RESUMEN
This study investigates the influence of four culture media pre-equilibration methods on embryo development and clinical pregnancy outcomes. The methods are as follows: Method A involved covering media with fresh mineral oil in humid-type incubators for 24 h. Method B replicated Method A in dry-type incubators. Method C utilized pre-equilibrated (humidified) mineral oil to cover the media, also in humid-type incubators for 24 h. Method D followed the same process as Method C but in dry-type incubators. Subsequently, media from all groups were transferred to dry-type incubators for 72 h. Osmolality was measured at 24, 48, 72, and 96 h. For G1 PLUS, no significant differences were observed among groups at 24, 48, and 72 h. However, at 96 h, Groups B and D exhibited significantly higher osmolality than Groups A and C (A vs B, p = 0.043; A vs D: p = 0.046; B vs C, p = 0.043; C vs D, p = 0.046). No significant variations were found between Groups A and C or B and D. Similar results were obtained for G2 PLUS. A retrospective analysis of embryo development and clinical outcomes using Methods A revealed significant improvements in good blastocysts and available embryos compared with Method B for all (p = 0.005 and 0.004) and IVF cycles (p = 0.025 and 0.017). Method A also enhanced blastocyst formation in ICSI cycles (p = 0.017). However, clinical pregnancy outcomes did not significantly differ between Methods A and B. Pre-equilibrating culture media overnight in humid-type incubators, even when covered with fresh mineral oil, significantly mitigates osmolality rise and improves embryo development potential during embryo culture in dry-type incubators.
Asunto(s)
Medios de Cultivo , Técnicas de Cultivo de Embriones , Desarrollo Embrionario , Medios de Cultivo/química , Desarrollo Embrionario/fisiología , Femenino , Técnicas de Cultivo de Embriones/métodos , Embarazo , Humanos , Adulto , Concentración Osmolar , Fertilización In Vitro/métodos , Estudios Retrospectivos , Transferencia de Embrión/métodosRESUMEN
Decidualization is a progesterone-dependent cellular differentiation process that is essential for establishing pregnancy. Robust activation of glycolysis and lactate synthesis during decidualization is remarkable, but their developmental functions remain largely unknown. Herein, we identify that endometrial lactate production plays a critical role in establishing local histone lactylation, a newly identified histone modification, and is important for ensuring normal decidualization. Enhanced endometrial glycolysis is the hallmark metabolic change and is tightly coupled with H4K12la during decidualization. Inhibition of histone lactylation impaired decidualization, in either physiological conception or in vivo and in vitro induced decidualization models. Mechanistic study based on CUT&Tag and ATAC-seq revealed that a transcriptional factor hypoxia-inducible factor 1 α (Hif1α) is the critical regulatory target of H4K12la, and in turn forms an H4K12la-Hif1α-glycolysis feedback loop to drive decidualization. Moreover, we demonstrate that the loop is directly activated by progesterone during decidualization. Our study not only advances the current knowledge of the role of lactate in regulating uterine function, but also establishes a novel functional link among the major endocrine factors, endometrial metabolic change, and epigenetic modification during endometrial remodeling. These findings present valuable clues to develop clinical intervention strategies to improve pregnancy outcomes following both natural conception and assisted reproduction.
Asunto(s)
Histonas , Progesterona , Embarazo , Femenino , Humanos , Progesterona/farmacología , Progesterona/metabolismo , Histonas/metabolismo , Decidua/metabolismo , Retroalimentación , Endometrio/metabolismo , Lactatos/metabolismo , Glucólisis , Células del Estroma/metabolismoRESUMEN
A fixed-frequency beam-scanning leaky-wave antenna (LWA) array with three switchable dual-polarized beams is proposed and experimentally demonstrated. The proposed LWA array consists of three groups of spoof surface plasmon polaritons (SPPs) LWAs with different modulation period lengths and a control circuit. Each group of SPPs LWAs can independently control the beam steering at a fixed frequency by loading varactor diodes. The proposed antenna can be configured in both multi-beam mode and single-beam mode, where the multi-beam mode with optional two or three dual-polarized beams. The beam width can be flexibly adjusted from narrow to wide by switching between multi-beam and single-beam states. The prototype of the proposed LWA array is fabricated and measured, and both simulation and experimental results show that the antenna can accomplish a fixed frequency beam scanning at an operating frequency of 3.3 to 3.8 GHz with a maximum scanning range of about 35° in multi-beam mode and about 55° in single-beam mode. It could be a promising candidate for application in the space-air-ground integrated network scenario in satellite communication and future 6G communication systems.
