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1.
Int J Mol Sci ; 22(16)2021 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-34445100

RESUMO

Endometriosis is a common gynecological disorder characterized by ectopic growth of endometrium outside the uterus and is associated with chronic pain and infertility. We investigated the role of the long intergenic noncoding RNA 01133 (LINC01133) in endometriosis, an lncRNA that has been implicated in several types of cancer. We found that LINC01133 is upregulated in ectopic endometriotic lesions. As expression appeared higher in the epithelial endometrial layer, we performed a siRNA knockdown of LINC01133 in an endometriosis epithelial cell line. Phenotypic assays indicated that LINC01133 may promote proliferation and suppress cellular migration, and affect the cytoskeleton and morphology of the cells. Gene ontology analysis of differentially expressed genes indicated that cell proliferation and migration pathways were affected in line with the observed phenotype. We validated upregulation of p21 and downregulation of Cyclin A at the protein level, which together with the quantification of the DNA content using fluorescence-activated cell sorting (FACS) analysis indicated that the observed effects on cellular proliferation may be due to changes in cell cycle. Further, we found testis-specific protein kinase 1 (TESK1) kinase upregulation corresponding with phosphorylation and inactivation of actin severing protein Cofilin, which could explain changes in the cytoskeleton and cellular migration. These results indicate that endometriosis is associated with LINC01133 upregulation, which may affect pathogenesis via the cellular proliferation and migration pathways.


Assuntos
Endometriose/genética , Endométrio/patologia , Células Epiteliais/patologia , RNA Longo não Codificante/genética , Adulto , Linhagem Celular , Proliferação de Células , Endometriose/patologia , Endométrio/citologia , Endométrio/metabolismo , Células Epiteliais/citologia , Células Epiteliais/metabolismo , Feminino , Humanos , Pessoa de Meia-Idade , Regulação para Cima , Adulto Jovem
2.
Theriogenology ; 173: 221-229, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34399386

RESUMO

Glucocorticoids (GCs) are known to play an important role in maintaining basal and stress-related homeostasis by interacting with endocrine mediators and prostaglandins (PGs). Although a growing body of evidence shows that GCs exert their regulatory action at a multitude of sites in the reproductive axis through corticosteroid receptors, little is known about the direct role of cortisol, an active form of GCs, in the equine endometrium. Thus, the study aimed to determine the effect of cortisol on PGF2α synthesis in the endometrial tissue and cells in vitro. In Exp.1, the immunolocalization and the expression of the glucocorticoid receptor (GCR) in the endometrium throughout the estrous cycle were established. In Exp. 2 and 3, the effects of cortisol on PGF2α secretion and transcripts associated with the arachidonic acid (AA) cascade in endometrial tissues, and cells were defined. Endometrial tissues obtained from the early, mid, and late luteal phases and the follicular phase of the estrous cycle were exposed to cortisol (100, 200, and 400 nM) for 24 h. Endometrial epithelial and stromal cells (early phase of estrous cycle) were exposed to cortisol (100 nM) for 24 h. Then, PGF2α secretion and transcripts associated with the AA cascade (PLA2G2A, PLA2G4A, PTGS2, and PGFS) were assessed. GCR was expressed in the cytoplasm and the nucleus in the luminal and glandular epithelium as well as in the stroma. Endometrial GCR protein abundance was up-regulated at the late luteal phase compared to the mid-luteal phase of the estrous cycle. Cortisol dose-dependently decreased PGF2α secretion, PLA2G2A and PLA2G4A transcripts in endometrial tissues. Additionally, cortisol treatment decreased PGF2α secretion from endometrial epithelial and stromal cells. Moreover, it affected PLA2G2A, PLA2G4A, and PTGS2 transcripts in endometrial stromal cells. These findings suggest that cortisol suppresses the synthesis of PGF2α by affecting the AA cascade in the equine endometrium during the estrous cycle.


Assuntos
Dinoprosta , Hidrocortisona , Animais , Ácido Araquidônico/metabolismo , Ácido Araquidônico/farmacologia , Dinoprosta/metabolismo , Dinoprosta/farmacologia , Dinoprostona/metabolismo , Endométrio/metabolismo , Feminino , Cavalos , Hidrocortisona/metabolismo , Redes e Vias Metabólicas
3.
Life Sci ; 283: 119854, 2021 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-34332980

