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PubMed1998

Apoptosis and Fas expression in human fetal membranes.

Apoptosis (i.e. programmed cell death) plays a key role in maintaining reproductive function in the ovary, mammary and prostate glands, uterus, and testis. The purpose of the present report was to determine, based on biochemical and morphological parameters, whether cells in human fetal membranes undergo apoptosis and express Fas (CD95), a cell surface receptor that mediates apoptosis. Using the terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling immunohistochemical technique, apoptotic nuclei were identified in amnion epithelial, chorionic trophoblast, and decidua parietalis cell layers of human fetal membranes at term. Electron microscopy of fetal membranes revealed ultrastructural characteristics in amnion epithelium and chorion trophoblast cell layers consistent with apoptosis, including condensation of chromatin along the periphery of the nucleus and nuclear shrinkage. The apoptotic index (percentage of terminal deoxynucleotidyl transferase deoxy-UTP-nick end labeling-positive nuclei of the total nuclei) ranged from 8-29% in amnion epithelial, chorionic trophoblast, and decidual cell layers from women at 23-30, 31-36, and 37-42 weeks gestation. The apoptotic index was statistically greater in the 37-42 week group than in the 23-30 week group in chorionic trophoblast (P < 0.05) and decidual cell (P < 0.01) layers. In contrast, the apoptotic index in the amnion epithelial cell layer was statistically greater (P < 0.05) in the 23-30 week group than in the 31-36 week group, suggesting that apoptosis may be independently regulated in amnion epithelial, chorionic trophoblast, and decidual cell types. Based on the importance of Fas in mediating apoptosis, we investigated whether Fas was expressed by human fetal membrane cells. Immunohistochemical staining of fetal membranes with anti-Fas antibody localized Fas in amnion epithelial, chorionic trophoblast, and decidua parietalis cell layers. A 266-bp band corresponding to the cytoplasmic domain of Fas was detected in samples of amnion, chorion, decidua, and placenta by RT-PCR. Northern blotting revealed a molecular weight of approximately 1.9 kilobases for Fas messenger ribonucleic acid in amniotic tissue. These data suggest that apoptosis and Fas signaling may play a role in remodeling of fetal membrane architecture across gestation.

Amnion↗
PubMed1998

Interrelationships between oxytocin and eicosanoids in human fetal membranes at term gestation: which role for leukotriene B4?

The existence of a functional paracrine loop between oxytocin and prostaglandin F2-alpha in human placental cells has been demonstrated. The present study was undertaken to investigate further the possible interrelationships between oxytocin and eicosanoids in human intrauterine tissues at term gestation. Therefore, we evaluated the effect of leukotriene B4 (LTB4) on oxytocin (OT) production by explants of fetal membranes and amnion and the effect of oxytocin on the production of LTB4 and prostaglandin E2 (PGE2) by both fetal membranes and amnion. In all cases studied (n = 25), short-term cultures of tissue explants (fetal membranes or amnion) have been carried out. The production of eicosanoids and oxytocin in culture medium was evaluated. Oxytocin measurement was carried out by radioimmunoassay following extraction of the substance with Sep Pak C18 cartridges, PGE2 and LTB4 were measured by radioimmunoassay directly in culture medium. Results show that LTB4 has no significant stimulatory effect on oxytocin production by fetal membranes or amnion tissue. On the other hand, oxytocin stimulates PGE2 release by both fetal membranes and isolated amnion, but has no effect on LTB4 production by these tissues. Taken together, these findings suggest the following conclusions: (1) a paracrine loop between LTB4 and oxytocin is lacking in fetal membranes and amnion at term pregnancy; (2) oxytocin exerts a stimulatory effect on PGE2 release by both fetal membranes and amnion; (3) the interrelationships between oxytocin and the different eicosanoids in the above tissues seem to be highly selective.

Adult↗
PubMedDate not supplied

The morphometry of the Sudan-III-positive granules in the cytoplasm of the human amniotic epithelium.