Asunto(s)
Comunicaciones por Satélite , Simulación por Computador , CintigrafíaRESUMEN
The aim of the study was to examine the clinical value of blastocysts derived from mono-pronuclear (1PN) or non-pronuclear (0PN) zygotes with two polar bodies (2PB), which were selected by our criteria. We retrospectively analysed 610 frozen-thawed blastocyst transfer (FET) cycles and the corresponding oocyte retrieval cycles from 2014 to 2017. Developmental potential and clinical outcomes of embryos derived from zygotes with various numbers of pronuclei were analysed. Based on more detailed pre-selection settings, blastulation rates of 1PN/2PB and 0PN/2PB-derived embryos were 70.18% and 69.17%, respectively. Blastocyst FET results were not significantly different between 2PN/2PB, 1PN/2PB and 0PN/2PB groups in terms of clinical pregnancy rates (59.79%, 47.06% and 56.25%), implantation rates (47.24%, 40.00% and 47.62%), live birth rates (49.39%, 29.41% and 43.75%) or malformation rates (0%, 0% and 0%). In conclusion, after strict morphological selection and blastocyst culture, 1PN/2PB and 0PN/2PB-derived embryos in IVF cycles can have considerable clinical value. Blastocysts derived from 1PN/2PB or 0PN/2PB zygotes are worthwhile FET option for patients who have no available 2PN-derived embryos.
Asunto(s)
Blastocisto , Fertilización In Vitro , Transferencia de Embrión/métodos , Femenino , Fertilización In Vitro/métodos , Humanos , Embarazo , Índice de Embarazo , Estudios RetrospectivosRESUMEN
The purpose of this study was to explore the application of glucose to improve embryo development potential when fertilization is delayed in mice. After recovery, mouse oocytes were cultured alone for 6 h before fertilization in three fertilization media: G-IVF PLUS, G-IVF PLUS with 5 mM and 10 mM glucose. G-IVF PLUS group was used as the control group. Then, in vitro fertilization (IVF) was performed and blastocysts were transferred at Day 4. To compare the effects of different glucose concentrations on embryo development and birth outcomes, conventional IVF and embryo transfer were carried out in G-IVF PLUS, G-IVF PLUS with 5 mM and 10 mM glucose. The results indicated that G-IVF PLUS with 5 mM glucose significantly increased blastocyst rate (p < 0.05) and birth rate (p < 0.05) when fertilization was delayed 6 h compared with G-IVF PLUS groups. In conventional IVF without delayed fertilization, embryo development was not significantly affected by G-IVF PLUS with 5 mM or 10 mM glucose. There were no significant differences in terms of birth rate, fetal weight, crown-rump length, tail length and birth defect rate among the three groups. In conclusion, 5 mM glucose could significantly improve embryo developmental potential and birth outcomes when fertilization was delayed 6 h and did not have adverse effects on embryo quality and birth outcomes for normal IVF. It might have a good prospect of clinical application in assisted reproductive technology (ART). Abbreviations: ART: assisted reproductive technology; IVF: in vitro fertilization; ICSI: intracytoplasmic sperm injection; TFF: total fertilization failure; TESA: testicular sperm aspiration.
Asunto(s)
Blastocisto/efectos de los fármacos , Técnicas de Cultivo de Embriones , Fertilización In Vitro , Glucosa/farmacología , Oocitos/efectos de los fármacos , Animales , Blastocisto/fisiología , Relación Dosis-Respuesta a Droga , Implantación del Embrión , Transferencia de Embrión , Desarrollo Embrionario/efectos de los fármacos , Femenino , Nacimiento Vivo , Ratones Endogámicos ICR , Oocitos/fisiología , Embarazo , Índice de Embarazo , Factores de TiempoRESUMEN
The safety of assisted reproductive technology (ART) is of frequent concern. Unfortunately, animal models for studying the safety of intracytoplasmic sperm injection (ICSI) have limitations in mimicking human ICSI manipulations. As reported herein, we invented a modified holding pipette for mouse oocyte injection that resulted in the delivery of live pups. A modified holding pipette was prepared for mouse oocyte injection and was compared with the conventional pipette for human use and a trumpet-shaped pipette. After ICSI, the oocytes were cultured to cleavage embryos until fallopian transfer. The use of the trumpet-shaped holding pipette and the new modified holding pipette for mouse oocyte injection achieved comparable and satisfactory oocyte survival rates (83.44% and 85.71%, respectively) and embryo cleavage rates (41.98% and 42.42%, respectively), which were significantly higher than those obtained with the human egg-holding pipette (oocyte survival rate: 65.85%; embryo cleavage rate: 27.78%). After 13 embryos were transferred using each type of pipette, three live pups were produced with the new modified holding pipette, one was produced with the holding pipette for human use, and none were produced with the trumpet-shaped holding pipette. The modified holding pipette for oocyte injection is effective and very easy to prepare. Moreover, using this new method, we produced live pups, which will contribute to a useful animal model for safety studies of ICSI in the future.
