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.

Mitochondrial Dysfunction Induced by High Estradiol Concentrations in Endometrial Epithelial Cells

2019 ◽  
Vol 105 (1) ◽  
pp. 126-135 ◽  
Author(s):  
Chia-Hung Chou ◽  
Shee-Uan Chen ◽  
Chin-Der Chen ◽  
Chia-Tung Shun ◽  
Wen-Fen Wen ◽  
...  
Keyword(s):  
Mitochondrial Dysfunction ◽  
Embryo Implantation ◽  
Ros Production ◽  
Vitro Fertilization ◽  
Endometrial Epithelial Cell ◽  
Dna Contents ◽  
Equine Chorionic Gonadotropin ◽  
In Vivo Effects

Abstract Context A supraphysiological estradiol (E2) concentration after ovarian stimulation is known to result in lower embryo implantation rates in in vitro fertilization. Endometrial epithelial cell (EEC) 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, 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 numbers under electron microscopy, lower 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide 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 in vitro, but remained unchanged after the pretreatment with coenzyme Q10. Conclusion High E2 concentrations increase extramitochondrial ROS production in EECs and subsequently result in mitochondrial dysfunction.

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 Identification of a Novel Role for Endothelins within the Oviduct

Endocrinology ◽  
2010 ◽  
Vol 151 (6) ◽  
pp. 2858-2867 ◽  
Author(s):  
Myoungkun Jeoung ◽  
Sungeun Lee ◽  
Hee-kyung Hawng ◽  
Yong-Pil Cheon ◽  
Youn Kyung Jeong ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Embryo Development ◽  
Early Embryo ◽  
Receptor Subtypes ◽  
Early Embryo Development ◽  
Vitro Fertilization ◽  
Protein Superfamilies ◽  
Endothelin 3

Endothelins were first identified as potent vasoactive peptides; however, diversity in the biological function of these hormones is now evident. We have identified a novel role for endothelins: a requirement for these peptides within the oviduct during fertilization and/or early embryo development. In vivo, treatment after ovulation with a dual endothelin receptor antagonist (tezosentan) decreased the number of two-cell embryos that could be collected from within the oviducts. In vitro fertilization experiments showed that gamete viability and their ability to fertilize were not affected by treatment with this antagonist, suggesting that the effect observed in vivo was mediated by the oviduct itself. Expression of mRNA for all three isoforms of the endothelins and both receptor subtypes was detectable within the oviduct. Expression of mRNA for endothelin-3 was regulated by gonadotropins in epithelial cells of the oviduct and increased specifically within the isthmus of this structure. Immunostaining revealed localization of both endothelin receptors A and B to the columnar epithelial cells within the oviduct, suggestive of a local role for endothelins in the regulation of epithelial function and ultimately oviductal secretions. A microarray analysis revealed three likely endothelin-regulated protein networks for future analysis: the TGFβ, IL-10, and CCAAT/enhancer-binding protein superfamilies. Overall, these results suggest a novel and requisite role for endothelins within the oviduct during fertilization and/or early embryo development.

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.

scholarly journals Versican V1 in human endometrial epithelial cells promotes BeWo spheroid adhesion in vitro

Reproduction ◽  
2019 ◽  
pp. 53-64 ◽  
Author(s):  
Yumiko Miyazaki ◽  
Akihito Horie ◽  
Hirohiko Tani ◽  
Masashi Ueda ◽  
Asuka Okunomiya ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Chondroitin Sulfate ◽  
Cell Line ◽  
Human Embryo ◽  
Embryo Implantation ◽  
Cancer Cell Line ◽  
Promoting Effect ◽  
Ishikawa Cells ◽  
Endometrial Epithelial Cells

The endometrium extracellular matrix (ECM) is essential for embryo implantation. Versican, a large chondroitin sulfate proteoglycan that binds hyaluronan and forms large ECM aggregates, can influence fundamental physiological phenomena, such as cell proliferation, adhesion and migration. The present study investigated the possible role of versican in human embryo implantation. Versican V1 expression and secretion in human endometrial epithelial cells (EECs) was most prominent in the mid-secretory phase. Versican expression in EECs significantly increased after treatment with estrogen and progesterone, but not by estrogen alone. We also established versican V1-overexpressing Ishikawa (endometrial cancer cell line) cells (ISKW-V1), versican V3-overexpressing (ISKW-V3) and control GFP-overexpressing (ISKW-GFP) Ishikawa cells. By the in vitro implantation model, the attachment ratio of BeWo (choriocarcinoma cell line) spheroids to the monolayer of ISKW-V1, but not of ISKW-V3, was found significantly enhanced compared with attachment to the ISKW-GFP monolayer. The conditioned medium derived from ISKW-V1 (V1-CM) also promoted the attachment of BeWo spheroids to the ISKW monolayer. However, this attachment-promoting effect was abolished when V1-CM was pretreated with chondroitinase ABC, which degrades chondroitin sulfate. Therefore, out of the ECM components, versican V1 may facilitate human embryo implantation.

In vitro Stimulation of Granulosa Cells by a Combination of Different Active Ingredients of Unkei-to

2004 ◽  
Vol 32 (04) ◽  
pp. 569-578 ◽  
Author(s):  
Wen-Shu Sun ◽  
Atsushi Imai ◽  
Keiko Tagami ◽  
Michiyo Sugiyama ◽  
Tatsuro Furui ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Response Curves ◽  
Vitro Fertilization ◽  
Human Granulosa Cells ◽  
Estradiol Secretion ◽  
Paeoniae Radix ◽  
In Vivo Effects ◽  
Stimulation Of

Unkei-to is widely used in traditional Japanese herbal medicine for its ovulation-inducing effect. In the present study, we investigated the in vivo effects of Unkei-to and its compounds on the steroidogenesis and cytokine secretion in human granulosa cells. Unkei-to stimulated the secretions of 17β-estradiol and progesterone from highly luteinized granulosa cells obtained from in vitro fertilization patients; the stimulated effect on estradiol secretion occurred with 0.3 μg/ml, while a significant effect on progesterone secretion was obtained at 10 μg/ml. The Unkei-to stimulation of estradiol secretion could be accounted for by the effects of its ingredients, Shakuyaku (paeoniae radix, Paeonia lactiflora Pallas) and Keihi (cinnamomi cortex, Cinnamomum cassia Blume); while dose response curves for Unkei-to and Keihi to induce progesterone production were superimposable. Exposure of the cells to Unkei-to caused dose-dependent increases in the concentrations of interleukin (IL)-1β, IL-6 and IL-8 in the culture medium. Similar results were obtained when cells were incubated with the ingredient Ninjin (ginseng radix, Panax ginseng C.A. Meyer), but not Shakuyaku and Keihi. These results indicate that Unkei-to has direct stimulatory effects on human granulosa cells to stimulate the steroidogenesis and secretion of cytokines (IL-1β, IL-6 and IL-8). The various beneficial actions of Unkei-to on the ovary may result from a combination of different ingredient herbs with different stimulatory effects on both steroidogenesis and the ovulatory process within the ovary, as well as stimulatory effect on the hypothalamus-pituitary axis.

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