RESUMO

AIMS: Cell adhesion molecule 1 (CADM1) mediates interepithelial adhesion and is upregulated in crowded epithelial monolayers. This study aimed to examine CADM1 expression in the human endometrium of proliferative and secretory phases, and its transcriptional regulation in terms of estrogen stimuli and higher cellularity. MAIN METHODS: CADM1 immunohistochemistry was conducted on endometrial tissues from women in their 40s and adult mice subcutaneously injected with estradiol following ovariectomy. Dual-luciferase reporter assays were conducted using human endometrial HEC-50B and HEC-1B cells and reporter plasmids harboring the human CADM1 3.4-kb promoter and its deleted and mutated forms. Cells were transfected with estrogen receptor α cDNA and reporter plasmids, and treated with estradiol before luciferase activity measurement. KEY FINDINGS: Immunohistochemistry revealed that CADM1 was clearly expressed on the lateral membranes of the simple columnar glandular cells in the proliferative phase, but not in the secretory phase, from both women and the mouse model. The glandular cell density increased two-fold in the proliferative phase. Reporter assays identified three Sp1-binding sites as estradiol-responsive elements in the proximal region (from -223 to -84) of the transcription start site (+1) in HEC-50B cells. When the cell culture was started at eight-fold higher cell density, the CADM1 3.4-kb promoter was transactivated at a two-fold higher level in HEC-50B cells. This cell density effect was not detected for the CADM1 2.3-kb or 1.6-kb promoter. SIGNIFICANCE: Two (proximal and distal) promoter regions are suggested to function additively to transactivate CADM1 in endometrial glandular cells that crowd in the proliferative phase.


Assuntos
Molécula 1 de Adesão Celular/biossíntese , Proliferação de Células , Endométrio/metabolismo , Células Epiteliais/metabolismo , Regulação da Expressão Gênica , Adulto , Animais , Molécula 1 de Adesão Celular/genética , Linhagem Celular Tumoral , Estrogênios/farmacologia , Feminino , Humanos , Camundongos
4.
Mol Med Rep ; 23(5)2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34227673

RESUMO

The estrogen 17ß­estradiol has been proven to serve an indispensable role in the occurrence and development of adenomyosis (ADS). The let­7a/Lin28B axis can control cell proliferation by acting as a tumor­inhibiting axis in numerous types of cancer. However, its role in ADS remains unknown. The present study aimed i) to elucidate the role of let­7a in regulating the proliferation of human uterine junctional zone (JZ) smooth muscle cells (SMCs) in ADS, ii) to evaluate whether 17ß­estradiol modifies the expression levels of let­7a and Lin28B in JZ SMCs in ADS, and iii) to establish how 17ß­estradiol affects the function of the let­7a/Lin28B axis in the proliferation of JZ SMCs in ADS. A total of 36 premenopausal women with ADS were enrolled as the experimental group and 34 women without ADS were recruited as the control group. Reverse transcription­quantitative PCR was used to evaluate the expression level of let­7a, and western blotting was performed to determine the Lin28B expression levels. Lentiviral null vector, let­7a overexpression lentiviral vector GV280 and let­7a inhibition lentiviral vector GV369 were used to infect cells to alter the expression of let­7a for further functional experiments. 17ß­estradiol and Cell Counting Kit­8 assays were conducted to determine how 17ß­estradiol affects the function of the let­7a/Lin28B axis in the proliferation of JZ SMCs in ADS. The results demonstrated that let­7a was downregulated and Lin28B was upregulated in the JZ SMCs of ADS compared with the control cells (P<0.0001). Moreover, a lower expression of let­7a led to faster proliferation of JZ SMCs (P<0.05), and 17ß­estradiol affected the let­7a/Lin28B axis to accelerate the proliferation of JZ SMCs in ADS (P<0.05). These data suggested that 17ß­estradiol collaborates with the let­7a/Lin28B axis to affect the development of ADS.


Assuntos
Adenomiose/tratamento farmacológico , Proliferação de Células/efeitos dos fármacos , Estradiol/farmacologia , MicroRNAs/metabolismo , Miócitos de Músculo Liso/efeitos dos fármacos , Proteínas de Ligação a RNA/metabolismo , Útero/efeitos dos fármacos , Adenomiose/genética , Adenomiose/metabolismo , Adulto , Proliferação de Células/genética , Endométrio/metabolismo , Feminino , Humanos , MicroRNAs/genética , Pessoa de Meia-Idade , Miócitos de Músculo Liso/metabolismo , Miométrio/metabolismo , Cultura Primária de Células , Proteínas de Ligação a RNA/genética , Transdução de Sinais/efeitos dos fármacos , Útero/metabolismo
5.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201586

RESUMO

The molecular mechanism underlying embryonic implantation is vital to understand the correct communications between endometrium and developing conceptus during early stages of pregnancy. This study's objective was to determine molecular changes in the uterine endometrial proteome during the preimplantation and peri-implantation between 9 days (9D), 12 days (12D), and 16 days (16D) of pregnant Polish Large White (PLW) gilts. 2DE-MALDI-TOF/TOF and ClueGOTM approaches were employed to analyse the biological networks and molecular changes in porcine endometrial proteome during maternal recognition of pregnancy. A total of sixteen differentially expressed proteins (DEPs) were identified using 2-DE gels and MALDI-TOF/TOF mass spectrometry. Comparison between 9D and 12D of pregnancy identified APOA1, CAPZB, LDHB, CCT5, ANXA4, CFB, TTR upregulated DEPs, and ANXA5, SMS downregulated DEPs. Comparison between 9D and 16D of pregnancy identified HP, APOA1, ACTB, CCT5, ANXA4, CFB upregulated DEPs and ANXA5, SMS, LDHB, ACTR3, HP, ENO3, OAT downregulated DEPs. However, a comparison between 12D and 16D of pregnancy identified HP, ACTB upregulated DEPs, and CRYM, ANXA4, ANXA5, CAPZB, LDHB, ACTR3, CCT5, ENO3, OAT, TTR down-regulated DEPs. Outcomes of this study revealed key proteins and their interactions with metabolic pathways involved in the recognition and establishment of early pregnancy in PLW gilts.