Asunto(s)
Técnicas de Cultivo de Embriones/veterinaria , Inyecciones de Esperma Intracitoplasmáticas/veterinaria , Animales , Proteínas de Escherichia coli , Femenino , Proteínas de Transporte de Membrana , Ratones , Embarazo , Inyecciones de Esperma Intracitoplasmáticas/instrumentaciónRESUMEN
This study aimed to explore the effects of low-concentration (0.6 ng/mL) granulocyte-macrophage colony-stimulating factor (GM-CSF) supplementation on human embryo quality and pregnancy outcomes in patients with fresh transfer cycles. The data were retrospectively collected from 719 hyperstimulation cycles of 631 patients divided into two groups: GM-CSF supplementation (0.6 ng/mL, n = 399) and control (n = 320). The embryo quality and pregnancy outcomes were compared. GM-CSF addition significantly increased the available embryo rate (52.0 vs. 48.1%, p < .05). In patients >38 years, it significantly enhanced cleavage (99.4 vs. 97.8%, p < .05) and blastocyst formation (45.7 vs. 34.9%, p < .05) rates but not pregnancy outcomes, including clinical pregnancy (power = 0.160) and implantation (power = 0.204) rates. The lack of a statistically significant difference could be related to low study power. These results suggest that low-concentration GM-CSF addition contributes to embryo quality improvement, especially in patients >38 years.IMPACT STATEMENTWhat is already known on this subject? Granulocyte-macrophage colony-stimulating factor (GM-CSF) has a beneficial effect on the development of human embryos in assisted reproductive technology.What do the results of this study add? Adding 0.6 ng/mL GM-CSF significantly increased the available embryo rate. In patients over 38 years of age, it statistically significantly enhanced the cleavage rate (99.4 vs. 97.8%, p < .05) and blastocyst formation rate (45.7 vs. 34.9%, p < .05).What are the implications of these findings for clinical practice and/or further research? GM-CSF benefits embryos with poorer development potential and a randomised clinical trial with a larger sample size should be performed.
Asunto(s)
Medios de Cultivo/farmacología , Implantación del Embrión/efectos de los fármacos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Adulto , Factores de Edad , Estudios de Casos y Controles , Técnicas de Cultivo de Embriones/métodos , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/sangre , Humanos , Embarazo , Resultado del Embarazo , Estudios RetrospectivosRESUMEN
The purpose of this study was to compare the efficacy of different open cryo-carriers: the CryoloopTM, CryotopTM, and CryoleafTM, in embryo survival and clinical outcome in patients with frozen embryo transfer (FET) cycle. We analyzed the embryo survival rate and clinical outcome in 325 patients of 348 FET cycles vitrified with the CryoloopTM (160 cycles), CryotopTM (105 cycles), or CryoleafTM (83 cycles). No significant differences were observed in embryo survival rate (98.8% vs. 100% vs. 97.7%, p > 0.05), HCG positive rate (58.8% vs. 63.8% vs. 57.8%, p > 0.05), biochemical pregnancy rate (6.9% vs. 11.4% vs. 9.6%, p > 0.05), or implantation rate (33.2% vs. 37.4% vs. 34.1%, p > 0.05) in the three groups respectively. The early abortion rate of the CryoloopTM group was significantly higher than that of the CryotopTM and CryoleafTM group (27.1% vs. 3.6% and 7.5%, p < 0.05). At the same time, the average female age of the CryoloopTM group was significantly older by 1 year than that of the CryotopTM and CryoleafTM group (33.29 ± 4.71 years vs. 31.96 ± 4.27 years and 31.1 ± 4.28 years, p < 0.05). There was no significant difference in take home baby rate (38.1% vs. 46.7% vs. 43.4, p > 0.05) or birth weight among the groups (2893.5 ± 780.8 g vs. 2778.4 ± 710.0 g vs. 2724.5 ± 838.8 g, p > 0.05). No case of neonatal malformation was observed in the present study. Overall, CryotopTM and CryoleafTM were effective for embryo vitrification at both the cleavage and blastocyst stage according to the results of clinical outcome and infant characteristics. However, CryoloopTM led to a decreased positive HCG rate and increased early abortion rate, heightened at the cleavage stage. ABBREVIATIONS: LN2: liquid nitrogen; CPA: cryoprotectant; ART: assisted reproductive technology; IVF: in vitro fertilization; ICSI: intracytoplasmic sperm injection; BMI: body mass index; FSH: follicular stimulation hormone; COH: controlled ovarian hyperstimulation; FET: frozen embryo transfer; mm: millimeter; HCG: human chorionic gonadotropin; RCT: randomized clinical trial; NC: natural cycle; AC: artificial cycle; EM: equilibration medium; DMSO: dimethyl sulphoxide; EG: ethylene glycol; VM: vitrification medium; WM: warming medium.