Seventy-nine human amnions from different gestational ages were studied by new morphometric methods. The Sudan-III-positive granules in the cytoplasm of the amniotic epithelial cells were first observed in the reflected amnion from the second trimester of normal gestation. During the third trimester of normal gestation, the number of cytoplasmic granules increased remarkably with gestational age not only in the reflected amnion but also in the placental amnion. The number of granules was generally related to the increasing rates of anucleated cells and binucleated cells in the amniotic epithelium during the third trimester of normal gestation. During the third trimester of gestation, the number of granules was greater in the distant reflected amnion with its less adequate blood supply than in the placental amnion. A vast accumulation of the granules in the entire amnion was observed in cases of fetal death, in which blood supply had ceased. It is suggested, therefore, that the progressive accumulation of Sudan-III-positive cytoplasmic granules in the human amniotic epithelium during the third trimester of normal gestation is the result of cell degeneration.

Amnion↗
PubMedDate not supplied

Expression of interleukin-10 in human gestational tissues.

OBJECTIVE: To determine production of interleukin-10 (IL-10) by interleukin-1 beta (IL-1 beta) in cultured decidual, chorion, and amnion cells and whether IL-10 is produced in gestational tissues under the setting of infection-associated preterm labor. METHODS: Decidual, chorion, and amnion cells were isolated from term placentas and grown in primary culture. The cells were incubated with various concentrations of IL-1 beta and then culture supernatants were assayed for IL-10 by enzyme-linked immunosorbent assay. In subsequent studies, gestational membranes were isolated from a normal-term pregnancy and a preterm pregnancy complicated by chorioamnionitis. Tissues were evaluated for IL-10 expression by immunohistology and in situ hybridization. Human gestational tissues were collected from 38 women experiencing: 1) term cesarean delivery without labor; 2) normal-term vaginal delivery; 3) preterm cesarean delivery without labor; 4) preterm vaginal delivery without chorioamnionitis; and 5) preterm vaginal delivery with concomitant chorioamnionitis. Amnion, chorion, and decidua were isolated, total RNA from each tissue was extracted, and the presence of IL-10 mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: Decidual cells in culture produced IL-10 in response to IL-1 beta, but chorion and amnion cells produced no IL-10 protein. In vivo protein expression by immunohistology showed that most protein was detected within decidua while cells within amnion and chorion rarely had detectable IL-10 protein. In vivo RT-PCR samples demonstrated the strongest IL-10 mRNA signal from decidua samples, although IL-10 mRNA was also noted in chorion and amnion of placentas obtained after preterm labor. CONCLUSION: Maternal decidual cells can potentially produce IL-10, but fetal membranes (amnion and chorion) appear to have limited capabilities to produce IL-10. The relative inability of fetal tissues to produce IL-10 may play an important role in the pathophysiology of infection-associated preterm labor.

Amnion↗
PubMed1990

Mechanisms of parturition: the transfer of prostaglandin E2 and 5-hydroxyeicosatetraenoic acid across fetal membranes.

Prostaglandin E2 production by amnion is thought to be an important event in the onset of human labor. It has been suggested that 5-hydroxyeicosatetraenoic acid, also produced in the amnion, may mediate prelabor contractions. For either of these compounds to play a paracrine role they need to cross the chorion, which has a high capacity to metabolize prostaglandins. With the use of an in vitro system we have shown that both prostaglandin E2 and 5-hydroxyeicosatetraenoic acid cross either amnion or intact amnion-chorion-decidua at a rate similar to that of an extracellular marker sucrose. Analysis by high performance liquid chromatography revealed that at physiologic concentrations neither prostaglandin E2 nor 5-hydroxyeicosatetraenoic acid were metabolized by amnion alone. Moreover, 100% of the 5-hydroxyeicosatetraenoic acid and 72% of the prostaglandin E2 remained in the active form after passage across intact amnion-chorion-decidua. There did not appear to be any difference in the rate of transfer or permeability of the membranes before or after spontaneous labor. We conclude that both 5-hydroxyeicosatetraenoic acid and prostaglandin E2 synthesised in the amnion can cross the membranes by diffusion through the extracellular space, remaining largely unmetabolized, and may play a role in the onset of human labor.

