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A newly designed tetracarboxylic acid ligand precursor 5,5'-([9,9'-bianthracene]-10,10'-diyl)diisophthalic acid (H4BADI) has been used to prepare a series of copper-based metal-organic frameworks (MOFs) with the formula [Cu2(BADI)(S)2]·xS (denoted as 1-S, where S = solvent) and exhibiting solvent-induced structural transformations. Single-crystal-to-single-crystal transformation occurs upon exchanging 1-DMF (DMF = N,N-dimethylformamide) with DMSO (DMSO = dimethylsulfoxide). 1-DMF exhibits reversible structural transformation upon treatment with a variety of solvents; of particular interest is the reversible crystalline-to-amorphous phase transformations observed upon exchange with volatile, polar solvents. A thorough structural investigation of the three framework isomers characterized via single-crystal X-ray diffraction experiments is reported and compared to several other tetracarboxylate-based MOFs composed of dimetal secondary building units.
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AIM OF THE STUDY: To study the cross-border regulation of immunity and energy metabolism by ginseng miRNA156, and to provide a new perspective for further exploring the possibility of ginseng miRNA156 as a pharmacodynamic substance. MATERIALS AND METHODS: Combined with the previous research results of our research group, miRNA156 with high expression in blood sequencing of intragastrically administered with ginseng decoction was selected. Bioinformatics analysis was performed on the selected differential miRNA156. The target genes of differential miRNA156 were mainly enriched in metabolic, immune and other signaling pathways. According to the analysis results, the experimental part will use qi deficiency fatigue model and RAW264.7 cells. The contents of lactic acid (LA), creatine kinase (CK), blood urea nitrogen (BUN), lactate dehydrogenase (LD), liver glycogen (LG), muscle glycogen (MG), interleukin 4 (IL-4), matrix metallo-proteinase 9 (MMP-9), superoxide dismutase (SOD), malondialdehyde, phosphor-enolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase (G6pase), nitric oxide (NO) and tumor necrosis factor-α (TNF-α) were measured after administration of miRNA156. RESULTS: Ginseng miRNA156 can accelerate the removal of metabolic waste during exercise. Increase the glycogen reserve in, provide energy for the body, regulate the activity of key gluconeogenesis enzyme phosphorus, improve the energy metabolism system of, and enhance the endurance of fatigue mice. The contents of matrix metalloproteinase 9, superoxide dismutase and malondialdehyde were affected, and the content of TNF-α in the supernatant of RAW264.7 cells was significantly increased, which had certain antioxidant capacity and potential immunomodulatory effects. CONCLUSION: Ginseng miRNA156 has a certain regulatory effect on the energy metabolism and immune function of mice, which makes it possible to regulate the cross-species regulation of ginseng miRNA in theory, provides ideas for ginseng miRNA to become a new pharmacodynamic substance.
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Metabolismo Energético , MicroARNs , Panax , Animales , Panax/genética , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Metabolismo Energético/genética , Células RAW 264.7 , Masculino , Fatiga/genética , Hígado/metabolismo , Hígado/inmunología , Glucógeno/metabolismoRESUMEN
Neonatal respiratory distress syndrome (NRDS) is one of the most severe respiratory disorders in preterm infants (PTIs) due to immature lung development. To delineate the serum metabolic alterations and gut microbiota variations in NRDS and assess their implications on neonatal development, we enrolled 13 NRDS neonates and 12 PTIs and collected fecal and serum specimens after birth. Longitudinal fecal sampling was conducted weekly for a month in NRDS neonates. NRDS neonates were characterized by notably reduced gestational ages and birth weights and a higher rate of asphyxia at birth relative to PTIs. Early postnatal disturbances in tryptophan metabolism were evident in the NRDS group, concomitant with elevated relative abundance of Haemophilus, Fusicatenibacter, and Vibrio. Integrative multiomics analyses revealed an inverse relationship between tryptophan concentrations and Blautia abundance. At one-week old, NRDS neonates exhibited cortisol regulation anomalies and augmented hepatic catabolism. Sequential microbial profiling revealed distinct gut microbiota evolution in NRDS subjects, characterized by a general reduction in potentially pathogenic bacteria. The acute perinatal stress of NRDS leads to mitochondrial compromise, hormonal imbalance, and delayed gut microbiota evolution. Despite the short duration of NRDS, its impact on neonatal development is significant and requires extended attention.
