PSIII-9 Supplementation of beta-carotene or fatty acids alters production of prostaglandins in bovine endometrial epithelial cells

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 288-289
Author(s):  
Allison R Harman ◽  
Rebecca Swanson
Keyword(s):  
Epithelial Cells ◽  
Antioxidant Properties ◽  
Beta Carotene ◽  
Endometrial Cells ◽  
Prostaglandin Production ◽  
Uterine Environment ◽  
Free Media ◽  
Endometrial Epithelial Cells

Abstract Differential prostaglandin secretion from the bovine endometrium can be used as a marker for an embryotropic or embryotoxic uterine environment. Beta-carotene has antioxidant properties and is the precursor for retinol, which has been shown to improve early embryonic development in vivo and in vitro. Furthermore, dietary fatty acid supplementation, specifically eicosapentaenoic acid (EPA) and decosahexaenoic acid (DHA) has been shown to alter prostaglandin production. The objective of this study was to determine prostaglandin production of endometrial cells following treatment with beta-carotene, EPA, or DHA. Bovine endometrial epithelial cells were treated for 24 hours with serum-free media supplemented with either 10 µM beta-carotene, 10 µM EPA, 10 µM DHA or ethanol (>1% volume/volume) vehicle control. After treatment, concentrations of PGE2 and PGF2a were analyzed in media via commercially available ELISA kits. Concentrations and ratios of prostaglandins were analyzed via ANOVA using the mixed procedure in SAS version 9.4. Beta-carotene treatment decreased PGE2 (P < 0.0001) and PGF2a (P = 0.0003) concentrations in media compared to controls. However, the ratio of PGE2:PGF2a was not different (P = 0.1203) between beta-carotene and controls. DHA treatment decreased PGE2 (P < 0.0001) concentrations in media but did not alter (P = 0.1079) PGF2a concentrations in media compared to controls. The ratio of PGE2:PGF2a was not different (P = 0.6343) between DHA and controls. EPA treatment did not alter (P = 0.1503) PGE2 concentrations in media compared to controls. Conversely, PGF2a concentrations were decreased (P = 0.0088) in media treated with EPA compared to controls. Therefore, the ratio of PGE2:PGF2a was increased (P = 0.0116) between EPA versus controls. These studies demonstrate that in vitro supplementation of EPA may alter the endometrial synthesis of prostaglandins to be more embryotropic. Therefore, EPA may be therapeutic for in vivo trials to influence the early uterine environment.

Increased mRNA expression of selected pro-inflammatory factors in inflamed bovine endometrium in vivo as well as in endometrial epithelial cells exposed to Bacillus pumilus in vitro

2016 ◽  
Vol 28 (7) ◽  
pp. 982 ◽  
Author(s):  
Martina A. Gärtner ◽  
Sarah Peter ◽  
Markus Jung ◽  
Marc Drillich ◽  
Ralf Einspanier ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mrna Expression ◽  
Bacillus Pumilus ◽  
Cultured Cells ◽  
Inflammatory Factors ◽  
Endometrial Cells ◽  
Subclinical Endometritis ◽  
Endometrial Epithelial Cells

Endometrial epithelium plays a crucial role in the first immune response to invading bacteria by producing cytokines and chemokines. The aim of this study was to investigate the first inflammatory response of the endometrium in vivo and in vitro. Gene expression of several pro-inflammatory factors and Toll-like receptors (TLR2, -4, -6) was determined in endometrial cytobrush samples obtained from healthy cows and cows with clinical or subclinical endometritis. Endometrial epithelial cells were co-cultured with an isolated autochthonous uterine bacterial strain Bacillus pumilus. Total RNA was extracted from in vivo and in vitro samples and subjected to real-time reverse transcription polymerase chain reaction. CXC ligands (CXCL) 1/2 and CXC chemokine receptor (CXCR) 2 mRNA expression was higher in cows with subclinical endometritis and CXCL3 mRNA expression was higher in cows with clinical endometritis compared with healthy cows. B. pumilus induced cell death of epithelial cells within 24 h of co-culturing. The presence of B. pumilus resulted in significantly higher mRNA expression of interleukin 1α (IL1A), IL6, IL8, CXCL1–3 and prostaglandin–endoperoxide synthase 2 in co-cultured cells compared with untreated controls. The maximum increase was mainly detected after 2 h. These results support the hypothesis that bacterial infection of endometrial cells might induce prompt synthesis of pro-inflammatory cytokines resulting in a local inflammatory reaction.