Assuntos
Implantação do Embrião/fisiologia , Endométrio/metabolismo , Prenhez/metabolismo , Proteínas/metabolismo , Animais , Feminino , Gravidez , Proteínas/análise , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Suínos
6.
FASEB J ; 35(8): e21784, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34252231

RESUMO

The human endometrium undergoes cycle-dependent changes and is only receptive to an implanting blastocyst within a narrow window of 2-4 days in the mid-secretory phase. Such functional changes require delicate interplay between a diversity of factors including cytokines and signaling pathways. The Notch signaling pathway members are expressed in human endometrium. We have previously demonstrated that Notch ligand Jagged1 (JAG1) localizes in the endometrial luminal epithelium (LE) and is abnormally reduced in infertile women during receptivity. However, the functional consequences of reduced JAG1 production on endometrial receptivity to implantation of the blastocyst are unknown. This study aimed to determine the role of JAG1 in regulating endometrial receptivity in humans and mice. Knockdown of JAG1 in both primary human endometrial epithelial cells and Ishikawa cells significantly reduced their adhesive capacity to HTR8/SVneo (trophoblast cell line) spheroids. We confirmed that in human endometrial epithelial cells, JAG1 interacted with Notch Receptor 3 (NOTCH3) and knockdown of JAG1 significantly reduced the expression of Notch signaling downstream target HEY1 and classical receptivity markers. Knockdown of Jag1 in mouse LE significantly impaired blastocyst implantation. We identified ten genes (related to tight junction, infertility, and cell adhesion) that were differentially expressed by Jag1 knockdown in LE in mice. Further analysis of the tight junction family members in both species revealed that JAG1 altered the expression of tight junction components only in mice. Together, our data demonstrated that JAG1 altered endometrial epithelial cell adhesive capacity and regulated endometrial receptivity in both humans and mice likely via different mechanisms.


Assuntos
Implantação do Embrião , Endométrio/metabolismo , Proteína Jagged-1/metabolismo , Transdução de Sinais , Adulto , Animais , Linhagem Celular , Feminino , Humanos , Proteína Jagged-1/genética , Camundongos
7.
DNA Cell Biol ; 40(7): 998-1008, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34115954

RESUMO

Transcripts of uncertain coding potential (TUCP) are part of long noncoding RNAs, which include short open reading frames and could be translated into small peptides. In recent years, a growing number of TUCPs has been implicated in multiple biological activities, such as embryogenesis and transcriptional regulation. However, the abundance of TUCPs and their roles in goat endometrium during pregnancy recognition (day 16) remain undocumented. In this study, bioinformatics analyses were conducted to identify the differentially expressed (DE) TUCPs between pregnant animals and corresponding nonpregnant controls. A total of 5551 TUCPs were identified; 114 TUCPs were DE in goat endometrium, of which 74 TUCPs were upregulated in pregnant endometrium, whereas 40 TUCPs were downregulated. The related genes of TUCP were predicted by using coexpression and colocalization methods. In summary, 419 genes were predicted by colocalization, and 9464 genes were predicted by coexpression. The kyoto encyclopedia of genes and genomes (KEGG) and gene ontology (GO) analysis showed that TUCPs, which are highly expressed in pregnant endometrium, were mainly associated with endometrial remodeling, nutrient synthesis, and transportation. However, TUCPs that were lowly expressed in pregnant endometrium were mainly associated with immune tolerance, which is necessary for the protection and development of the embryo in the uterus. These findings may be used for the comparative analysis of TUCP transcripts in endometrium and assist in the selection of applicable candidate genes associated with embryo implantation for further functional analyses.


Assuntos
Implantação do Embrião/genética , Endométrio/metabolismo , RNA Longo não Codificante/genética , Animais , Sequência de Bases/genética , Embrião de Mamíferos , Endométrio/fisiologia , Feminino , Expressão Gênica/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica/genética , Cabras/genética , Gravidez/genética , RNA Longo não Codificante/metabolismo , RNA Mensageiro/genética , Análise de Sequência de RNA/métodos , Transcriptoma/genética , Sequenciamento Completo do Exoma/métodos
8.
Commun Biol ; 4(1): 651, 2021 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-34140633

RESUMO

Assessment of the endometrium often necessitates a biopsy, which currently involves an invasive, transcervical procedure. Here, we present an alternative technique based on deriving organoids from menstrual flow. We demonstrate that organoids can be derived from gland fragments recovered from menstrual flow. To confirm they faithfully reflect the in vivo state we compared organoids derived from paired scratch biopsies and ensuing menstrual flow from patients undergoing in vitro fertilisation (IVF). We demonstrate that the two sets of organoids share the same transcriptome signature, derivation efficiency and proliferation rate. Furthermore, they respond similarly to sex steroids and early-pregnancy hormones, with changes in morphology, receptor expression, and production of 'uterine milk' proteins that mimic those during the late-secretory phase and early pregnancy. This technique has wide-ranging impact for non-invasive investigation and personalised approaches to treatment of common gynaecological conditions, such as endometriosis, and reproductive disorders, including failed implantation after IVF and recurrent miscarriage.