Biological Transport↗
PubMed2006

Labour induces increased concentrations of biglycan and hyaluronan in human fetal membranes.

OBJECTIVE: The proteoglycan decorin stabilizes collagen whereas biglycan and hyaluronan disrupt well-organized collagen. The aim was to compare hyaluronan and proteoglycans in human fetal membranes obtained before and after spontaneous labour at term. STUDY DESIGN: Prelabour samples of fetal membranes (N=9) were obtained from elective caesarean sections and regionally sampled from over the cervix (cervical membranes) and mid-zone samples between this area and the placental edge. Postlabour samples (N=11) were obtained from spontaneous vaginal delivery and also regionally sampled. Amnion and chorio-decidua were analysed separately. The proteoglycans decorin and biglycan were analysed using alcian blue precipitation, SDS polyacrylamide gel electrophoresis and immunostaining. Hyaluronan was analysed using a radioimmunoassay and by histochemistry. Collagen was measured by estimating hydroxyproline content. RESULTS: In prelabour membranes the biglycan concentration (microg/mg wtw) in the cervical amnion was 40% lower than in the mid-zone amnion (P<0.05). After delivery the cervical amnion showed a twofold increase in biglycan (P<0.05), a 30% decrease in collagen (P<0.05), and a 50% decrease in decorin concentration (P<0.05). In mid-zone samples after delivery the concentrations of hyaluronan showed an increase form 1.0 to 4.9 microg/mg wtw (P<0.05). Histology demonstrated a gelatinous substance, which separated amnion and chorio-decidua, in particular at the cervical site. This gelatinous substance contained hyaluronan at a concentration of 3.0 microg/mg wtw. CONCLUSION: It is well established that prelabour fetal membranes are considerably stronger than postlabour fetal membranes. Two features may explain this; a weakening of the amnion combined with a separation of amnion and chorio-decidua. The biomechanical changes are consistent with the decrease in collagen and decorin, and the increase in hyaluronan and biglycan demonstrated in this study. The separation of the membranes is caused by the formation of a gelatinous substance, rich in hyaluronan. The results indicate that the biomechanical changes are not merely secondary to the stress of labour but that an active maturation process is involved.

Biglycan↗
PubMed1994

Current topic: the role of amniotic endothelin in human pregnancy.

Endothelin (ET), a potent vasoconstrictor peptide, was originally isolated from culture medium of porcine aortic endothelial cells. Subsequently, ET was also reported to be produced by non-vascular tissues, and to be involved in various biological phenomena in these tissues. Recently, a high concentration of ET-1-like immunoreactivity (ET-1-LI) was detected in human amniotic fluid. Amnion tissue also contained a large amount of ET-1-LI, and cultured amnion cells secreted large amounts of ET-1-LI. The major component of ET-1-LI in these samples was ET-1. Moreover, the expression of prepro-ET-1 mRNA was detected in both amnion tissue and cultured amnion cells, indicating that ET-1 in the amniotic fluid originated from amnion cells. In addition, specific receptors for ET were detected in myometrium, decidua vera, chorion laeve and placenta, by both ligand binding analysis and Northern blot analysis. These findings suggest that ET-1 secreted from amnion cells plays a physiological role in human pregnancy. In this paper, the regulation of ET-1 production and expression of ET-receptors in avascular human amnion tissue are reviewed. The possible importance of amniotic ET in human pregnancy is also discussed.

Amniotic Fluid↗
PubMed1990

Prenatal development of gonadotropin-releasing hormone receptors in the rat anterior pituitary.