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Heces , Microbioma Gastrointestinal , Recien Nacido Prematuro , Síndrome de Dificultad Respiratoria del Recién Nacido , Humanos , Recién Nacido , Síndrome de Dificultad Respiratoria del Recién Nacido/microbiología , Síndrome de Dificultad Respiratoria del Recién Nacido/metabolismo , Heces/microbiología , Femenino , Masculino , Edad Gestacional , Triptófano/metabolismo , Triptófano/sangre , Hidrocortisona/sangreRESUMEN
In animal assisted reproductive technology, the production of high-quality oocytes is crucial. The yak, having lived in the Qinghai-Tibet Plateau for an extended period, has reproductive cells that are regulated by hypoxia-inducible factor 1α (HIF-1α). This study aimed to investigate the impact of HIF-1α on yak oocyte maturation and early embryonic development in vitro through the regulation of autophagy. The in vitro maturation process of yak oocytes involved the addition of the HIF-1α inducer DFOM and the inhibitor LW6 to examine their effects on yak oocyte maturation, early embryonic development, cell autophagy, cytochrome P450s (CYP450s) enzyme expression, and cumulus diffusion factors. The findings revealed that DFOM significantly upregulated the expression of HIF-1α, resulting in increased the cumulus diffusion area, elevated first polar body expulsion rate of oocytes, enhanced mitochondrial and actin levels, decreased ROS production, and reduced early apoptosis levels of oocytes. Moreover, DFOM promoted the expression of autophagy-related proteins, CYP450s enzymes, and cumulus diffusion factors, thereby enhancing oocyte maturation and early embryonic development. Conversely, LW6 exhibited opposite effects. The inhibition of autophagy levels with 3-MA during DFOM treatment yielded similar outcomes. Furthermore, reducing autophagy led to increased apoptosis levels at all stages of early embryonic development, as well as a significant decrease in total cell number and ICM/TE ratio of blastocysts. Studies have shown that during the in vitro maturation of yak oocytes, HIF-1α can affect the cumulus expansion area of oocytes by regulating autophagy, the first polar body excretion rate, mitochondrial level, actin level, ROS and early apoptosis level, the CYP450s enzyme, and the expression of cumulus expansion factors, thereby improving the in vitro maturation and early embryonic development of yak oocytes. These findings offer valuable insights into the reproductive regulation mechanism of yaks in hypoxic environments and suggest potential strategies for the advancement of yak assisted reproductive technology.
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Together, tumor and virus-specific tissue-resident CD8+ memory T cells (TRMs) of hepatocellular carcinoma (HCC) patients with Hepatitis B virus (HBV) infection can provide rapid frontline immune surveillance. The quantity and activity of CD8+ TRMs were correlated with the relapse-free survival of patients with improved health. However, HBV-specific CD8+ TRMs have a more exhausted phenotype and respond more actively under anti-PDL1 or PD1 treatment of HBV+HCC patients. Vaccination strategies that induce a strong and sustained CD8+ TRMs response are quite promising. Herein, a biodegradable poly(D,L-lactide-co-glycolide) microsphere and nanosphere particle (PLGA N.M.P) delivery system co-assembled by anti-PD1 antibodies (aPD1) and loaded with ovalbumin (OVA-aPD1 N.M.P) was fabricated and characterized for size (200 nm and 1 µm diameter), charge (-15 mV), and loading efficiencies of OVA (238 µg mg-1 particles) and aPD1 (40 µg mg-1 particles). OVA-aPD1 N.M.P could stimulate the maturation of BMDCs and enhance the antigen uptake and presentation by 2-fold compared to free OVA. The nanoparticles also induced the activation of macrophages (RAW 264.7) to produce a high level of cytokines, including TNF-α, IL-6 and IL-10. In vivo stimulation of mice using OVA-aPD1 N.M.P robustly enhanced IFN-γ-producing-CD8+ T cell infiltration in tumor tissues and the secretion of IgG and IgG2a/IgG1 antibodies. OVA-aPD1 N.M.P delivered OVA to increase the activation and proliferation of OVA-specific CD8+ TRMs, and its combination with anti-PD1 antibodies promoted complete tumor rejection by the reversal of tumor-infiltrating CD8+ T cell exhaustion. Thus, PLGA N.M.P could induce a strong CD8+ TRMs response, further highlighting its therapeutic potential in enhancing an antitumor immune response.