Asunto(s)
Blastocisto/fisiología , Criopreservación/instrumentación , Transferencia de Embrión , Fertilización In Vitro , Infertilidad/terapia , Aborto Espontáneo/etiología , Adulto , Supervivencia Celular , Implantación del Embrión , Transferencia de Embrión/efectos adversos , Diseño de Equipo , Femenino , Fertilidad , Fertilización In Vitro/efectos adversos , Humanos , Infertilidad/diagnóstico , Infertilidad/fisiopatología , Nacimiento Vivo , Masculino , Edad Materna , Embarazo , Índice de Embarazo , Estudios Retrospectivos , Factores de Riesgo , Resultado del Tratamiento , VitrificaciónRESUMEN
Development of an effective system for oocyte-cryopreservation is of clinical relevance in reproductive medicine. However, oocyte-preservation is not as effective as embryo preservation. In this study, we used a 37°C pre-equilibrium temperature as part of a modified vitrification method for human oocyte cryopreservation. The effect of the new method on spindle configuration, chromosomal arrangement, and mitochondrial distribution was investigated in in vitro-matured human oocytes. A total of 101 in vitro-matured oocytes were randomly assigned for vitrification at pre-equilibrium temperature of 37°C (37°C Group, n=50) or at room temperature (RT Group, 22-24°C, n=51). The time needed for vitrification in the 37°C group was significantly shorter than that in the RT group. Defective spindles were found in 45.5% and 69.0% oocytes in the 37°C group and RT group, respectively (p < 0.05). Abnormal chromosomes were found in 47.7% and 71.4% oocytes, respectively (p < 0.05). There were no significant differences with respect to oocyte survival rate and mitochondrial distribution pattern between the two groups. These results indicate that vitrification at a pre-equilibrium temperature of 37°C may reduce the incidence of defective spindle configuration and chromosomal abnormalities in in-vitro-matured human oocytes. ABBREVIATIONS: ICSI: intracytoplasmic sperm injection; FSH: follicle-stimulating hormone; MII: metaphase II; EG: ethylene glycol; PROH: 1,2-propanediol.
Asunto(s)
Aberraciones Cromosómicas , Criopreservación/métodos , Oocitos , Vitrificación , Adulto , Femenino , HumanosRESUMEN
AIM: The study aimed to assess the effect of in vitro vitrification and maturation on the nuclear configuration, cytoplasmic maturation and global DNA methylation pattern of human germinal vesicle (GV) stage oocytes. METHODS: Human oocytes from infertile women were randomly assigned to one of 3 groups: (i) metaphase II (MII) oocytes matured in vivo (vivo-MII, n = 56) as controls; (ii) MII oocytes matured in vitro (vitro-MII, n = 106); and (iii) MII oocytes that were vitrified at the GV stage, warmed and matured in vitro (cryo-MII, n = 122). All MII oocytes were fixed and immunofluorescence staining for spindle, chromosome, mitochondrion and cortical granules (CGs) were performed; examination was done using immunofluorescence laser scanning confocal microscope. The expression of 5-methycytosine in these MII oocytes was also assessed. RESULTS: No significant difference was observed between vitro-MII and cryo-MII groups with respect to oocyte maturation rate (72.4 vs. 78.3%). No significant differences were observed in the distribution of mitochondria, migration of CG and global DNA methylation pattern among the 3 study groups. However, the abnormal configuration of spindle and chromosome was significantly higher in cryo-MII group (78.9 and 84.2%) as compared to that in the vitro-MII (45.0 and 50.0%, p < 0.05) and vivo-MII groups (27.3 and 36.4%, p < 0.05). CONCLUSION: GV oocytes appeared to resist vitrification and retain their potential for maturation to MII stage oocytes after thawing. However, GV oocytes vitrification combined with in vitro maturation could affect the organization of spindle and chromosome.
