Altered release of endothelin‐1,2 and thromboxane B<sub>2</sub> from trophoblastic cells in pre‐eclampsia

Červar, M.; Kainer, Franz; Jones, Carolyn J.P.; Desoyé, Gernot

doi:10.1046/j.1365-2362.1996.87238.x

Cited by 22 publications

(12 citation statements)

References 35 publications

(56 reference statements)

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“…Also, these concentrations encompass the physiological range described during normal pregnancy, preeclampsia, and within placental tissues. 3,4,36 Immunocytochemistry Cells were cultured 4 hours, 16 hours, or 72 hours as above on Lab-Tek Chamber Slide System (Nunc, Naperville, IL) using 4-well Permanox slides, and cells were fixed for 20 minutes in absolute methanol, rinsed with 3 changes of cold phosphate-buffered saline (PBS), and immunostained as described below.The purity of the cultures, after cell attachment for 4 hours and rinsing as above, was analyzed using the trophoblast-specific monoclonal mouse antihuman cytokeratin 7 antibody (Biomeda Corporation, Foster City, CA; 1:50) and detected using a secondary donkey antimouse IgG (H+L; 2 mg/mL; Cytotrophoblasts and syncytiotrophoblasts were distinguished by co-localization of surface membranes by antidesmosomal staining and nuclei by ToPro (Sigma) staining. We detected a cytoplasmic epithelial marker of apoptosis, the cleavage product of cytokeratin 18 intermediate filaments (cyt 18), using the M30 CytoDeath Antibody (Roche Applied Science, Indianapolis, IN; 1:100) imaged by an Alexa Fluor 555-tagged mouse IgG2b labeling kit (Molecular Probes).…”

Section: Trophoblast Isolation and Culturementioning

confidence: 99%

Endothelin-1 Attenuates Apoptosis in Cultured Trophoblasts From Term Human Placentas

Cervar-Zivkovic

Barton

et al. 2007

Reprod. Sci.

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The authors test the hypothesis that endothelin-1 (ET-1) modulates apoptosis in human term trophoblasts. Primary cultures of cytotrophoblasts from term human placentas (n = 5) were cultured for 16 hours total or 24 hours prior to harvest at 72 hours in atmospheres of <1%, 8%, and 20% oxygen, in the presence of 10% serum, ET-1 (1-100 pmol/mL), both, or neither. The apoptotic cleavage products of poly-ADP-ribose polymerase and cytokeratin 18 filaments were quantified by Western analysis and immunocytochemistry. The expression of BAD, pBAD-serine 112, p53, and 2 isoforms of MDM2 were quantified by immunoblotting, and endothelin A and B receptors were analyzed by immunocytochemistry. Compared to vehicle control, increasing concentrations of ET-1 reduce by 3- to 6-fold the level of apoptosis in cytotrophoblasts exposed to serum-free conditions at 20% oxygen. Similarly, syncytiotrophoblast cultures grown for 24 hours without serum in 100 pmol/mL ET-1 show a 3-fold lower level of apoptosis compared with vehicle control. ET-1 significantly reduces apoptosis in cultures exposed to 20% oxygen but not in cultures exposed to 8% or 1% oxygen. The effect of ET-1 on apoptosis in 20% oxygen is accompanied by reduced p53 expression and is correlated with enhanced expression of endothelin B receptor, compared to cultures in 8% or 1% oxygen. ET-1 reduces apoptosis in cultured human trophoblasts, and this finding suggests a role for ET-1 in protecting trophoblasts against injury.

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Section: Trophoblast Isolation and Culturementioning

confidence: 99%

Endothelin-1 Attenuates Apoptosis in Cultured Trophoblasts From Term Human Placentas

Cervar-Zivkovic

Barton

et al. 2007

Reprod. Sci.

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“…Villi from women with these conditions typically exhibit diminished villus formation, prominent cytotrophoblasts, enhanced syncytial knot formation, and apoptosis (6,7). An increase in TX synthesis has been demonstrated in villi (8,9) and trophoblasts (10,11) from preeclamptic women compared with healthy control subjects. Importantly, FGR (12) and preeclampsia (13) are associated with elevated TXA 2 levels in the circulation of pregnant women.…”

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confidence: 99%

Thromboxane A2 Limits Differentiation and Enhances Apoptosis of Cultured Human Trophoblasts