Treatment of Intrauterine Adhesions with Human Amnion Mesenchymal Stromal Cells (hAMSCs) on Promoting Endometrial Regeneration and Repair Via Notch Signaling Pathway Regulation

2020 ◽  
Author(s):  
Jie Yu ◽  
Wenwen Zhang ◽  
Jiayue Huang ◽  
Yating Gou ◽  
Congcong Sun ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Notch Signaling ◽  
Signaling Pathway ◽  
Notch Signaling Pathway ◽  
Intrauterine Adhesions ◽  
Epithelial Markers ◽  
Qrt Pcr ◽  
Endometrial Epithelial Cells

Abstract Background: Human amniotic mesenchymal stem cells(hAMSCs) can repair and improve the damaged endometrium which its aplastic disorder is the main reason for intrauterine adhesions(IUAs).Methods: We conducted in vivo and in vitro experiments. In vivo experiments: 45 female Sprague-Dawley(SD) rats were involved and randomized equally into Sham group, IUA group, Estradiol(E2) group, hAMSCs group, and E2 + hAMSCs group. The effect of hAMSCs and E2 only or combined was evaluated by Hematoxylin-eosin(HE) and Masson staining. The expression of epithelial markers and key proteins of Notch signaling pathway by Immunohistochemistry. In vitro experiments: Firstly, the hAMSCs cells were taken and divided into control group and induced group in which hAMSCs were differentiated into endometrial epithelial cells in induced microenvironment, and extracted their RNA respectively. The expression of epithelial markers and Notch1 messenger RNA (mRNA) was detected by Real-time quantitative polymerase chain reaction(qRT-PCR). and the changes in expression position of Notch intracellular domain(NICD) and expression amount of target gene, hairy enhancer of split 1(Hes1) were detected by Immunofluorescence. Then, Activated and inhibited the Notch signaling pathway while induction, and detected mRNA expression of hAMSCs epithelial markers by quantitative real-time polymerase chainreaction (qRT-PCR) respectively and detected hAMSCs cell cycle by flow cytometric. Results:This study showed that hAMSCs alone or combined with E2 could promote endometrial repair, and Notch signaling pathway a great role in it. And otherwise, the activation or habitation of Notch signaling pathway determines whether hAMSCs could differentiate into endometrial epithelial cells or not.Conclusion: we concluded that activate the Notch signaling pathway promote the differentiation of hAMSCs into endometrial epithelial cells, and further treat IUAs.

scholarly journals Mitochondrial dysfunction induced by high estradiol concentrations in endometrial epithelial cells

2019 ◽  
Author(s):  
Chia-Hung Chou ◽  
Shee-Uan Chen ◽  
Chin-Der Chen ◽  
Chia-Tung Shun ◽  
Wen-Fen Wen ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mitochondrial Dysfunction ◽  
Embryo Implantation ◽  
Ros Production ◽  
Vitro Fertilization ◽  
Equine Chorionic Gonadotropin ◽  
In Vivo Effects ◽  
Endometrial Epithelial Cells

Abstract Context A supraphysiological estradiol (E2) concentration after ovarian stimulation is known to result in lower embryo implantation rates in in vitro fertilization (IVF). Endometrial epithelial cells (EECs) apoptosis occurs after the stimulation with high E2 concentrations, and mitochondria play important roles in cell apoptosis. Objective To investigate the mitochondrial function in EECs after the stimulation with high E2 concentrations. Materials and Methods Human EECs were purified and cultured with different E2 concentrations (10-10, 10-9, 10-8, 10-7 M) in vitro, in which 10-7 M is supraphysiologically high. Eight-week-old female mouse endometrium was obtained 5.5 days after the injection of 1.25 IU or 20 IU equine chorionic gonadotropin (eCG), roughly during the embryo implantation window, to examine the in vivo effects of high E2 concentrations on mouse EECs. Results In vivo and in vitro experiments demonstrated decreased mitochondrial DNA contents and ATP formation after EECs were stimulated with supraphysiologically high E2 concentrations than those stimulated with a physiologic E2 concentration. Less prominent immunofluorescence mitochondrial staining, fewer mitochondria number under electron microscopy, lower JC-1 aggregate/monomer ratio, and greater reactive oxygen species (ROS) production were found after EECs were stimulated with supraphysiologically high E2 concentrations. The high E2-induced ROS production was reduced when EECs were pretreated with N-acetyl-cysteine (NAC) in vitro, but remained unchanged after the pretreatment with coenzyme Q10. Conclusion High E2 concentrations increase extra-mitochondrial ROS production in EECs and subsequently result in mitochondrial dysfunction.

