Effect of glucose and oxygen deprivation on heme oxygenase expression in human chorionic villi explants and immortalized trophoblast cells

2003 ◽  
Vol 285 (6) ◽  
pp. R1453-R1460 ◽  
Author(s):  
S. D. Appleton ◽  
G. E. Lash ◽  
G. S. Marks ◽  
K. Nakatsu ◽  
J. F. Brien ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Protein Content ◽  
Heme Oxygenase ◽  
Glucose Concentration ◽  
Placental Tissue ◽  
Glucose Deprivation ◽  
First Trimester ◽  
Trophoblast Cells ◽  
Chorionic Villi ◽  
Low Glucose

Although hypoxia induces heme oxygenase (HO)-1 mRNA and protein expression in many cell types, recent studies in our laboratory using human placental tissue have shown that a preexposure to hypoxia does not affect subsequent HO enzymatic activity for optimized assay conditions (20% O2; 0.5 mM NADPH; 25 μM methemalbumin) or HO-1 protein content. One of the consequences of impaired blood flow is glucose deprivation, which has been shown to be an inducer of HO-1 expression in HepG2 hepatoma cells. The objective of the present study was to test the effects of a 24-h preexposure to glucose-deprived medium, in 0.5 or 20% O2, on HO protein content and enzymatic activity in isolated chorionic villi and immortalized HTR-8/SVneo first-trimester trophoblast cells. HO protein content was determined by Western blot analysis, and microsomal HO enzymatic activity was measured by assessment of the rate of CO formation. HO enzymatic activity was increased ( P < 0.05) in both placental models after 24-h preexposure to glucose-deficient medium in 0.5 or 20% O2. Preexposure (24 h) in a combination of low O2 and low glucose concentrations decreased the protein content of the HO-1 isoform by 59.6% ( P < 0.05), whereas preexposure (24 h) to low glucose concentration alone increased HO-2 content by 28.2% in chorionic villi explants ( P < 0.05). In this preparation, HO enzymatic activity correlated with HO-2 protein content ( r = 0.825). However, there was no correlation between HO-2 protein content and HO enzymatic activity in HTR-8/SVneo trophoblast cells preexposed to 0.5% O2 and low glucose concentration for 24 h. These findings indicate that the regulation of HO expression in the human placenta is a complex process that depends, at least in part, on local glucose and oxygen concentrations.

Effects of hypoxia on heme oxygenase expression in human chorionic villi explants and immortalized trophoblast cells

2003 ◽  
Vol 284 (3) ◽  
pp. H853-H858 ◽  
Author(s):  
S. D. Appleton ◽  
G. S. Marks ◽  
K. Nakatsu ◽  
J. F. Brien ◽  
G. N. Smith ◽  
...  
Keyword(s):  
Protein Expression ◽  
Protein Content ◽  
Heme Oxygenase ◽  
Protein Function ◽  
Cell Types ◽  
First Trimester ◽  
Trophoblast Cells ◽  
Human Trophoblast ◽  
Chorionic Villi ◽  
Enzymatic Function

Although hypoxia induces heme oxygenase (HO)-1 protein and mRNA expression in many cell types, hypoxia has also been shown to decrease HO-1 mRNA and protein expression. We tested the hypothesis that 24-h preexposure to hypoxia in human placental preparations suppresses HO protein expression and enzymatic function. Immortalized HTR-8/SVneo first-trimester trophoblast cells and explants of normal human chorionic villi (CV) from term placentas were cultured for 24 h in 1%, 5%, or 20% O2. HO protein levels were determined by Western blot analysis, and microsomal HO activity was measured. HO-2 protein content was decreased by 17% and 5% in human trophoblast cells after 24-h exposure to 1% and 5% O2, respectively, versus 20% O2. In contrast, HO-2 protein content in CV explants was unaffected by changes in oxygenation. HO-1 protein content, which was barely detectable in both biological systems, was not affected by changes in oxygenation. Similarly, HO enzymatic activity was unchanged in both preparations after 24-h exposure to 1%, 5%, or 20% O2. The above data do not support the hypothesis that hypoxia in the human placenta suppresses both HO protein content and HO protein function. The present observations reinforce the necessity to determine both HO protein expression and function.

