant animals. BpV(phen) produced a 93% reduction in PTP activity (P <.05); similarly, PTP-I1 produced an 86% reduction in enzyme activity (P <.05). Western blots confirmed robust expression of both SHP-1 and SHP-2 protein in rat uterus. SHP-1 expression decreased significantly at the end of gestation; in contrast, SHP-2 levels remained stable. Immunoprecipitation studies confirmed an association between the SHP proteins and PLCgamma1.

Conclusion: These studies have demonstrated that SHP-1 and SHP-2 are expressed in rat myometrium and appear to be responsible for the PTP activity in this tissue, thereby providing a molecular mechanism for the modulation of PLCgamma1 phosphotyrosine levels in the rat uterus.

Objective: The aim of the current study is to evaluate the effect of insulin-like growth factor-I (IGF-I) on the production of vascular endothelial growth factor (VEGF) in the human fallopian tube.

Methods: Human oviductal epithelial cells (OEC) and oviductal stromal fibroblasts (OSF) were isolated from the ampullary segment of the fallopian tubes of six premenopausal patients in the proliferative phase of the menstrual cycle. The secretion of VEGF165 by cultured OEC and OSF in response to IGF-I was measured using an enzyme-linked immunosorbent assay (ELISA).

Results: The secretion of VEGF165 was detected in cultured OEC and OSF under untreated conditions. The secretion of VEGF165 was significantly stimulated with IGF-I administration in these cells.

Conclusion: The present findings suggest that IGF-I in the local environment may stimulate oviductal vascular permeability by inducing the production of VEGF by oviductal cells through autocrine and paracrine mechanisms. The modulation of the VEGF production in the fallopian tube may contribute to the normal and pathologic processes of oviductal fluid secretion by regulating oviductal vascular permeability during the menstrual cycle and in the peri-implantation period.

Objective: Due to the extensive use of intracytoplasmic sperm injection (ICSI) in assisted reproduction, not only among couples with severe male factor infertility problems, but to a broader scale, a lot of concern has been raised regarding the safety of the method and its implications in epigenetic control and imprinting dysregulation. This review means to provide a comprehensive report of the published scientific data, outline putative associations between ICSI and epigenetic control, and suggest measures to improve the current state of affairs and reach more scientifically consolidated results.

Methods: This review was conducted by studying a broad spectrum of articles dealing with the subject of epigenetic control and its relation with ICSI. We tried to view the two subjects as parallel procedures that occur in the organism and by delineating the molecular and biochemical steps that comprise them make suggestions about putative associations between ICSI and epigenetic control.

Conclusions: No hard evidence presented at the moment can prove or disapprove ICSI's implications in epigenetic control. Nevertheless, we take the view that more comprehensive, long-term, and properly designed studies are imperative to be applied on a large-scale basis. We urge cautiousness, since the welfare of our progeny is what is at stake.

Objective: To test the hypothesis that fetal membranes (chorion or amnion) release one or more factors responsible for myometrial quiescence.

Methods: Myometrial samples were excised from women at elective term cesarean delivery prior to the onset of labor. Fetal membranes were obtained after cesarean delivery either before or during labor, and either term (greater than 37 weeks) or preterm (less than or equal to 36 weeks). Myometrial strips were placed in organ baths and contractions stimulated by oxytocin (10(-8) M). Contractility was measured under isometric conditions before and after exposure to fetal membranes or conditioned medium. The impact of either membrane or conditioned media on contractility was determined before and after myometrial K+ channel blockade.

Results: Both chorion and amnion and their respective conditioned mediums decrease oxytocin-stimulated myometrial contraction. The inhibitory effect was greatest with membranes from preterm pregnancies (mean gestation 32 weeks, P <.05). The inhibitory effect was detectable in the presence of term labor, but was absent when the fetal membranes were obtained after preterm labor. Iberiotoxin, an inhibitor of large conductance Ca2+-activated K+ channels (BK(Ca)) reduced the effect of fetal membranes by 50% (P <.05).

Conclusion: We conclude that human fetal membranes release one or more factors that inhibit oxytocin-induced myometrial contractility. We suggest this factor (or factors) acts mainly by opening myometrial BK(Ca). The findings further support our hypothesis that the fetal membranes release a factor (or factors) that is central to myometrial quiescence and its premature loss leads to preterm delivery.

Objectives: To compare lactate uptake in the microvillous plasma membrane (maternal facing [MVM]) in term and preterm placentas in intrauterine growth restriction (IUGR) and appropriate weight for gestational age (AGA) controls, and in the basal plasma membrane (fetal facing [BM]) at term. In addition, we examine the expression of monocarboxylate transporters (MCT1 and MCT4).

Methods: We measured [14C] L-lactate uptakes into vesicles prepared from MVM and BM, stimulated by an inwardly directed H+ gradient. MCT expression was examined by Western blotting.

Results: In term placentas, mean (+/- SE) [14C] L-lactate uptake into MVM vesicles of the IUGR (n = 6) and AGA (n = 11) groups at initial rate was similar (15.4 +/- 2.3 versus 15.0 +/- 1.1 pmol/mg protein/20 s). In preterm placentas, in IUGR (n = 3) and AGA (n = 3) groups, [14C] l-lactate uptake into MVM was also not significantly different. In BM vesicles from term placentas, [14C] L-lactate uptake was significantly lower in IUGR (n = 5) than in AGA (n = 6) controls (3.6 +/- 0.4 versus 5.6 +/- 0.6 pmol/mg protein/20 s, P <.05). MCT1 and MCT4 were expressed in BM vesicles, but there was no difference in expression between the IUGR and AGA groups.

Conclusions: These findings suggest that in IUGR placental lactate transport capacity in the BM is reduced, which may adversely affect placental lactate clearance.

Objective: To test if leflunomide, an immunomodulator, could impede the growth of an ectopic uterine tissue.

Methods: Endometriosis was surgically induced in 26 rats by transplanting an autologous fragment of endometrial tissue onto the inner surface of the abdominal wall. Four weeks later two rats were killed. The volume and weight of the implants were measured. The remaining rats were randomly grouped, and in group 1 no medication was given. To the rats in group 2, 35 mg/kg/d of leflunomide was administered orally. Four weeks later, rats were killed and ectopic uterine tissues were reevaluated morphologically and histologically. A scoring system was used to evaluate preservation of epithelia.

Results: Two rats in the control group died 5 weeks after surgery. There was a significant difference in post-treatment spherical volumes (139.1 +/- 92.8 versus 33.5 +/- 12.5 mm3) and explant weights (156.3 +/- 105.6 versus 38.6 +/- 12.6 mg) between the control and leflunomide-treated groups. The epithelia were found to be preserved significantly better in the control group when compared with the leflunomide-treated group (median 2.5 [interquartile range, 1.25] versus median 1.00 [interquartile range, 1.5]).

Conclusion: Leflunomide appeared to cause regression of experimental endometriosis in rats.