Assuntos
Endométrio/citologia , Menstruação , Organoides/citologia , Adulto , Células Cultivadas , Endométrio/crescimento & desenvolvimento , Endométrio/metabolismo , Feminino , Fertilização In Vitro , Humanos , Organoides/crescimento & desenvolvimento , Organoides/metabolismo , Projetos Piloto
9.
FASEB J ; 35(7): e21731, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-34131963

RESUMO

Successful embryo implantation requires well-functioning endometrial luminal epithelial cells to establish uterine receptivity. Inadequate uterine receptivity is responsible for approximately two thirds of implantation failures in humans. However, the regulatory mechanism governing this functional process remains largely unexplored. A previous study revealed that the expression of Rictor, the main member of mTORC2, in mouse epithelial cells is increased on the fourth day of gestation (D4). Here, we provide the first report of the involvement of Rictor in the regulation of endometrial receptivity. Rictor was conditionally ablated in the mouse endometrium using a progesterone receptor cre (PRcre ) mouse model. Loss of Rictor altered polarity remodeling and the Na+ channel protein of endometrial cells by mediating Rac-1/PAK1(pPAK1)/ERM(pERM) and Sgk1/pSgk1 signaling, respectively, ultimately resulting in impaired fertility. In the endometrium of women with infertility, the expression of Rictor was changed, along with the morphological transformation and Na+ channel protein of epithelial cells. Our findings demonstrate that Rictor is crucial for the establishment of uterine receptivity in both mice and humans. The present study may help improve the molecular regulatory network of endometrial receptivity and provide new diagnostic and treatment strategies for infertility.


Assuntos
Endométrio/metabolismo , Células Epiteliais/metabolismo , Alvo Mecanístico do Complexo 2 de Rapamicina/metabolismo , Proteína Companheira de mTOR Insensível à Rapamicina/metabolismo , Adulto , Animais , Modelos Animais de Doenças , Implantação do Embrião/fisiologia , Feminino , Fertilidade/fisiologia , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Progesterona/metabolismo , Transdução de Sinais/fisiologia , Útero/metabolismo , Adulto Jovem
10.
Nat Commun ; 12(1): 3386, 2021 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-34099644

RESUMO

During early pregnancy in the mouse, nidatory estrogen (E2) stimulates endometrial receptivity by activating a network of signaling pathways that is not yet fully characterized. Here, we report that bone morphogenetic proteins (BMPs) control endometrial receptivity via a conserved activin receptor type 2 A (ACVR2A) and SMAD1/5 signaling pathway. Mice were generated to contain single or double conditional deletion of SMAD1/5 and ACVR2A/ACVR2B receptors using progesterone receptor (PR)-cre. Female mice with SMAD1/5 deletion display endometrial defects that result in the development of cystic endometrial glands, a hyperproliferative endometrial epithelium during the window of implantation, and impaired apicobasal transformation that prevents embryo implantation and leads to infertility. Analysis of Acvr2a-PRcre and Acvr2b-PRcre pregnant mice determined that BMP signaling occurs via ACVR2A and that ACVR2B is dispensable during embryo implantation. Therefore, BMPs signal through a conserved endometrial ACVR2A/SMAD1/5 pathway that promotes endometrial receptivity during embryo implantation.


Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Implantação do Embrião , Infertilidade Feminina/genética , Receptores de Activinas Tipo II/genética , Receptores de Activinas Tipo II/metabolismo , Animais , Biópsia , Modelos Animais de Doenças , Endométrio/metabolismo , Endométrio/patologia , Estrogênios/metabolismo , Feminino , Humanos , Camundongos , Camundongos Knockout , Gravidez , Transdução de Sinais/fisiologia , Proteína Smad1/análise , Proteína Smad1/genética , Proteína Smad1/metabolismo , Proteína Smad5/análise , Proteína Smad5/genética , Proteína Smad5/metabolismo
11.
Int J Mol Sci ; 22(11)2021 May 26.
Artigo em Inglês | MEDLINE | ID: mdl-34073234