Heme Oxygenase Expression in Selected Regions of Term Human Placenta

2003 ◽  
Vol 228 (5) ◽  
pp. 564-567 ◽  
Author(s):  
Brian E. McLaughlin ◽  
Gendie E. Lash ◽  
Graeme N. Smith ◽  
Gerald S. Marks ◽  
Kanji Nakatsu ◽  
...  
Keyword(s):  
Enzymatic Activity ◽  
Protein Content ◽  
Heme Oxygenase ◽  
Immunoblot Analysis ◽  
Chorionic Villi ◽  
Complementary Role ◽  
Optimized Conditions ◽  
Catalyzed Oxidation ◽  
Western Immunoblot Analysis ◽  
Normotensive Pregnancy

Carbon monoxide (CO), formed during heme oxygenase (HO)-catalyzed oxidation of heme, has been proposed to play a complementary role with nitric oxide in the regulation of placental hemodynamics. The objective of this study was to elucidate HO enzymatic activity and HO-1 (inducible) and HO-2 (constitutive) protein content in the microsomal subcellular fraction of homogenate of selected regions of placenta from normotensive and mild pre-eclamptic pregnancies. HO enzymatic activity was measured under optimized conditions by gas chromatography using CO formation as an index of activity, and HO-1 and HO-2 protein content were determined by Western immunoblot analysis. Microsomal HO activity in each of the four placental regions was not different between normotensive and mild pre-eclamptic pregnancies. Microsomal HO-2 protein content was not different between normotensive and mild pre-eclamptic pregnancies, whereas there was increased expression of microsomal HO-1 protein in chorionic villi and fetal membranes from pre-eclamptic pregnancy compared with normotensive pregnancy. Microsomal HO enzymatic activity correlated with HO-2, but not HO-1, protein content.

Use of endocervical trophoblasts in noninvasive prenatal testing for common aneuploidies

2020 ◽  
Vol 19 (4) ◽  
pp. 75-80
Author(s):  
E.V. Musatova ◽  
◽  
M.V. Kapustina ◽  
M.E. Minzhenkova ◽  
Zh.G. Markova ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Prenatal Testing ◽  
First Trimester ◽  
Dna Probes ◽  
Fish Analysis ◽  
Trophoblast Cells ◽  
Noninvasive Prenatal Testing ◽  
Chorionic Villi ◽  
Trimester Screening ◽  
Reliable Methods

The article presents an experience of isolation of trophoblast cells from cervical samples of pregnant women and demonstrates a possibility of determining aneuploid cells in cervical samples using FISH analysis. Objective. To study methods of effective isolation and reliable detection of trophoblasts for genetic pathology analysis. Patients and methods. The participants of the study were three pregnant women, who according to the findings of the combined first trimester screening test were referred to the group with a high risk for fetal aneuploidy. The terms of gestation varied from 12 to 14 wks. Immunocytochemical detection of trophoblast cells was performed with FITC-labelled HLA-G antibodies. FISH was performed with the use of locus-specific DNA probes on chromosomes 13/21 and a DNA probe on chromosome 18 centromere. The chromosome set of a fetus was verified by the results of standard cytogenetic analysis of semi-direct chromosome preparations from cytotrophoblast cells obtained by chorionic villi biopsy. Results. HLA-G-positive cells were found in all examined cytological specimens. FISH analysis with the use of DNA probes on chronomomes, whose trisomies are compatible with live birth, detected aneuploid cells in all three cases. Conclusion. The use of cervical trophoblasts for noninvasive collection of information about the genetic status of the developing fetus at present needs further study of effective and reliable methods of their isolation, detection and analysis. Key words: aneuploidy, throphoblast cells, NIPT, prenatal diagnosis, HLA-G

scholarly journals Downregulation of A1 and A2B adenosine receptors in human trisomy 21 mesenchymal cells from first-trimester chorionic villi

