The aim of the present study was to investigate the underlying mechanisms of hypoxia-induced microRNA (miR)-210 effects on mouse GC-2spd (GC-2) cells. GC-2 cells were subjected to hypoxia or normoxia for 12, 24, 48 and 72 h. Apoptosis of GC-2 cells was detected using terminal deoxynucleotidyl-transferase-meditated dUTP nick end labeling and flow cytometry. Reverse transcription-quantitative polymerase chain reaction was performed to analyze the expression of miR-210. Hypoxia-inducible factor-1α (HIF-1α), caspase-3, B-cell lymphoma 2, apoptosis regulator BAX and Kruppel-like factor 7 (KLF7) protein expression levels were detected by western blotting. Luciferase reporter gene assays were used to assess the targeting effects of miR-210 on KLF7. Hypoxia induced GC-2 cell apoptosis and increased the expression of HIF-1α and pro-apoptotic proteins; however, decreased anti-apoptotic protein expression levels. Furthermore, hypoxia resulted in the upregulation of miR-210 in GC-2 cells. HIF-1α and miR-210 were involved in the apoptosis of GC-2 cells by mediating the expression of apoptosis-associated proteins. Furthermore, KLF7 was directly targeted by miR-210 to influence the apoptosis of GC-2 cells subjected to hypoxia. The results suggested that hypoxia-induced miR-210 stimulated the activation of the apoptosis signaling pathway and contributed to the apoptosis of GC-2 cells by targeting KLF7.
Background: Repeated implantation failure (RIF) is currently believed by some scholars to be mainly related to the poor endometrial receptivity. Three-dimensional (3D) ultrasound, as a noninvasive examination, has attracted the most attention. This paper further discusses whether 3D ultrasound of the endometrial receptivity index has guidance value in the evaluation of pregnancy outcomes in patients with RIF following frozen-thawed embryo transfer.Methods: A total of 79 patients with RIF were retrospectively analyzed. These 79 patients which were confirmed by the transvaginal ultrasonography detection comprised 36 cases of intrauterine pregnancy, which were included in the pregnancy group, and 43 cases with biochemical pregnancy (the HCG in blood or urine is only transient and can be detected by clinical biochemical methods. The pregnancy sac cannot be seen under ultrasound) or negative results of human chorionic gonadotropin (HCG) examination, which were included in the non-pregnancy group. The endometrial thickness, uterine volume, sub-endometrial blood flow type, pulsatility index (PI) and resistance index (RI) of the spiral artery; the RI and PI of the uterine artery; and the peak systolic velocity/end diastolic velocity (S/D) of the two groups were measured and analyzed.Results: (I) There were no significant differences in the age, infertility years, body mass index (BMI), anti-Müllerian hormone (AMH), endometrial thickness on the hCG injection day, estradiol (E2), and progesterone (P) between the pregnant and non-pregnant groups (P>0.05). (II) There were no significant differences in the endometrial thickness, uterine volume, and RI and PI of the uterine artery on the day before the implantation between the two groups (P>0.05). (III) There was statistical difference in the sum of the S/D of the two uterine arteries between the two groups (P<0.05) with a cutoff value of 14.47 (P<0.05).(IV) The RI and PI of the spiral artery in the non-pregnancy group were lower than those in the pregnancy group, and the difference was statistically significant (P<0.05). (V) The differences in the endometrial classification and subendometrial blood flow classification between the two groups were statistically significant (P<0.05).Conclusions: Ultrasonographic endometrial blood flow classification, spiral artery blood flow parameters, and uterine artery blood flow parameters can be effective indexes for evaluating endometrial receptivity, and they have a certain clinical significance in evaluating the pregnancy outcome of RIF patients after retransplantation.
The aim of this study was to investigate the underlying mechanisms of hypoxia-induced apoptosis of GC-2spd (GC-2) cells. The GC-2 cells were treated with or without hypoxia for 12, 24, 48, and 72 h. Apoptosis of GC-2 cells was detected using TUNEL and flow cytometry. Fluorescence microscopic was used to observe the autophagy of GC-2 cells. Hypoxia-inducible factor-1alpha (HIF-1α), apoptosis-related protein and marker protein of autophagy levels were measured by Western blotting. Hypoxia induced apoptosis and autophagy of GC-2 cells, and increased expression of HIF-1α, LC3-II, Beclin-1, and pro-apoptotic protein, but reduced p62 and anti-apoptotic protein level. Meanwhile, hypoxia-induced HIF-1α mediated expression of apoptosis and autophagy-related protein in GC-2 cell. Furthermore, autophagy regulated hypoxia-induced apoptosis of GC-2 cell. Our data suggest that hypoxia induces apoptosis of GC-2 cell by activation of autophagy involving activation of the apoptosis signaling pathway under the hypoxic condition.
The normal development and maturation of oocytes and sperm, the formation of fertilized ova, the implantation of early embryos, and the growth and development of foetuses are the biological basis of mammalian reproduction. Therefore, research on oocytes has always occupied a very important position in the life sciences and reproductive medicine fields. Various embryo engineering technologies for oocytes, early embryo formation and subsequent developmental stages and different target sites, such as gene editing, intracytoplasmic sperm injection (ICSI), preimplantation genetic diagnosis (PGD), and somatic cell nuclear transfer (SCNT) technologies, have all been established and widely used in industrialization. However, as research continues to deepen and target species become more advanced, embryo engineering technology has also been developing in a more complex and sophisticated direction. At the same time, the success rate also shows a declining trend, resulting in an extension of the research and development cycle and rising costs. By studying the existing embryo engineering technology process, we discovered three critical nodes that have the greatest impact on the development of oocytes and early embryos, namely, oocyte micromanipulation, oocyte electrical activation/reconstructed embryo electrofusion, and the in vitro culture of early embryos. This article mainly demonstrates the efforts made by researchers in the relevant technologies of these three critical nodes from an engineering perspective, analyses the shortcomings of the current technology, and proposes a plan and prospects for the development of embryo engineering technology in the future.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.