RESUMO

Embryonic implantation is a key step in the establishment of pregnancy. In the present work, we have carried out an in-depth proteomic analysis of the secretome (extracellular vesicles and soluble proteins) of two bovine blastocysts embryonic trophectoderm primary cultures (BBT), confirming different epithelial-mesenchymal transition stages in these cells. BBT-secretomes contain early pregnancy-related proteins and angiogenic proteins both as cargo in EVs and the soluble fraction. We have demonstrated the functional transfer of protein-containing secretome between embryonic trophectoderm and maternal MSC in vitro using two BBT primary cultures eight endometrial MSC (eMSC) and five peripheral blood MSC (pbMSC) lines. We observed that eMSC and pbMSC chemotax to both the soluble fraction and EVs of the BBT secretome. In addition, in a complementary direction, we found that the pattern of expression of implantation proteins in BBT-EVs changes depending on: (i) their epithelial-mesenchymal phenotype; (ii) as a result of the uptake of eMSC- or pbMSC-EV previously stimulated or not with embryonic signals (IFN-); (iii) because of the stimulation with the endometrial cytokines present in the uterine fluid in the peri-implantation period.


Assuntos
Quimiotaxia , Ectoderma/metabolismo , Implantação do Embrião , Embrião de Mamíferos/metabolismo , Endométrio/metabolismo , Células-Tronco Mesenquimais/metabolismo , Animais , Bovinos , Linhagem Celular , Feminino , Proteômica
12.
Int J Mol Sci ; 22(10)2021 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-34069423

RESUMO

Endometrosis is a reproductive pathology that is responsible for mare infertility. Our recent studies have focused on the involvement of neutrophil extracellular traps enzymes, such as elastase (ELA), in the development of equine endometrosis. Noscapine (NOSC) is an alkaloid derived from poppy opium with anticough, antistroke, anticancer, and antifibrotic properties. The present work investigates the putative inhibitory in vitro effect of NOSC on collagen type I alpha 2 chain (COL1A2) mRNA and COL1 protein relative abundance induced by ELA in endometrial explants of mares in the follicular or mid-luteal phases at 24 or 48 h of treatment. The COL1A2 mRNA was evaluated by qPCR and COL1 protein relative abundance by Western blot. In equine endometrial explants, ELA increased COL 1 expression, while NOSC inhibited it at both estrous cycle phases and treatment times. These findings contribute to the future development of new endometrosis treatment approaches. Noscapine could be a drug capable of preventing collagen synthesis in mare's endometrium and facilitate the therapeutic approach.


Assuntos
Colágeno Tipo I/metabolismo , Endometriose/metabolismo , Noscapina/farmacologia , Animais , Colágeno/metabolismo , Colágeno Tipo I/efeitos dos fármacos , Colágeno Tipo I/genética , Endometriose/tratamento farmacológico , Endometriose/veterinária , Endométrio/efeitos dos fármacos , Endométrio/metabolismo , Ciclo Estral , Armadilhas Extracelulares/metabolismo , Feminino , Fibrose , Doenças dos Cavalos/patologia , Cavalos , Noscapina/metabolismo , Elastase Pancreática/metabolismo , Inibidores de Proteases/farmacologia
13.
Ecotoxicol Environ Saf ; 220: 112361, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34052757

RESUMO

Exposure to ethephon (ETH), a plant growth regulator commonly used for several purposes, can potentially decrease sperm numbers and viability. Occasional findings regarding ETH effects on female reproduction during early pregnancy have also been reported. During early pregnancy, endometrial decidualization is a critical event for embryo implantation and pregnancy maintenance. Thus, we aimed to explore the effect and mechanism of ETH on endometrial decidualization both in vivo and in vitro. Mice were gavaged with 0 and 285 mg/kg b.w. ETH from gestational days (GD)1 until sacrifice, whereas pseudopregnant mice from pseudopregnant day 1 (PPD-1) until PPD-8. Primary mouse endometrial stromal cells (mESCs) received 640 ug/ml ETH and added E2 and P4 to induce decidualization. Results indicated female albino CD1 mice exposed to high dose of ETH (285 mg/kg b.w.) by oral gavage, the number of embryo implantation sites on GD6 and GD8 were significantly decreased, the levels of serum E2 and P4 on GD8 were significantly decreased. Compared with the control group, the decidualization response artificially induced by corn oil in pseudopregnant mice and by E2 and P4 in primary mouse endometrial stromal cells (mESCs) was weakened in the high dose of ETH treated group. The high dose, 285 mg/kg b.w ETH treated group altered the expression of endometrial decidual markers on GD6 and GD8. The triglyceride and fatty acid metabolism-related genes were significantly increased after female albino CD1 mice exposed to high does, 285 mg/kg b.w ETH on GD6 and GD8. GPR120 was substantially reduced after ETH treatment. When overexpression of GPR120, the compromised decidualization induced by ETH treatment was rescued. Furthermore, molecular docking presented Thr234 and His251 of GPR120 as preferred binding sites for ETH. Mutation of these two sites rescued the compromised decidualization induced by ETH. In conclusion, we demonstrated that ETH exposure could impair decidualization during early pregnancy. GPR120 expression and binding between GPR120 and ETH are crucial for impaired decidualization mediated via ETH.