2012 ◽  
Vol 1822 (11) ◽  
pp. 1660-1670 ◽  
Author(s):  
Stefania Gessi ◽  
Stefania Merighi ◽  
Angela Stefanelli ◽  
Prisco Mirandola ◽  
Alessandra Bonfatti ◽  
...  
Keyword(s):  
Adenosine Receptors ◽  
Trisomy 21 ◽  
First Trimester ◽  
Mesenchymal Cells ◽  
Chorionic Villi

scholarly journals Intraperitoneal Microdialysis as a Monitoring Method in the Intensive Care Unit

2014 ◽  
Vol 99 (6) ◽  
pp. 729-733 ◽  
Author(s):  
Tasiopoulos Konstantinos ◽  
Komnos Apostolos ◽  
Paraforos Georgios ◽  
Tepetes Konstantinos
Keyword(s):  
Intensive Care Unit ◽  
Intensive Care ◽  
Glucose Concentration ◽  
Repeated Measures ◽  
Statistical Significance ◽  
Monitoring Method ◽  
P Values ◽  
Repeated Measures Analysis ◽  
Low Glucose ◽  
Surgical Condition

Abstract Studies on surgical patients provide some evidence of prompt detection of enteric ischemia with microdialysis. The purpose of the study was to measure intraperitoneal microdialysis values (glucose, glycerol, pyruvate, and lactate) in patients hospitalized in an intensive care unit (ICU) with an underlying abdominal surgical condition and to correlate these values with patients' outcomes. Twenty-one patients, 10 female, were enrolled in the study. The intraperitoneal metabolite values were measured for 3 consecutive days, starting from the first day of ICU hospitalization. Descriptive and inferential statistics were performed. The t-test, repeated measures analysis, Holm's test, and a logistic regression model were applied. Level of statistical significance was set at P = 0.05. Mean age of participants was 68.10 ± 8.02 years old. Survivors exhibited statistically significantly higher glucose values on day 3 (6.61 ± 2.01 against 3.67 ± 1.62; P = 0.002). Mean lactate/ pyruvate (L/P) values were above 20 (35.35 ± 27.11). All non-survivors had a mean three day L/P values greater than 25.94. Low L/P values were related to increased survival possibilities. High microdialysis glucose concentration, high L/P ratio and low glucose concentration were the major findings during the first three ICU hospitalization days in non-survivors. Intraperitoneal microdialysis may serve as a useful tool in understanding enteric ischemia pathophysiology.

TSH/miR‐17‐5p/ZNF367 axis is related to spontaneous abortion in patients with TSH above 2.5 mIU/L

2021 ◽  
Author(s):  
Yang Yang ◽  
Jiashu Li ◽  
Yingying Zhou ◽  
Wen Dai ◽  
Weiping Teng ◽  
...  
Keyword(s):  
Pregnant Women ◽  
Spontaneous Abortion ◽  
Biological Function ◽  
Zinc Finger Protein ◽  
First Trimester ◽  
Cell Counting ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
Chorionic Villi ◽  
Increased Risk

Elevated thyroid stimulating hormone (TSH) is associated with an increased risk of spontaneous abortion (SA); however, the associated mechanism remains unclear. This study aimed to investigate the expression of microRNAs (miRNAs) and pathogenesis in the chorionic villi of TSH > 2.5 mIU/L-related SA patients. The chorionic villi were collected from pregnant women in the first trimester with TSH > 2.5 mIU/L with or without SA, as well as TSH < 2.5 mIU/L with or without SA to determine the level of miRNA expression. Differentially expressed miRNAs were confirmed by qRT-PCR in a total of 92 subjects. Cell counting kit-8 (CCK8), wound healing, transwell assays, and Western blotting were used to measure cellular biological functions and related protein in HTR-8/SVneo cells. The potential mechanisms were determined using a Luciferase reporter assay and rescue experiment. Compared with normal pregnant women, miR-17-5p was decreased and zinc finger protein 367 (ZNF367) was upregulated in the chorionic villi of TSH > 2.5 mIU/L-related SA patients. Using HTR-8/SVneo cells, we demonstrated that elevated TSH inhibited miR-17-5p expression, as well as trophoblast migration and invasion. The overexpression of miR-17-5p targeted and inhibited ZNF367 expression promoting the biological function of trophoblasts. Further studies confirmed that ZNF367 interference partially reversed the biological function of the miR-17-5p inhibitor on HTR-8/SVneo cells. Taken together, our results showed that miR-17-5p promoted biological function of trophoblasts by suppressing ZNF367.