Cell Growth Factor and Estrogen Inducing Menstrual Blood-Derived Stem Cells (MenSC) Differentiate into Endometrial Epithelial Cells

2022 ◽  
Vol 12 (3) ◽  
pp. 659-664
Author(s):  
Wei Li ◽  
Tieying Shan ◽  
Jianping Shi ◽  
Zexian Fu ◽  
Shujing Qi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Epithelial Cells ◽  
Menstrual Blood ◽  
Control Group ◽  
Cell Staining ◽  
Artificial Induction ◽  
Uterine Environment ◽  
Group Control Group ◽  
Endometrial Epithelial Cells

Extracted MenSC (Menstrual blood-derived stem cells) from female menstrual blood. Added various exogenous factors in-vitro and simulated the female uterine environment to observe how to make MenSC differentiation into Endometrial epithelial cells by artificial induction. MenSCs were divided into 4 groups: 2.5×10−5 mol/L E group, 1.613 nmol/L EGF group, 2.5×10−5 mol/L E+1.613 nmol/L EGF group, control Group (only MenSCs); the relevant indicators of the experiment includes cell staining and Western Blot to detect CK and VIM protein content; RT-PCR to detect CK-19 mRNA and VIM mRNA. The cell staining results showed that E+EGF group had significant differentiation in 7 days and 14 days. CK-19mRNA of E+EGF group was significantly higher than other groups, and the EGF group expression was obviously higher than that of E group, and VIMmRNA expression is opposite to that. The protein expression had the similar performance. MenSC can differentiate into endometrial epithelial cells after induced by E and EFG; and the co-culture of E and EFG can achieve better differentiation, which proves their work together in MenSC differentiate towards endometrial epithelial cells.

Ultrastructure of human blastocyst-endometrial interactions in vitro

Reproduction ◽  
2000 ◽  
pp. 337-350 ◽  
Author(s):  
U Bentin-Ley ◽  
T Horn ◽  
A Sjogren ◽  
S Sorensen ◽  
J Falck Larsen ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Plasma Membranes ◽  
Lateral Displacement ◽  
Syncytium Formation ◽  
Surface Epithelium ◽  
Stromal Compartment ◽  
Endometrial Cells ◽  
Trophoblastic Cells ◽  
Endometrial Epithelial Cells

The interactions of seven human blastocysts with cultured endometrial cells were investigated by light microscopy and transmission electron microscopy. Trophoblastic-endometrial contact was observed at the lateral border of endometrial epithelial cells where trophoblast and endometrial epithelial cells shared apical junctional complexes and desmosomes. The first sign of penetration was invasion of a trophoblastic cytoplasmic protrusion between endometrial epithelial cells. In broad contact areas, lateral displacement of endometrial epithelial cells and formation of a peripheral pseudostratified epithelium were observed. When trophoblastic cells were interposed fully among endometrial epithelial cells, they formed a penetration cone and appeared to dislodge endometrial epithelial cells from the stromal compartment. A single penetration cone only was found in each specimen. Endometrial or trophoblastic degeneration was not observed. Formation of multinucleate (>/= three nuclei per cell) trophoblast cells was not observed, but many cells displayed areas with abrupt disappearance of well-defined plasma membranes, which is indicative of syncytium formation. In this study, adhesion and penetration occurred at the same time. The human blastocysts penetrated the endometrial surface epithelium by intrusive penetration. Epithelial penetration was achieved primarily by cellular syncytiotrophoblast-like cells and the first indications of syncytium formation were observed simultaneously with penetration of the epithelium.

scholarly journals P-244. Expression of oestrogen receptors in endometrial epithelial cells in vivo and in vitro

1999 ◽  
Vol 14 (Suppl_3) ◽  
pp. 263-263
Author(s):  
E.M. Tuckerman ◽  
M.E. Page ◽  
T.C. Li ◽  
S.M. Laird
Keyword(s):  
Epithelial Cells ◽  
Oestrogen Receptors ◽  
Endometrial Epithelial Cells

scholarly journals Minireview: Putting Physiology Back into Estrogens' Mechanism of Action

Endocrinology ◽  
2011 ◽  
Vol 152 (12) ◽  
pp. 4481-4488 ◽  
Author(s):  
Robert D. Koos
Keyword(s):  
Epithelial Cells ◽  
Cultured Cells ◽  
Hypoxia Inducible Factor ◽  
Estrogen Receptor Α ◽  
Proliferative Effect ◽  
High Oxygen Level ◽  
Endothelial Growth Factor ◽  
Endometrial Epithelial Cells