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Linfocitos T CD8-positivos , Carcinoma Hepatocelular , Neoplasias Hepáticas , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Animales , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/metabolismo , Copolímero de Ácido Poliláctico-Ácido Poliglicólico/química , Carcinoma Hepatocelular/inmunología , Carcinoma Hepatocelular/patología , Carcinoma Hepatocelular/terapia , Ratones , Neoplasias Hepáticas/inmunología , Neoplasias Hepáticas/patología , Neoplasias Hepáticas/terapia , Ratones Endogámicos C57BL , Ovalbúmina/inmunología , Ovalbúmina/química , Receptor de Muerte Celular Programada 1/inmunología , Receptor de Muerte Celular Programada 1/metabolismo , Nanopartículas/química , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/química , Células T de Memoria/inmunología , Vacunación , Humanos , Células RAW 264.7 , Memoria InmunológicaRESUMEN
The 2015 Paris Agreement has set out the climate change target of limiting global warming to 1.5 °C, which poses a serious challenge to countries to reduce emissions. As the world's largest carbon emitter, promoting the realization of the "dual-carbon" goal is the key to realizing China's green transformation and high-quality development. Chinese asset managers play active roles in the capital market as an important channel of asset allocation. Currently, the vast majority of Chinese asset managers hold high percentages of high-carbon industries in their portfolios, and lack quantitative data of their carbon footprints embodied in equity investments, which faces huge carbon-related risks. Therefore, it's an urgent need to comprehensively and scientifically measure financed emissions of Chinese asset managers, which is of great significance for asset managers' carbon risk management and sustainable investment. This paper develops a detailed inventory of carbon emissions for equity portfolios managed by Chinese asset managers from 2010 to 2020, which stands as a pivotal reference for in-depth analysis of emission characteristics.
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Mammalian embryos produced in vitro have poor embryo quality and low developmental ability compared with in vivo embryos. The main manifestations are the low number of blastocysts, the low ratio of the number of inner cell mass cells to the number of trophoblastic cells, and the high apoptosis rate of blastocysts, resulting in low embryo implantation rate. Therefore, optimizing in vitro culture conditions has become a key technology to im-prove the quality of preimplantation embryos. Oviduct Epithelial cells exosomes (OEVs) can be absorbed and internalized by embryos to improve the blastocyst rate and blastocyst quality of embryos in vitro. As a special nuclear structure, Paraspeckles are involved in the fate determination of mammalian early embryonic mammalian cells. However, the regulation of embryonic cell differentiation by OEVs remains unknown. We aimed to investigate the effects of OEVs on paraspeckle formation and cell fate determination in yak in vitro fertilization (IVF) of em-bryos. To simulate the in vivo oviduct environment after ovulation, we used follicular fluid exosomes (FEVs) to stimulate yak oviduct epithelial cells and collect OEVs. OEVs were added to the yak IVF embryo culture system. Paraspeckle formation, cell differentiation, and blastocyst quality in yak embryos were determined. Our results show that, development of yak embryos is unique compared to other bovine species, and OEVs can be used as a supplement to the in vitro culture system of yak embryos to improve embryonic development and blas-tocyst quality. And also Paraspeckles/CARM1 mediated the regulation of OEVs on cell differentiation during in vitro yak embryo production. These results provide new insights into the study of yak embryonic development and the role of OEVs in embryonic development.