Asunto(s)
Criopreservación/métodos , Metilación de ADN , Oocitos/química , Oocitos/ultraestructura , Núcleo Celular , Femenino , Calor , Humanos , Técnicas de Maduración In Vitro de los Oocitos , Infertilidad Femenina , MetafaseRESUMEN
PURPOSE: The purpose of this study is to explore whether a low concentration of granulocyte-macrophage colony-stimulating factor (GM-CSF) supplementation in culture medium is beneficial to infertile women aged over 35 years. METHODS: A retrospective cohort study was performed to analyze the embryo quality and pregnancy outcome of 212 controlled ovarian stimulation (COH) cycles with or without GM-CSF addition (n = 117 [GM-CSF, 0.2 ng/mL] vs n = 95 [control]). RESULTS: No significant difference was observed in cleavage rate (96.2 vs 96.5 %), blastocyst formation rate (53.2 vs 54.0 %), good blastocyst rate (26.8 vs 26.8 %), or available embryo rate (54.2 vs 49.7 %) between the GM-CSF group and the control group. However, the average age of the GM-CSF group (38.41 ± 3.13 years) was significantly 1 year older than that of the corresponding control group (37.45 ± 2.74 years) (P < 0.05). GM-CSF addition greatly decreased the occurrence of biochemical pregnancy (55.6 % [control] vs 20.8 % [GM-CSF], P < 0.05). No case of neonatal malformation was observed in the present study. CONCLUSION: Although no benefit of GM-CSF on embryo quality was observed, the addition of this factor significantly decreased the occurrence of chemical pregnancy of women aged over 35 years, indicating the role of GM-CSF in improving implantation competence of embryos derived from elderly infertile women.
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Medios de Cultivo/farmacología , Técnicas de Cultivo de Embriones/métodos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Infertilidad Femenina/tratamiento farmacológico , Adulto , Blastocisto/efectos de los fármacos , Implantación del Embrión/efectos de los fármacos , Femenino , Fertilización In Vitro/métodos , Humanos , Infertilidad Femenina/patología , Oocitos/efectos de los fármacos , Oocitos/crecimiento & desarrollo , Inducción de la Ovulación , Embarazo , Resultado del Embarazo , Inyecciones de Esperma Intracitoplasmáticas/métodosRESUMEN
The aim of this study was to explore whether the morphology of polar bodies (PBs) estimated at 16-18 h after insemination can be used as an additional marker for predicting human embryo quality or pregnancy outcome. The data from 355 patients who received standard in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatment after controlled ovarian hyperstimulation were recruited. Normal fertilized 3048 zygotes from 382 cycles were divided into two groups, PBs intact or fragmented, according to the morphology of PBs assessed at 16-18 h after insemination. Embryo quality and pregnancy outcome were compared between the two groups. It was shown that the day 3 (D3) good embryo rate, good quality blastocyst rate and available embryo rate of the PBs intact group were all significantly higher than that of the corresponding fragmented groups. However, no significant differences in pregnancy rate (PR) or implantation rate (IR) were observed between the intact and fragmented groups. Although PBs morphology estimated at 16-18 h after insemination had little effect on PR or IR in fresh embryo transfer cycles, a better embryo quality can be achieved in the PB-intact group, which is valuable for embryo selection.