The development of pituitary GnRH receptors was studied in the rat with in vitro and in vivo autoradiography. GnRH receptors were first seen in pituitary primordia of 13-day-old fetuses. The binding was specific and saturable and was abolished in the presence of 10 microM synthetic GnRH. To examine whether GnRH was available to the fetus, amnionic fluid was collected on days E 12-18. RIA analyses showed that GnRH levels in the amnionic fluid were low on days 12 and 13 (0-20 pM/ml) and rose to 225 pM/ml on day E 16 before they declined to 110 pM/ml on fetal day E 18. The highest levels of GnRH in the amnionic fluid on day E 16 coincided with the first appearance of immunoreactive LH cells, as determined by immunohistochemistry. Intravenous injection of 500 microliters amnionic fluid into pentobarbital-anesthetized adult rats caused a transient 40-60% increase in circulating serum LH in the recipient animal. To show that GnRH from the amnionic fluid has access to the developing pituitary, the 125I-labeled GnRH agonist Buserelin was injected into the amnionic fluid of 13-, 14-, and 15-day-old fetuses in the presence or absence of 10 microM unlabeled GnRH. Autoradiographic analysis of the fetal tissue indicated that the labeled GnRH agonist bound to specific receptors in the primordial pituitaries. The results suggest that the pituitary gonadotropes are differentiated before day E 13 because the expression of GnRH receptors is already an indication of cell determination. Since GnRH is present in the amnionic fluid in a biologically active form and can reach the fetal pituitary, it is concluded that GnRH may be an important factor determining the onset LH synthesis, but not the differentiation, of primordial pituitary cells.

Amniotic Fluid↗
PubMed2002

Apoptosis and related proteins during parturition in prostaglandin F receptor-deficient mice.

This study investigated whether apoptosis and related proteins are involved in parturition by comparative observation of FP-deficient mice without labor and wild type mice with vaginal delivery. We examined the expression of apoptosis, Fas, FasL, active caspase-3 and bcl-2 proteins in the amnion, placenta and decidua. DNA laddering in the amnion, placenta and decidua tissue did not significantly differ between FP-deficient and wild type mice on day 18 of pregnancy. Similar TUNEL staining results were found in all tissues of FP-deficient mice compared with those of wild type mice. A higher intensity of apoptotic cells was found in the decidua basalis. The index of TUNEL-positive cells were not significantly different in the amnion, placenta and decidua of FP-deficient mice compared with that of wild type mice on day 18 of pregnancy. Specific bands for Fas were clearly observed in the amnion, placenta and decidua tissue. FasL specific bands were observed in the placenta and decidua, but a few in amnion tissue. A great number of active caspase-3 specific bands were detected in decidua, while a few such bands were detected in the placenta and few bands in the amniotic tissue. Bands for bcl-2 were detected in the amnion, placenta and decidua tissue. The weakest band was in decidual tissue. Fas, FasL, active caspase-3, and bcl-2 specific bands did not show any significant differences between the two groups. These findings demonstrate that apoptosis, Fas, FasL, caspase-3, and Bcl-2 occur in mouse term placenta that is not involved in parturition.

Amnion↗
PubMedDate not supplied

Diffusion of dextrans and microspheres in the human amniotic basement membrane model.

The human amniotic basement membrane model was utilized to determine diffusion ratios of dextrans and beads according to size selectivity. Diffusion through both intact and denuded amnions was determined after 24 and 72 h. Four neutrally charged fluorescein isothiocyanate labeled dextrans, having molecular weights of 17,900, 42,000, 71,200 and 156,000, diffused through the amnion. The amnion functioned as a sieve in that the passage of dextrans was increasingly restricted as molecular weight increased. In contrast, uncharged latex microspheres (1.05 micron +/- 0.07 micron (SD] and fluorescent carboxylated microspheres (1.57 micron +/- 0.13 micron (SD] failed to pass through the amnion. Light and electron microscopy revealed no preformed channels through which the 1.05 micron microspheres could pass through the amnion. Statistical analysis of cross-sectional thickness of individual and similarly treated amnions (intact or denuded) showed a difference in thickness (P = 0.05).

Amnion↗
PubMed1993

Immunohistochemical localization of glucose transporters and insulin receptors in human fetal membranes at term.