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Diferenciación Celular , Técnicas de Cultivo de Embriones , Desarrollo Embrionario , Células Epiteliales , Exosomas , Animales , Femenino , Desarrollo Embrionario/fisiología , Bovinos/embriología , Células Epiteliales/fisiología , Células Epiteliales/metabolismo , Técnicas de Cultivo de Embriones/veterinaria , Exosomas/metabolismo , Fertilización In Vitro/veterinaria , Trompas Uterinas/citología , Blastocisto/fisiología , OviductosRESUMEN
Seed dormancy is a life adaptation trait exhibited by plants in response to environmental changes during their growth and development. The dormancy of commercial seeds is the key factor affecting seed quality. Eggplant seed dormancy is controlled by quantitative trait loci (QTLs), but reliable QTLs related to eggplant dormancy are still lacking. In this study, F2 populations obtained through the hybridization of paternally inbred lines with significant differences in dormancy were used to detect regulatory sites of dormancy in eggplant seeds. Three QTLs (dr1.1, dr2.1, and dr6.1) related to seed dormancy were detected on three chromosomes of eggplant using the QTL-Seq technique. By combining nonsynonymous sites within the candidate regions and gene functional annotation analysis, nine candidate genes were selected from three QTL candidate regions. According to the germination results on the eighth day, the male parent was not dormant, but the female parent was dormant. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the expression of nine candidate genes, and the Smechr0201082 gene showed roughly the same trend as that in the phenotypic data. We proposed Smechr0201082 as the potential key gene involved in regulating the dormancy of eggplant seeds. The results of seed experiments with different concentrations of gibberellin A3 (GA3) showed that, within a certain range, the higher the gibberellin concentration, the earlier the emergence and the higher the germination rate. However, higher concentrations of GA3 may have potential effects on eggplant seedlings. We suggest the use of GA3 at a concentration of 200-250 mg·L-1 to treat dormant seeds. This study provides a foundation for the further exploration of genes related to the regulation of seed dormancy and the elucidation of the molecular mechanism of eggplant seed dormancy and germination.
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Germinación , Latencia en las Plantas , Sitios de Carácter Cuantitativo , Semillas , Solanum melongena , Solanum melongena/genética , Solanum melongena/crecimiento & desarrollo , Sitios de Carácter Cuantitativo/genética , Latencia en las Plantas/genética , Semillas/genética , Semillas/crecimiento & desarrollo , Germinación/genética , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Mapeo Cromosómico , Fenotipo , Genes de Plantas/genéticaRESUMEN
Proteins containing domain of unknown function (DUF) are prevalent in eukaryotic genome. The DUF1216 proteins possess a conserved DUF1216 domain resembling to the mediator protein of Arabidopsis RNA polymerase II transcriptional subunit-like protein. The DUF1216 family are specifically existed in Brassicaceae, however, no comprehensive evolutionary analysis of DUF1216 genes have been performed. We performed a first comprehensive genome-wide analysis of DUF1216 proteins in Brassicaceae. Totally 284 DUF1216 genes were identified in 27 Brassicaceae species and classified into four subfamilies on the basis of phylogenetic analysis. The analysis of gene structure and conserved motifs revealed that DUF1216 genes within the same subfamily exhibited similar intron/exon patterns and motif composition. The majority members of DUF1216 genes contain a signal peptide in the N-terminal, and the ninth position of the signal peptide in most DUF1216 is cysteine. Synteny analysis revealed that segmental duplication is a major mechanism for expanding of DUF1216 genes in Brassica oleracea, Brassica juncea, Brassica napus, Lepidium meyneii, and Brassica carinata, while in Arabidopsis thaliana and Capsella rubella, tandem duplication plays a major role in the expansion of the DUF1216 gene family. The analysis of Ka/Ks (non-synonymous substitution rate/synonymous substitution rate) ratios for DUF1216 paralogous indicated that most of gene pairs underwent purifying selection. DUF1216 genes displayed a specifically high expression in reproductive tissues in most Brassicaceae species, while its expression in Brassica juncea was specifically high in root. Our studies offered new insights into the phylogenetic relationships, gene structures and expressional patterns of DUF1216 members in Brassicaceae, which provides a foundation for future functional analysis.
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Arabidopsis , Brassicaceae , Brassicaceae/genética , Duplicación de Gen , Filogenia , Evolución Molecular , Genoma de Planta , Arabidopsis/genética , Proteínas de Plantas/genética , Proteínas de Plantas/química , Planta de la Mostaza/genética , Señales de Clasificación de Proteína/genética , Regulación de la Expresión Génica de las PlantasRESUMEN
Granulosa cells (GCs) are essential for follicular development, and long non-coding RNAs (LncRNAs) are known to support the maintenance of this process and hormone synthesis in mammals. Nevertheless, the regulatory roles of these lncRNAs within sheep follicular GCs remain largely unexplored. This study delved into the influence of a Loc105611671, on the proliferation and steroid hormone synthesis of sheep ovarian GCs and the associated target genes in vitro. Cell Counting Kit-8 (CCK-8) gain-of-function experiments indicated that overexpression of Loc105611671 significantly boosted GCs proliferation, along with estrogen (E2) and progesterone (P4) levels. Further mechanistic scrutiny revealed that Loc105611671 is primarily localized within the cytoplasm of ovarian granulosa cells and engages in molecular interplay with CDC42. This interaction results in the upregulation of CDC42 protein expression. Moreover, it was discerned that increased CDC42 levels contribute to augmented proliferation of follicular granulosa cells and the secretion of E2 and P4. Experiments involving co-transfection elucidated that the concurrent overexpression of CDC42 and Loc105611671 acted synergistically to potentiate these effects. These findings provide insights into the molecular underpinnings of fecundity in ovine species and may inform future strategies for enhancing reproductive outcomes.