After decades of research, the mechanism by which estrogens stimulate the proliferation of epithelial cells in the endometrium and mammary gland, and in the carcinomas that arise in those tissues, is still not understood. Cells do not proliferate in response to 17β-estradiol (E2) alone, and although it is widely recognized that growth factors play a role in E2's proliferative effect, exactly how they are involved is unclear. It has long been known that the proliferation of endometrial epithelial cells is preceded by dramatic increases in blood flow and microvascular permeability, filling the subepithelial stroma with plasma and the proteins it contains, such as IGF-I, which is known to synergize with E2 in the induction of cell proliferation. The hyperpermeability is caused by vascular endothelial growth factor (VEGF), which is rapidly induced by E2, via the transcription factors hypoxia-inducible factor 1 and estrogen receptor α, in luminal epithelial cells in vivo. As we recently showed, VEGF is also strongly induced in endometrial cancer cells in vitro when excessive degradation of hypoxia-inducible factor 1α, caused by the abnormally high oxygen level to which cultured cells are exposed, is prevented. Putting these facts together, we now propose a new model of E2-induced proliferation in which VEGF-induced vascular hyperpermeability plays an essential role. E2 first induces the expression by endometrial epithelial cells of VEGF, which then acts in a paracrine manner to induce interendothelial cell gaps in subepithelial blood vessels, through which plasma and the proteins therein enter the adjacent stroma. Plasma carries even more E2, which circulates bound to proteins, and IGF-l, which together drive epithelial cells completely through the cell cycle.

scholarly journals FBXO18/Circrna211/Mir-431/CSF1 Axis Regulates the Establishment of Endometrial Receptivity Though Activating MAPK and CSF1R/PI3K/AKT/Mtor Pathways in Dairy Goats

2021 ◽  
Author(s):  
Lichun Yang ◽  
Xiaorui Liu ◽  
Lei Zhang ◽  
Danni Li ◽  
Guili Li ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Regulatory Network ◽  
Target Genes ◽  
Endometrial Receptivity ◽  
Physiological Status ◽  
Dairy Goats ◽  
In Vivo Models ◽  
Endometrial Epithelial Cells

Abstract Background: Endometrial epithelial cells proliferation and secretion of various cytokines have a strong impact on the formation of receptive endometrium, which is known as a physiological status that allows an activated embryo to attach to the endometrium for a limited time. Circular RNAs and miRNAs can be involved in the dynamic physiological changes of endometrium by regulating relevant functional target genes in the uterus. Our work presented here with the ultimate purpose of revealing the latent molecular mechanism of FBXO18/circRNA211/miR-431/CSF1 axis in the establishment of endometrial receptivity of dairy goats.Results: In vitro, we found a regulatory network of FBXO18/circRNA211/miR-431/CSF1 in goat endometrial epithelial cells that circRNA211 severed as a sponge for miR-431, resulting in weakening the inhibition of miR-431 on target genes CSF1 and FBXO18. FBXO18/circRNA211/miR-431/CSF1 axis promoted the proliferation through regulating the key proteins of Ras, Raf, MEK, ERK in MAPK pathway via CCK-8, EdU, flow cytometry and Western blot assays. Furthermore, FBXO18/circRNA211/miR-431/CSF1 axis activated the phosphorylation of key proteins PI3K, AKT and mTOR in PI3K-mTOR pathway by CSF1R, thereby promoting the establishment of endometrial receptivity. In vivo models, mice injected with miR-431 agomir showed that the endometrial thickness and the number of pinopodes were significantly decreased by HE staining and scanning electron microscope. Immunohistochemistry results showed that VEGF and OPN proteins were down-regulated and MUC1 protein was up-regulated under the treatment of miR-431 agomir. Further study demonstrated that miR-431 inhibited embryo implantation by impeding the establishment of endometrial receptivity.Conclusion: Ultimately, our study revealed a regulatory mechanism of FBXO18/circRNA211/miR-431/CSF1 axis in goat endometrial epithelial cells. This circRNA/miRNA/mRNA regulatory network presented here in vitro and in vivo models may provide a novel insight into the potentially regulating endometrium biological functions and promoting the formation of endometrium receptivity.