The localization has been investigated of the isoforms GLUT1, GLUT3 and GLUT4 of glucose transporter proteins as well as of insulin receptors. Fetal membranes (n = 10) were examined by immunohistochemical methods at the light and electron microscopic levels using mono- and polyclonal antibodies. In all amnion epithelial cells, GLUT1 and GLUT3 antibodies were bound to the apical membrane. Very rarely the GLUT1 antibody also immunostained the basolateral membrane and reacted weakly with the endomembrane system and membranes of the lateral cell protrusions. Fibroblasts reacted with the antibodies against GLUT1, GLUT4 and insulin receptor, whereas they were labelled only in one case with GLUT3 antibody. Cytotrophoblast cells were only stained with antibodies against GLUT1 and GLUT3. Antibodies against GLUT4 only reacted with fibroblasts in the membranes. On amnion epithelial cells, weak immunoreactivity with insulin receptor antibodies was detected only at the electron microscopic level. The data indicate: (1) GLUT1 is located on all cells of the amnion, whereas GLUT3 is present in detectable amounts only on amnion epithelial cells and cytotrophoblast; (2) GLUT1 and GLUT3 on amnion epithelial cells are predominantly located on the apical surface; (3) GLUT4 and insulin receptors are not regularly expressed. We suggest that amnion epithelial cells cover their basal glucose requirements from the amniotic fluid and not from the maternal circulation.

Amnion↗
PubMed1989

Umbilical cord is the major source of prostaglandin E2 in the gestational sac during term labor.

The umbilical cord is an amniotic structure histologically resembling amnion lining the basal plate and reflected chorion. Prostaglandin E2 is secreted by amnion and is present in amniotic fluid. This study measured prostaglandin E2 production by amnion from all three locations to determine the relative contributions of prostaglandin E2 to amniotic fluid at term. Total surface areas and weights of umbilical cord, basal placental plate, and reflected chorionic amnion were measured in afterbirths from 20 normal patients delivered at term by elective repeat cesarean section before the onset of labor or vaginally after spontaneous onset of labor. Subsequently, 2 cm lengths of umbilical cord and 8 cm2 disks of basal placental plate and reflected chorionic amnion were incubated in perfusion chambers, and prostaglandin E2 production was measured by radioimmunoassay. Umbilical cord accounted for the least surface area (16% to 17%) but greatest tissue mass (75% to 76%). Both basal placental plate and reflected chorionic amnion increased prostaglandin E2 production 2.3-fold and 4.1-fold, respectively, after labor versus before labor, whereas umbilical cord prostaglandin E2 output was unchanged. However, umbilical cord accounted for 66% and 44% of the total prostaglandin E2 output before labor (697 +/- 169 ng/hr) versus (1201 +/- 380 ng/hr) after labor. Thus, of the three amniotic locations, umbilical cord represents the principal site of prostaglandin E2 production within the gestational sac.

Amnion↗
PubMed1990

The role of fetal membranes in regulating production, transport, and metabolism of prostaglandin E2 during labor.

The level of prostaglandin E2, a potent stimulator of myometrial contractions, increases in amniotic fluid during labor. We tested whether amniotic prostaglandin E2 crosses the reflected fetal membranes to the myometrium during labor. Disks of amnion, chorion-decidua, and amnion chorion-decidua from 10 term pregnancies before labor and nine vaginal deliveries after normal spontaneous labor were incubated in two-sided perfusion chambers in the absence or presence of 1 mumol/L indomethacin, or in the presence of a bolus of prostaglandin E2 and radiolabeled prostaglandin E2 added to the fetal chamber. Net amniotic prostaglandin E2 increased significantly (p = 0.003) after labor. However, there was no significant difference in prostaglandin E2 concentrations on the maternal side of amnion chorion-decidua between labor conditions. High-pressure liquid chromatography identified the transport of intact tritiated prostaglandin E2 across amnion but not across chorion-decidua or amnion chorion-decidua in which prostaglandin E2 was completely metabolized. Because of 15-hydroxyprostaglandin dehydrogenase activity in chorion, we suggest that amniotic prostaglandin E2 transport to the myometrium is impaired. Thus the increase in prostaglandin E2 production that occurs in amnion tissue after labor commences does not result in an increase in prostaglandin E2 concentration on the myometrial side of the fetal membranes in the in vitro preparation.