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AIMS: To facilitate drug discovery and development for the treatment of osteoporosis. BACKGROUND: With global aging, osteoporosis has become a common problem threatening the health of the elderly. It is of important clinical value to explore new targets for drug intervention and develop promising drugs for the treatment of osteoporosis. OBJECTIVE: To understand the major molecules that mediate the communication between the cell populations of bone marrow-derived mesenchymal stem cells (BM-MSCs) in osteoporosis and osteoarthritis patients and identify potential reusable drugs for the treatment of osteoporosis. METHODS: Single-cell RNA sequencing (scRNA-seq) data of BM-MSCs in GSE147287 dataset were classified using the Seurat package. CellChat was devoted to analyzing the ligand-receptor pairs (LR pairs) contributing to the communication between BM-MSCs subsets. The LR pairs that were differentially expressed between osteoporosis samples and control samples and significantly correlated with immune score were screened in the GSE35959 dataset, and the differentially expressed gene in both GSE35959 and GSE13850 data sets were identified as targets from a single ligand or receptor. The therapeutic drugs for osteoporosis were screened by network proximity method, and the top-ranked drugs were selected for molecular docking and molecular dynamics simulation with the target targets. RESULTS: Twelve subsets of BM-MSCs were identified, of which CD45-BM-MSCS_4, CD45-BM- MSCS_5, and CD45+ BM-MSCs_5 subsets showed significantly different distributions between osteoporosis samples and osteoarthritis samples. Six LR pairs were identified in the bidirectional communication between these three BM-MSCs subsets and other BM-MSCs subsets. Among them, MIF-CD74 and ITGB2-ICAM2 were significantly correlated with the immune score. CD74 was identified as the target, and a total of 48 drugs targeting CD47 protein were identified. Among them, DB01940 had the lowest free energy binding score with CD74 protein and the binding state was very stable. CONCLUSION: This study provided a new network-based framework for drug reuse and identified initial insights into therapeutic agents targeting CD74 in osteoporosis, which may be meaningful for promoting the development of osteoporosis treatment.
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An appropriate water supply strategy is imperative for obtaining tomatoes of a high yield and quality; the lack of one has caused resource wastage and quality deterioration. To determine the suitable irrigation amount and simulate daily transpiration under these optimal irrigation conditions, a two-year greenhouse cultivation experiment was conducted over 2022-2023. Commencing at anthesis, three distinct irrigation gradients were triggered and designated as irrigation controls with the lower limits set at 80% (T1), 70% (T2), and 60% (T3) of the substrate water-holding capacity. We determined the optimal irrigation amount by ranking the treatments using the TOPSIS method, balancing the tomato yield and quality. A segmented daily transpiration model under optimal irrigation conditions driven by crop and environmental factors was established using the Marquardt method and data from 2022, and the model was validated using data from 2023. The results indicated that T2 was the optimal irrigation amount, with the water use efficiency increased by 18.0%, but with a 10.9% decrease in yield, while the quality indices improved significantly. The R2 values of the segmented model in the flowering and fruit-setting stage and the picking stage were 0.92 and 0.86, respectively, which could provide support for optimized water management for tomato planting in greenhouse substrate cultivation.