Assuntos
Endométrio/efeitos dos fármacos , Compostos Organofosforados/toxicidade , Reguladores de Crescimento de Plantas/toxicidade , Receptores Acoplados a Proteínas G/metabolismo , Animais , Decídua/efeitos dos fármacos , Decídua/metabolismo , Decídua/patologia , Implantação do Embrião/efeitos dos fármacos , Endométrio/metabolismo , Endométrio/patologia , Feminino , Camundongos , Simulação de Acoplamento Molecular , Compostos Organofosforados/química , Reguladores de Crescimento de Plantas/química , Gravidez , Receptores Acoplados a Proteínas G/química , Células Estromais/efeitos dos fármacos , Células Estromais/metabolismo , Células Estromais/patologia
14.
Commun Biol ; 4(1): 572, 2021 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-33990675

RESUMO

Seminal fluid factors modulate the female immune response at conception to facilitate embryo implantation and reproductive success. Whether sperm affect this response has not been clear. We evaluated global gene expression by microarray in the mouse uterus after mating with intact or vasectomized males. Intact males induced greater changes in gene transcription, prominently affecting pro-inflammatory cytokine and immune regulatory genes, with TLR4 signaling identified as a top-ranked upstream driver. Recruitment of neutrophils and expansion of peripheral regulatory T cells were elevated by seminal fluid of intact males. In vitro, epididymal sperm induced IL6, CXCL2, and CSF3 in uterine epithelial cells of wild-type, but not Tlr4 null females. Collectively these experiments show that sperm assist in promoting female immune tolerance by eliciting uterine cytokine expression through TLR4-dependent signaling. The findings indicate a biological role for sperm beyond oocyte fertilization, in modulating immune mechanisms involved in female control of reproductive investment.


Assuntos
Implantação do Embrião/imunologia , Endométrio/imunologia , Tolerância Imunológica/imunologia , Reprodução , Espermatozoides/fisiologia , Linfócitos T Reguladores/imunologia , Útero/imunologia , Animais , Comunicação Celular , Citocinas/genética , Citocinas/metabolismo , Endométrio/metabolismo , Feminino , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Transdução de Sinais , Linfócitos T Reguladores/metabolismo , Útero/metabolismo , Vasectomia
15.
Biomed Res Int ; 2021: 6658321, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33937407

RESUMO

A thin endometrium affects the success of assisted reproduction due to low endometrial receptivity. Acupuncture improves endometrial receptivity and promotes the formation of pinopodes, the ultrastructure marker implantation window. However, the specific underlying mechanisms remain unclear. In this study, the efficacy of acupuncture treatment and its underlying mechanism were investigated by analyzing pregnancy rate, pinopode formation, and related molecular markers in thin endometrium model rats. Absolute ethanol (95%) was injected into the uteruses of female Sprague-Dawley rats to construct a thin endometrium model. In this model, acupuncture stimulation at EX-CA1, SP6, and CV4 ameliorated the pregnancy rate. Significantly increased embryo implantation, endometrial thickness, numbers of glands, and blood vessels were observed in the electroacupuncture (EA) group compared to the model group. The number of pinopodes in the EA group was abundant, with a shape similar to that of the control group. Additionally, significantly higher expression levels of pinopode-related markers, including integrin αvß3, homeobox A10 (HOXA10), heparin-binding EGF-like growth factor (HBEGF), estrogen receptor alpha (ERα), and progesterone receptor (PR), were observed in the EA group than those in the model group. In conclusion, EA had a positive effect on the endometrial receptivity of thin endometrium model rats by improving pinopode formation through multiple molecular targets.


Assuntos
Eletroacupuntura , Endométrio/metabolismo , Endométrio/patologia , Resultado da Gravidez , Animais , Modelos Animais de Doenças , Implantação do Embrião , Endométrio/ultraestrutura , Receptor alfa de Estrogênio/metabolismo , Ciclo Estral , Feminino , Masculino , Gravidez , Ratos Sprague-Dawley , Receptores de Progesterona/metabolismo
16.
PLoS One ; 16(4): e0249775, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33826645

RESUMO

BACKGROUND: The purpose of the present study was to evaluate the relationship between chronic endometritis and the epithelial-mesenchymal transition in the endometrium of infertile patients in the implantation phase. METHODS: Endometrial biopsy specimens from 66 infertility patients were analyzed. The presence of chronic endometritis was investigated by immunostaining for CD138. Immunohistochemical staining for E-cadherin, N-cadherin, Slug, and Snail was performed, and the expression profiles were statistically analyzed according to the presence of chronic endometritis. When the loss of E-cadherin expression and/or the positive expression of N-cadherin was detected, the specimen was considered epithelial-mesenchymal transition-positive. Epithelial-mesenchymal transition-positive cases were also statistically analyzed according to the presence of chronic endometritis. The characteristics of the patients in the epithelial-mesenchymal transition-positive and epithelial-mesenchymal transition-negative groups were compared. The association between variables, including age, body mass index, gravidity, parity, and each causative factor of infertility and epithelial-mesenchymal transition positivity was analyzed. RESULTS: The rates of the loss of E-cadherin expression, the gain of N-cadherin and epithelial-mesenchymal transition positivity were significantly higher in chronic endometritis patients. The expression of Slug, cytoplasmic Snail, and nuclear Snail was also detected at significantly higher rates in chronic endometritis patients. Chronic endometritis were related to the epithelial-mesenchymal transition. CONCLUSION: The epithelial-mesenchymal transition was frequently detected in the endometrium in infertile patients with chronic endometritis. Since the epithelial-mesenchymal transition is associated with chronic endometritis, the epithelial-mesenchymal transition appears to be involved in the alteration of mechanisms of implantation.