Adult↗
PubMed1984

Estrogen and progesterone receptor and hormone levels in human myometrium and placenta in term pregnancy.

Estradiol and progesterone receptors in the myometrium, decidua, placenta, chorion, and amnion of eight women who underwent elective cesarean section at term were determined by means of an exchange assay. The hormone levels in the peripheral plasma and cytosol of these tissues were measured by radioimmunoassays. Maternal plasma and the placenta had high concentrations of estradiol and progesterone, with the placenta having 12 times more progesterone than in maternal plasma but only half the concentrations of estradiol in maternal plasma. The decidua and placenta had detectable levels of cytosol and nuclear estradiol receptors, but the myometrium had no measurable cytosol estradiol receptors, whereas the chorion and amnion had neither cytosol nor nuclear estradiol receptors. However, the chorion and amnion had significantly higher concentrations of estradiol in the cytosol than those in the decidua and myometrium. Only the decidua and myometrium had cytosol and nuclear progesterone receptors, but the placenta, amnion, and chorion had neither cytosol nor nuclear progesterone receptors. In contrast, progesterone hormone levels were significantly higher in the placenta, amnion, and chorion than in the decidua and myometrium. The findings indicate that, in the term pregnant uterus, (1) the placenta, amnion, and chorion are rich in progesterone, estradiol, and nuclear estradiol receptors but have no progesterone receptors, (2) the decidua and myometrium have nuclear estradiol and progesterone receptors, and (3) the myometrium has a higher progesterone/estradiol ratio than that of the peripheral plasma, thus suggesting a highly progesterone-dominated uterus.

Amnion↗
PubMedDate not supplied

Developmental expression of vascular endothelial growth factor and its receptors in ovine placenta and fetal membranes.

OBJECTIVES: Vascularity of the surface of the placenta in humans and of the placenta and fetal membranes in several species including sheep is an important determinant of intramembranous absorption of amniotic fluid. Our previous studies have shown that the total blood vessel surface area in ovine amnion and chorion increases with advancing gestation. Vascular endothelial growth factor (VEGF) is a potent angiogenic and permeability factor and is found to be expressed in the ovine placenta and fetal membranes. To investigate the role of VEGF in maintaining the absorptive function of the intramembranous microvessels, the present study was undertaken to determine the gestational change in gene expression of VEGF and its receptors, kinase insert domain-containing receptor (KDR) and fms-like tyrosine kinase (Flt-1), in ovine placenta, chorion, and amnion. METHODS: Total RNA was extracted from placental cotyledon, chorion, and amnion of ovine fetuses at 60-140 days of gestation. The relative abundance of VEGF, KDR, and Flt-1 mRNA was determined by Northern blot analysis, and VEGF molecular forms expressed were identified by reverse transcriptase polymerase chain reaction. The gestational changes in mRNA levels of VEGF and its receptors were analyzed by regression analysis. RESULTS: In ovine placenta, chorion, and amnion, VEGF mRNA levels increased significantly from 60 to 140 days. The major VEGF molecular form expressed in these tissues was VEGF164, whereas VEGF120, VEGF144, and VEGF188 were present at lower levels. In the placenta, KDR was the primary VEGF receptor expressed, although Flt-1 was also detected at very low levels. In the amnion and chorion, KDR was the only receptor expressed. A gestational-dependent change in VEGF receptor expression was not observed in the placenta and membranes. CONCLUSIONS: The increase in VEGF gene expression with advancing gestation in the amnion and chorion where KDR is expressed suggests that VEGF and its receptor are important determinants of vascularity and permeability, and thus exchange capacity, of the intramembranous pathway.

Amnion↗
PubMed1961

A comparative study of the growth, nutrition, and metabolism of the primary and the transformed human cells in vitro.