Effect of the oxygen tension on the expression and function of Galβ1-3GalNAc disaccharide in the first trimester trophoblast cells

Placenta ◽  
2011 ◽  
Vol 32 ◽  
pp. S333-S334
Author(s):  
L. Ermini ◽  
A. Spagnoletti ◽  
N. Bechi ◽  
S. Aldi ◽  
J. Bhattacharjee ◽  
...  
Keyword(s):  
Oxygen Tension ◽  
First Trimester ◽  
Trophoblast Cells ◽  
And Function

Relationship between proliferation and glucose metabolism in primary cultures of rabbit proximal tubules

1990 ◽  
Vol 259 (3) ◽  
pp. C455-C461 ◽  
Author(s):  
M. J. Tang ◽  
R. L. Tannen
Keyword(s):  
Protein Content ◽  
Cellular Proliferation ◽  
Primary Cultures ◽  
Glucose Consumption ◽  
Cell Number ◽  
Glycolytic Enzyme ◽  
Proximal Tubules ◽  
Low Glucose ◽  
Lactate Formation ◽  
Glucose Restriction

Primary cultures of rabbit proximal tubules, which revert to a glycolytic profile as reflected by increased activity of pyruvate kinase (PK) paralleled by increased glucose consumption and lactate formation, were utilized to explore the relationship between glycolytic metabolism and proliferation. Tubules placed in serum-free, hormonally defined Dulbecco's modified Eagle's medium with 5 mM glucose exhibited logarithmic growth beginning on day 3 in culture. The increase in PK activity lagged approximately 1 day behind, suggesting that the reversion to glycolysis is a consequence of rather than a prerequisite for cellular proliferation. Tubules cultured in 0.5 mM as contrasted with 25 mM glucose exhibited heightened proliferation reflected by an increase in protein content and cell number on day 5 in culture. The heightened proliferation was accompanied by increased PK activity. On day 9, after confluency had been achieved, no differences in protein content or PK activity were detected between tubules cultured in different glucose concentrations. These findings indicate that a low glucose concentration is mitogenic for renal proximal tubules and that the proliferative process in some fashion up-regulates the activity of the glycolytic enzyme PK. Furthermore, because accelerated growth proceeds in the presence of glucose restriction, the energy from glycolysis is not required for the proliferative process.

scholarly journals Blocking Epidermal Growth Factor Receptor Signaling in HTR-8/SVneo First Trimester Trophoblast Cells Results in Dephosphorylation of PKBα/AKT and Induces Apoptosis

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
J. Bolnick ◽  
L. Albitar ◽  
L. L. Laidler ◽  
R. Abdullah ◽  
K. K. Leslie
Keyword(s):  
Protein Phosphorylation ◽  
Global Analysis ◽  
Growth Factor Receptor ◽  
First Trimester ◽  
Extravillous Trophoblast ◽  
Signaling Proteins ◽  
Trophoblast Cells ◽  
Novel Technology ◽  
Epidermal Growth ◽  
Growth Factor Receptor Signaling

We identified a major peptide signaling target of EGF/EGFR pathway and explored the consequences of blocking or activating this pathway in the first trimester extravillous trophoblast cells, HTR-8/SVneo. A global analysis of protein phosphorylation was undertaken using novel technology (Kinexus Kinetworks) that utilizes SDS-polyacrylamide minigel electrophoresis and multi-lane immunoblotting to permit specific and semiquantitative detection of multiple phosphoproteins. Forty-seven protein phosphorylation sites were queried, and the results reported based on relative phosphorylation at each site. EGF- and Iressa-(gefitinib, ZD1839, an inhibitor of EGFR) treated HTR-8/SVneo cells were subjected to immunoblotting and flow cytometry to confirm the phosphoprotein screen and to assess the effects of EGF versus Iressa on cell cycle and apoptosis. EGFR mediates the phosphorylation of important signaling proteins, including PKBα/AKT. This pathway is likely to be central to EGFR-mediated trophoblast survival. Furthermore, EGF treatment induces proliferation and inhibits apoptosis, while Iressa induces apoptosis.

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