Assuntos
Endometrite/fisiopatologia , Endométrio/fisiologia , Transição Epitelial-Mesenquimal/fisiologia , Infertilidade/patologia , Adulto , Caderinas/metabolismo , Implantação do Embrião/fisiologia , Endometrite/metabolismo , Endométrio/metabolismo , Feminino , Humanos , Infertilidade/metabolismo , Fatores de Transcrição da Família Snail/metabolismo
17.
Int J Mol Sci ; 22(7)2021 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-33800594

RESUMO

Activation of trimethylation of histone 3 lysine 27 (H3K27me3) by EZH2, a component of the Polycomb repressive complex 2 (PRC2), is suggested to play a role in endometriosis. However, the mechanism by which this complex is dysregulated in endometriosis is not completely understood. Here, using eutopic and ectopic tissues, as well as peritoneal fluid (PF) from IRB-approved and consented patients with and without endometriosis, the expression of PRC2 complex components, JARID2, miR-155 (known regulators of EZH2), and a key inflammatory modulator, FOXP3, was measured. A higher expression of EZH2, H3K27me3, JARID2, and FOXP3 as well as miR-155 was noted in both the patient tissues and in endometrial PF treated cells. Gain-or-loss of function of miR-155 showed an effect on the PRC2 complex but had little effect on JARID2 expression, suggesting alternate pathways. Chromatin immunoprecipitation followed by qPCR showed differential expression of PRC2 complex proteins and its associated binding partners in JARID2 vs. EZH2 pull down assays. In particular, endometriotic PF treatment increased the expression of PHF19 (p = 0.0474), a gene silencer and co-factor that promotes PRC2 interaction with its targets. Thus, these studies have identified the potential novel crosstalk between miR-155-PRC2 complex-JARID2 and PHF19 in endometriosis, providing an opportunity to test other epigenetic targets in endometriosis.


Assuntos
Líquido Ascítico/metabolismo , Endometriose/metabolismo , Endométrio/metabolismo , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Regulação da Expressão Gênica , MicroRNAs/metabolismo , Adolescente , Adulto , Metilação de DNA , Feminino , Fatores de Transcrição Forkhead/metabolismo , Perfilação da Expressão Gênica , Humanos , Metilação , Pessoa de Meia-Idade , Peritônio/metabolismo , Complexo Repressor Polycomb 2/metabolismo , Regiões Promotoras Genéticas , Adulto Jovem
18.
Aging (Albany NY) ; 13(9): 12607-12630, 2021 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-33901012

RESUMO

Novel biomarkers are needed to accelerate the diagnosis and treatment of endometriosis. We performed RNA sequencing to explore the expression profiles of exosomal circular RNAs (circRNAs), microRNAs (miRNAs) and mRNAs in patients with ovarian endometriomas, eutopic endometria and normal endometria. Differentially expressed genes between the different pairs of groups were analyzed and functionally annotated. Then, miRNA-target RNA pairs were identified, competing endogenous RNA (ceRNA) scores were calculated, gene expression characteristics were determined, and these parameters were used to construct an exosomal ceRNA network. We identified 36 candidate hub genes with high degrees of gene connectivity. We also topologically analyzed the ceRNA network to obtain a hub ceRNA network of circRNAs with the highest closeness and ceRNA efficiency. Twelve genes overlapped between the 36 candidate hub genes and the genes in the hub ceRNA network. These 12 genes were considered to be exosomal RNA-based biomarkers, and circ_0026129/miRNA-15a-5p/ATPase H+ transporting V1 subunit A (ATP6V1A) were at the center of the ceRNA network. By determining the exosomal RNA expression profiles of endometriosis patients and constructing a circRNA-associated ceRNA network, these findings provide insight into the molecular pathways of endometriosis and new resources for its diagnosis and treatment.