103 ESHERICHIA COLI LIPOPOLYSACCHARIDE STIMULATES PROLIFERATION OF BOVINE UTERINE EPITHELIAL CELLS

2014 ◽  
Vol 26 (1) ◽  
pp. 165 ◽  
Author(s):  
Y. Guo ◽  
M. Chanrot ◽  
P. Reinaud ◽  
G. Charpigny ◽  
O. Sandra ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Untreated Control ◽  
Initial Number ◽  
Endometrial Cells ◽  
Cell Numbers ◽  
Esherichia Coli ◽  
Passage Number ◽  
Number Of Cells ◽  
Endometrial Epithelial Cells

Lipopolysaccharide (LPS) is a component of the outer membrane of gram-negative bacteria involved in the pathogenic processes leading to postpartum metritis and endometritis in dairy cows. It also causes inflammation of the endometrium and implantation failures in many animal species. An increase in cell proliferation by LPS has been reported in human epithelial and immune cells (Martin et al. 2000 J. Immunol. 165, 139–147) but not in endometrial cells. The aim of this study was to characterise the proliferative response of bovine endometrial cells following exposure to Escherichia coli LPS. In vitro cultures of bovine endometrial epithelial cells (EEC) and fibroblasts were performed following collection of bovine endometrium at the slaughterhouse. Endometrial epithelial cells and fibroblasts were separated before primary culture (Charpigny et al. 1999 J. Reprod. Fertil. 117, 315–324). On passages 4 to 6, EEC issued from 5 cows (total 25 replicates) were challenged with 2, 4, 8, 12, 16, or 24 μg mL–1 of LPS. At the time of challenge and 72 h later, the numbers of attached cells were counted for control and LPS-treated groups. The variation in cell numbers after the challenge was calculated as the ratio of the number of LPS-treated cells minus the number of untreated control cells to the number of untreated control cells. The variation in cell numbers over time was analysed by ANOVA (SAS 9.1, PROC GLM; SAS Institute Inc., Cary, NC, USA) following arcsin transformation of percentages. The effect of passage number, initial number of cells at the time of challenge, treatment group, and corresponding interactions were included in the model. A significant increase in cell numbers was observed for cells treated with 2, 4, 8, and 12 μg mL–1 of LPS (+21 ± 5%, +30 ± 5%, +41 ± 4%, and +14 ± 5% over the control, respectively; P ≤ 0.001), whereas a nonsignificant effect was observed for 16 and 24 μg mL–1 of LPS (+4 ± 5%, and –5 ± 14%, respectively). Effects of passage number and initial number of cells at the time of challenge were nonsignificant and no interactions of those factors with treatment effects were observed. For fibroblasts, preliminary results (2 replicates from 2 cows) suggest that the response to LPS is much less important than for EEC (+3.6 and +1.3% for 2 μg mL–1 of LPS; +5.8 and +4% for 8 μg mL–1 of LPS). These findings indicate that E. coli LPS stimulates proliferation of bovine endometrial epithelial cells, and that the effect of LPS is dose dependent but not linear under the range of concentrations tested.

scholarly journals Estrogen degrades Scribble in endometrial epithelial cells through E3 ubiquitin ligase HECW1 in the development of diffuse adenomyosis

2019 ◽  
Author(s):  
Zhixing Jin ◽  
Haiou Liu ◽  
Congjian Xu
Keyword(s):  
Epithelial Cells ◽  
3D Culture ◽  
Expression Level ◽  
Eutopic Endometrium ◽  
Estrogen Production ◽  
Tissue Specimens ◽  
Endometrial Epithelial Cells

Abstract Despite its prevalence and the severity of symptoms, little is known about the pathogenesis and etiology of adenomyosis. In previous studies, the protein expression level of the polarity protein Scribble in the eutopic endometrium of patients with adenomyosis was found to be significantly decreased; however, little is known about its regulatory mechanism. In consideration of the important role of supraphysiologic estrogen production in the endometrium in the development of adenomyosis, we analyzed the effect and mechanism of estrogen on the expression of Scribble in vivo and in vitro. Firstly, we found Scribble was down-regulated in eutopic endometrium and negatively related with aromatase P450 in Tamoxifen-induced adenomyosis. Then, we established a 3D culture of primary endometrial epithelial cells and used found that estrogen could disrupt apical-basal polarity of endometrial glandular epithelial cells. Based on the following experiments and GEO datasets screening, we found estrogen regulates the expression level of Scribble by HECW1 through ubiquitination of Scribble protein. At last, we verified the expression of Scribble, HECW1 and aromatase P450 in eutopic endometrium of human and mouse specimens and found the expression of HECW1 and aromatase P450 was significantly increased, while the expression of Scribble was significantly downregulated. Furthermore, a positive correlation was found between HECW1 and aromatase P450, while a negative correlation was found between HECW1 and Scribble in human clinical tissue specimens. Therefore, our research may provide a new understanding of the pathogenesis of adenomyosis.

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