The growth characteristics, nutritional requirements, and metabolic activities of primary explants (chiefly amnion) and transformed human cells (derived from amnion, conjunctiva, and cervical cancer) were compared. Major differences observed are as follows: 1. Five types of human tissue when freshly explanted in vitro were found to have varying but limited growth potential in contrast to the autonomous and apparently unlimited growth of the transformed cell. 2. A requirement for an external source of inositol or glutamine could not be demonstrated for the slowly multiplying primary amnions in contrast to the rapidly growing transformed cells which degenerated in the absence of either metabolite. 3. Unlike the transformed cells, the primary amnion cells fixed an insignificant amount of CO(2); when infected by the Coxsackie virus, however, a requirement for CO(2) in the formation of more virus became readily demonstrable. 4. The biggest difference in the incorporation of various C(14) substrates occurred in the nucleic acid fractions. 5. The difference in the incorporation of glycine into the protein fraction was very small. 6. The utilization of glucose, ribose, and xylose as well as the oxidation of lactic acid into CO(2) by the primary amnion cells occurred at a rate much lower than that of the transformed cells. 7. The cytotoxic dose of 5-fluorodeoxyuridine for the primary amnion was at least 100 times that of the transformed cells although growth inhibition was readily demonstrated at 0.1 microg. per ml. for both types of cells. Based on these observations, several hypotheses are put forth for consideration.

Amnion↗
PubMed1998

Gene expression of plasminogen activation cascade components in human term gestational tissues with labour onset.

The plasminogen activation cascade is thought to play a critical role in labour-associated remodelling events, such as fetal membrane rupture and placental separation. The aim of this study was to quantify, by Northern analysis, the gene expression of urokinase plasminogen activator (UPA), urokinase receptor (UPAR) and plasminogen activator inhibitor type-2 (PAI-2) in human gestational tissues. Amnion, choriodecidua and placenta were collected from women before, during and after spontaneous-onset labour at term. The expression of UPAR mRNA was significantly (P < 0.05) increased in amnion tissue during and after labour and delivery, compared with the before-labour group. In contrast, UPAR gene expression in choriodecidua and placenta was not significantly altered in association with labour onset. PAI-2 mRNA expression was also significantly (P < 0.05) increased in amnion after labour. No statistically significant differences were observed in choriodecidua or placenta PAI-2 mRNA with labour onset. Neither was any significant effect of labour status on UPA mRNA identified in any of the tissues examined. This study is the first to describe a significant increase in UPAR and PAI-2 gene expression in human amnion tissue with labour. These data are consistent with the hypothesis that, during labour, up-regulation of UPAR expression in amnion serves to localize active UPA at the cell surface, thereby increasing proteolytic activity in fetal membranes. Increased PAI-2 in amnion after labour may provide a regulatory 'switch' to cease further proteolysis in this tissue type. In conclusion, the data obtained support the proposal that the plasminogen activation cascade contributes to the rupture of fetal membranes during active labour.

Amnion↗
PubMed2003

Apoptosis induced by influenza virus-hemagglutinin stimulation may be related to fluctuation of cellular oxidative condition.

Primary cultivated amnion epitherial cells prepared from amnion tissue of human fetal membrane (Amnion-cells) were stimulated with influenza virus-hemagglutinin (IV-HA), fractionated from a commercialized IV-HA vaccine by DEAE Sephacel column chromatography. From 72-96 h after stimulation, chromosomal DNA fragmentation and the appearance of in situ TUNEL stained-positive cells were revealed. Amnion-cell DNA fragmentation was inhibited in the presence of glycophorin A or C purified from the human erythrocyte membrane fraction, but not inhibited with free N-acetyl-neuraminic acid. RT-PCR and Western blotting analysis showed that anti-oxidative enzymes were altered with incubation period, accompanied by the expression of the cellular oxidative stress-related caspase cascade. Pre-stimulation of Amnion-cells with hemin, a heme oxygenase-1 inducer, significantly attenuated IV-HA induced DNA fragmentation. It is concluded that IV-HA induces apoptosis in Amnion-cells, and that this apoptotic induction may be facilitated by certain sialoglycoproteins on the cell surface, and is related to changes in the intracellular redox condition.

Amnion↗