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Embrión de Mamíferos/metabolismo , Fertilización In Vitro/métodos , Cuerpos Polares/metabolismo , Resultado del Embarazo , Inyecciones de Esperma Intracitoplasmáticas/métodos , Adulto , Blastocisto/citología , Blastocisto/metabolismo , Blastómeros/citología , Blastómeros/metabolismo , Recuento de Células , Embrión de Mamíferos/citología , Femenino , Humanos , Masculino , Embarazo , Índice de EmbarazoRESUMEN
MiR-185 expression has been associated with many cancers. However, the roles of miR-185 in human breast cancer remain elusive. Here, we found that miR-185 expression was decreased in human breast cancer tissues compared with healthy tissue controls. Up-regulation of miR-185 inhibited breast cancer cell proliferation and invasion and vice versa. MiR-185 was shown to bind to the 3'-untranslated region (UTR) of vascular endothelial growth factor a (Vegfa), and a significant inverse correlation was found between miR-185 and Vegfa. Vegfa overexpression partially restored the inhibition of cell proliferation and invasion that was induced by miR-185, and vice versa. Additionally, Vegfa expression was found to be high in human breast cancer tissues. Thus, miR-185-mediated Vegfa targeting may be involved in breast cancer formation.
Asunto(s)
Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Carcinogénesis/genética , MicroARNs/genética , Factor A de Crecimiento Endotelial Vascular/genética , Apoptosis/genética , Biomarcadores de Tumor/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular , Supervivencia Celular/genética , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Invasividad Neoplásica , Metástasis de la NeoplasiaRESUMEN
Dibutyl phthalate (DBP), a widely used phthalate, is known to cause many serious diseases, especially in the reproductive system. However, little is known about the effects of its metabolite, mono-n-butyl phthalate (MBP), on preimplantation embryo development. In the present study, we found that treatment of embryos with 10⻳ M MBP impaired developmental competency, whereas exposure to 10â»4 M MBP delayed the progression of preimplantation embryos to the blastocyst stage. Furthermore, reactive oxygen species (ROS) levels in embryos were significantly increased following treatment with 10⻳ M MBP. In addition, 10⻳ M MBP increased apoptosis via the release of cytochrome c, whereas immunofluorescent analysis revealed that exposure of preimplantation embryos to MBP concentration-dependently (10â»5, 10â»4 and 10⻳ M) decreased DNA methylation. Together, the results indicate a possible relationship between MBP exposure and developmental failure in preimplantation embryos.
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Apoptosis/efectos de los fármacos , Blastocisto/efectos de los fármacos , Ectogénesis/efectos de los fármacos , Disruptores Endocrinos/toxicidad , Estrés Oxidativo/efectos de los fármacos , Ácidos Ftálicos/toxicidad , Solventes/toxicidad , Animales , Blastocisto/citología , Blastocisto/metabolismo , Blastómeros/citología , Blastómeros/efectos de los fármacos , Blastómeros/metabolismo , Fase de Segmentación del Huevo/citología , Fase de Segmentación del Huevo/efectos de los fármacos , Fase de Segmentación del Huevo/metabolismo , Metilación de ADN/efectos de los fármacos , Técnicas de Cultivo de Embriones , Femenino , Fertilización In Vitro , Masculino , Ratones Endogámicos ICR , Mórula/citología , Mórula/efectos de los fármacos , Mórula/metabolismo , Concentración Osmolar , Plastificantes/toxicidad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Mono (2-ethylhexyl) phthalate (MEHP), an environmental contaminant, is known to cause many serious diseases, especially in reproductive system. However, little is known about the effect of MEHP on preimplantation embryo development. In this study, we found that the development of mouse 2-cell embryo was blocked by 10(-3) M MEHP. A significant increase in the level of reactive oxygen species (ROS) was observed in arrested 2-cell embryo following 10(-3) M MEHP treatment for 24 h. However, antioxidants, catalase (CAT), and superoxide dismutase (SOD), reduced intracellular ROS and protected MEHP-exposed embryos from death but failed to return the arrested embryos. Further experiments demonstrated that the level of apoptosis was not altered in live arrested 2-cell embryo and increased in dead arrested 2-cell embryo after MEHP treatment, which implied that ROS and apoptosis were not related with 2-cell block. During analysis of the indicators of embryonic genome activation (EGA) initiation (Hsc70, MuERV-L, Hsp70.1, eIF-1A, and Zscan4) and maternal-effect genes (OCT4 and SOX2), we found that MEHP treatment could significantly decline Hsc70, MuERV-L mRNA level and SOX2 protein level, and markedly enhance Hsp70.1, eIF-1A, Zscan4 mRNA level, and OCT4 protein level at 2-cell to 4-cell stage. Supplementation of CAT and SOD did not reverse the expression tendency of EGA related genes. Collectively, this study demonstrates for the first time that MEHP-induced 2-cell block is mediated by the failure of EGA onset and maternal-effect genes, not oxidative stress and apoptosis.