et al. 2001

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Prostanoids influence differentiation in diverse cell types. Altered expression of cyclooxygenase and prostaglandins has been implicated in the pathophysiology of placental dysfunction, which results in preeclampsia and fetal growth restriction. We hypothesized that prostanoids modulate differentiation and apoptosis in cultured human trophoblasts. Villous cytotrophoblasts were isolated from term human placentas and cultured in serumfree medium. The level of human chorionic gonadotropin was used as a marker of biochemical differentiation of primary trophoblasts, and syncytia formation was used as a marker of morphologic differentiation. Of the prostanoids tested, we found exposure to thromboxane A 2 hindered both biochemical and morphologic differentiation of cultured trophoblasts. As expected, human chorionic gonadotropin levels in the media were elevated in a concentration-dependent manner in the presence of the thromboxane synthase inhibitor, sodium furegrelate, or the thromboxane A 2 receptor blocker SQ 29,548. Furthermore, thromboxane A 2 enhanced trophoblast apoptosis, determined using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining, cell morphology, and a concentrationdependent increase in p53 expression. We conclude that thromboxane A 2 hinders differentiation and enhances apoptosis in cultured trophoblasts from term human placenta. We speculate that thromboxane may contribute to placental dysfunction by restricting differentiation and enhancing apoptosis in human trophoblasts. The function of the human placenta depends on multinucleated, terminally differentiated syncytiotrophoblasts. This epithelium is in direct contact with maternal blood and regulates maternal-fetal exchange of micronutrients and gases. The syncytiotrophoblast arises from fusion of relatively undifferentiated, mitotically active cytotrophoblasts. This process involves morphologic and biochemical differentiation. Morphologic differentiation is defined by fusion of mononucleated cytotrophoblasts with adjacent syncytium (1). Biochemical differentiation is characterized by production of hormones such as hCG and human placental lactogen (2-4). Preeclampsia and FGR are associated with trophoblast hypoxia and placental dysfunction (5). Villi from women with these conditions typically exhibit diminished villus formation, prominent cytotrophoblasts, enhanced syncytial knot formation, and apoptosis (6, 7). An increase in TX synthesis has been demonstrated in villi (8, 9) and trophoblasts (10, 11) from preeclamptic women compared with healthy control subjects. Importantly, FGR (12) and preeclampsia (13) are associated with elevated TXA 2 levels in the circulation of pregnant women. Although the effect of prostanoids on vascular reactivity is well known (14), recent studies indicate that prostanoids also regulate proliferation, differentiation, and apoptosis in many cell types (15-18). We tested the hypothesis that prostanoids influence the differentiation of cultured trophoblasts from term human placenta. We found ...

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“…HLA-class-I-expressing cells were stained using W6/32.HL (mouse IgG; 1:20; Sera-Lab), directed against the monomorphic moiety of β 2 m-associated HLA class I molecules (HLA-A, -B, -C, -G), which are not expressed in all villous syncytio-and cytotrophoblast cells, but appear strongly in all other cellular components of placental tissue including extravillous cytotrophoblast, due to their expression of HLA-G (Cervar et al 1996). However, extravillous cytotrophoblast cells are rarely present in term placentas and, therefore, in the following HLA-class-I-expressing cells are referred to as "non-trophoblast" cells.…”

Section: Methodsmentioning

confidence: 99%

“…Tumor necrosis factor-alpha (TNF-α) strongly inhibits trophoblast cell motility and adhesion in vitro (Todt et al 1996). In some diseases, like preeclampsia, unknown factors induce the reduced trophoblast attachment and syncytialization (Pijnenborg et al 1996) without affecting the hCG-β release in vitro (Cervar et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Paracrine regulation of distinct trophoblast functions in vitro by placental macrophages

Červar

Blaschitz

Dohr

et al. 1999

Cell and Tissue Research

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In view of the accumulating evidence for paracrine mechanisms regulating trophoblast function, we tested the hypothesis that placental macrophages affect trophoblast activity in a paracrine fashion. Trophoblast was isolated from 17 term placentas (-IP). One aliquot of cells was further immunopurified (+IP) using an HLA class I antibody. This increased the proportion of trophoblast (+IP >97%; -IP approximately 70%) as identified by rigorous immunocytochemistry. Most (approximately 70%) non-trophoblast cells in -IP were macrophages. The cells were cultured for 5 days with a daily medium change. In addition, +IP cells from seven placentas were cultured with lipopolysaccharide (LPS)-stimulated or -unstimulated macrophage-conditioned media. The concentrations of lactate, trophoblast-specific hormones, human chorionic gonadotropin-beta (hCG-beta) and human placental lactogen (hPL), of several prostanoids and of endothelin-1 and angiotensin II were determined in the culture media. The accumulated amounts of substances released into the culture media, corrected for the greater proportion of trophoblast in +IP cultures, were on average two- to threefold higher (hCG-beta: 18-fold) in +IP than in -IP, with the exception of endothelin-1,2 (no change), angiotensin II (-70%) and 6-keto-prostaglandin-F1alpha (-40%). [3H]leucine incorporation into the trichloroacetic acid (TCA)-precipitable pool measured on day 5 was twofold higher in +IP than in -IP. Addition of conditioned media reverted these changes. The data demonstrate that placental macrophages in culture affect trophoblast biosynthetic activity in a paracrine fashion. We conclude that macrophages are important regulators of trophoblast activity.

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Altered release of endothelin‐1,2 and thromboxane B₂ from trophoblastic cells in pre‐eclampsia

Cited by 22 publications

References 35 publications

Endothelin-1 Attenuates Apoptosis in Cultured Trophoblasts From Term Human Placentas

Endothelin-1 Attenuates Apoptosis in Cultured Trophoblasts From Term Human Placentas

Thromboxane A2 Limits Differentiation and Enhances Apoptosis of Cultured Human Trophoblasts

Paracrine regulation of distinct trophoblast functions in vitro by placental macrophages

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Altered release of endothelin‐1,2 and thromboxane B2 from trophoblastic cells in pre‐eclampsia

Cited by 22 publications

References 35 publications

Endothelin-1 Attenuates Apoptosis in Cultured Trophoblasts From Term Human Placentas

Endothelin-1 Attenuates Apoptosis in Cultured Trophoblasts From Term Human Placentas

Thromboxane A2 Limits Differentiation and Enhances Apoptosis of Cultured Human Trophoblasts

Paracrine regulation of distinct trophoblast functions in vitro by placental macrophages

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Altered release of endothelin‐1,2 and thromboxane B₂ from trophoblastic cells in pre‐eclampsia