Assuntos
Endometriose/metabolismo , Redes Reguladoras de Genes/genética , MicroRNAs/genética , RNA Circular/genética , Endometriose/genética , Endométrio/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Humanos , MicroRNAs/metabolismo , RNA Circular/metabolismo , RNA Mensageiro/metabolismo , Análise de Sequência de RNA/métodos
19.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 52(2): 235-240, 2021 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-33829697

RESUMO

Objective: To explore the influence for combination of Dingkun Dan with estradiol valerateon in treating rats with thin endometrium with Kidney-Yang deficiency based on Wnt/ß-catenin signaling pathway. Methods: The estrous period 40 rats were randomly divided into the normal control group, the model group, the Dingkun Dan group, the estradiol valerateon group and the combination group, with 8 rats in each group. In addition to the normal control group, the rat model of thin endometrium with Kidney-Yang deficiency was established in other groups. The control group used free diet, the model group was given distilled water, the estradiol valerateon group was treated with progynova by gavage at 0.3 mg/(kg·d) , Dingkun Dan group was treated with Dingkun Dan by gavage at 2.26 g/(kg·d), and the combined group was given Dingkun Dan at 2.26 g/(kg·d) on the basis of progynova at 0.3 mg/(kg·d). After 3 estrous cycles, the rats were killed and harvested. HE staining was used to observe histopathologic changes in endometrium. The expression of VEGF in rats endometrium were detected by immunohistochemistry. The expression of ß-catenin, E-cadherin and MMP-9 protein in rat endometrium was detected by Western blot. Results: Compared with the control group, the uterine cavity was narrowed or enlarged, the endometrium glands and blood vessels were sparse, and the endometrium was thinner significantly in the model group ( P<0.01); the levels of VEGF was decreased significantly ( P<0.01), while ß-catenin, E-cadherin and MMP-9 were increased significantly ( P<0.01). Compared with the model group, more endometrial glands, rich intimal vessels, the endometrium were thickened significantly in the 3 treatment groups ( P<0.01 or P<0.05); the levels of VEGF was increased differently. The protein levels of ß-catenin and E-cadherin were significantly decreased in each treatment group ( P<0.01), and MMP-9 were significantly decreased in the Dingkun Dan group and in the combination group ( P<0.01). Compared with the estradiol valerateon group, the level of ß-catenin in Dingkun Dan group was higher, and MMP-9 was lower ( P<0.05 or P<0.01). The levels of ß-catenin and MMP-9 in the combination group were significantly decreased ( P<0.01). Compared with the combination group, the levels of ß-catenin was increased significantly, while decreased signicantly in Dingkun Dan group ( P<0.01 or P<0.05). Conclusion: Dingkun Dan combined with estradiol valerateon can increase the thicken of the endometrium by up-regulation of VEGF, while down-regulate of ß-catenin, E-cadherin and MMP-9 in rats with Shen-Yang deficiency and thin endometrium.


Assuntos
Estradiol , Via de Sinalização Wnt , Animais , Endométrio/metabolismo , Feminino , Rim , Ratos , Deficiência da Energia Yang , beta Catenina/genética , beta Catenina/metabolismo
20.
Mol Med Rep ; 23(5)2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33760149

RESUMO

Endometriosis (EM) is a multifactorial and debilitating chronic benign gynecological disease, but the pathogenesis of the disease is not completely understood. Dysregulated expression of microRNAs (miRNA/miR) is associated with the etiology of EM due to their role in regulating endometrial stromal cell proliferation and invasion. The present study aimed to identify the functions and mechanisms underlying miR­143­3p in EM. To explore the role of miR­143­3p in EM, functional miRNAs were analyzed via bioinformatics analysis. miR­143­3p expression levels in endometriotic stromal cells (ESCs) and normal endometrial stromal cells (NESCs) were measured via reverse transcription­quantitative PCR. The role of miR­143­3p in regulating ESC proliferation and invasion was assessed by performing Cell Counting Kit­8 and Transwell assays, respectively. miR­143­3p expression was significantly upregulated in ESCs compared with NESCs. Functionally, miR­143­3p overexpression inhibited ESC proliferation and invasion, whereas miR­143­3p knockdown promoted ESC proliferation and invasion. Moreover, miR­143­3p inhibited autophagy activation in ESCs, as indicated by decreased green puncta, which represented autophagic vacuoles, decreased microtubule associated protein 1 light chain 3α expression and increased p62 expression in the miR­143­4p mimic group compared with the control group. Moreover, compared with the control group, miR­143­3p overexpression significantly decreased the expression levels of autophagy­related 2B (ATG2B), a newly identified target gene of miR­143­3p, in ESCs. ATG2B overexpression reversed miR­143­3p overexpression­mediated inhibition of ESC proliferation and invasion. Collectively, the results of the present study suggested that miR­143­3p inhibited EM progression, thus providing a novel target for the development of therapeutic agents against EM.


Assuntos
Proteínas Relacionadas à Autofagia/genética , Autofagia/genética , Endometriose/genética , MicroRNAs/genética , Proteínas de Transporte Vesicular/genética , Adulto , Movimento Celular/genética , Proliferação de Células/genética , Endometriose/patologia , Endométrio/metabolismo , Endométrio/patologia , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Regulação da Expressão Gênica/genética , Humanos , Pessoa de Meia-Idade , Células Estromais/metabolismo , Células Estromais/patologia
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