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Radish (Raphanus sativus L.) is a vegetable crop with economic value and ecological significance in the genus Radish, family Brassicaceae. In recent years, developed countries have attached great importance to the collection and conservation of radish germplasm resources and their research and utilization, but the lack of population genetic information and molecular markers has hindered the development of the genetic breeding of radish. In this study, we integrated the radish genomic data published in databases for the development of single-nucleotide polymorphism (SNP) markers, and obtained a dataset of 308 high-quality SNPs under strict selection criteria. With the support of Kompetitive Allele-Specific PCR (KASP) technology, we screened a set of 32 candidate core SNP marker sets to analyse the genetic diversity of the collected 356 radish varieties. The results showed that the mean values of polymorphism information content (PIC), minor allele frequency (MAF), gene diversity and heterozygosity of the 32 candidate core SNP markers were 0.32, 0.30, 0.40 and 0.25, respectively. Population structural analysis, principal component analysis and genetic evolutionary tree analysis indicated that the 356 radish materials were best classified into two taxa, and that the two taxa of the material were closely genetically exchanged. Finally, on the basis of 32 candidate core SNP markers we calculated 15 core markers using a computer algorithm to construct a fingerprint map of 356 radish varieties. Furthermore, we constructed a core germplasm population consisting of 71 radish materials using 32 candidate core markers. In this study, we developed SNP markers for radish cultivar identification and genetic diversity analysis, and constructed DNA fingerprints, providing a basis for the identification of radish germplasm resources and molecular marker-assisted breeding as well as genetic research.
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As one of the endocrine-disrupting chemicals (EDCs), dibutyl phthalate (DBP) has been extensively used in industry. DBP has been shown to cause damage to Leydig cells, yet its underlying mechanism remains elusive. In this study, we show that DBP induces ferroptosis of mouse Leydig cells via upregulating the expression of Sp2, a transcription factor. Also, Sp2 is identified to promote the transcription of Vdac2 gene by binding to its promoter and subsequently involved in DBP-induced ferroptosis of Leydig cells. In addition, DBP is proved to induce ferroptosis via inducing oxidative stress, while inhibition of oxidative stress by melatonin alleviates DBP-induced ferroptosis and upregulation of Sp2 and VDAC2. Taken together, our findings demonstrate that melatonin can alleviate DBP-induced ferroptosis of mouse Leydig cells via inhibiting oxidative stress-triggered Sp2/VDAC2 signals.
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Ferroptosis , Melatonina , Ratones , Masculino , Animales , Dibutil Ftalato/toxicidad , Células Intersticiales del Testículo/metabolismo , Testículo/metabolismo , Melatonina/farmacología , Melatonina/metabolismoRESUMEN
INTRODUCTION: This study aimed to investigate the microbial characteristics of yak uteri collected using intrauterine cotton swabs (CS) during different reproductive stages and the correlation of these microbial characteristics with reproductive status. METHODS: We used a macrogenomic approach to analyze the functional aspects of different microorganisms in samples collected during the pre-estrus, estrus, late estrus, and diestrus stages. RESULTS: The results revealed the presence of 1293 microbial genera and 3401 microbial species in the uteri of yaks at different reproductive stages. The dominant bacterial species varied across the different periods, with Micrococcus and Proteus being dominant during pre-estrus; Pseudomonas, Clostridium, Flavobacterium, Bacillus, and Staphylococcus during estrus; Acinetobacter, Bacillus and Proteus during late estrus; and Pseudomonas, Escherichia coli, and Proteus during diestrus. DISCUSSION: The primary functions of these bacteria are enriched in various metabolic pathways, including carbohydrate and amino acid metabolism, intracellular transport and secretion, post-translational protein modification, and drug resistance. These findings suggest that the microbial diversity in the uterus of yaks plays a crucial role in reproductive regulation and can help prevent reproductive tract-related diseases.
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Estro , Útero , Femenino , Bovinos , Animales , Útero/metabolismo , ReproducciónRESUMEN
As an extensively employed plasticizer in industrial applications, di-2-ethylhexyl phthalate (DEHP) can induce apoptosis of mouse Leydig cells, yet the precise mechanism remains elusive. In the current study, we identified that DEHP could specially induced apoptosis in the Leydig cells of the testis tissue, accompanied with the upregulation of apoptosis-related protein in the TGF-ß signaling pathway (ARTS) in the cells. Overexpression of ARTS significantly induced apoptosis of TM3 cells, while knockdown of ARTS inhibited apoptosis. Furthermore, DEHP-induced apoptosis of TM3 cells could be alleviated by knockdown of ARTS, which indicated that ARTS was involved in DEHP-induced apoptosis of mouse Leydig cells. Bioinformation assay predicts that there are four potential p53-responsive elements (p53-REs) located at - 6060, - 5726, - 5631 and - 5554 before the transcription start site of ARTS gene, implying that gene transcription of ARTS could be regulated by p53. Interestingly, DEHP was shown to specifically upregulate the expression of p53 in the Leydig cells of the testis tissue and TM3 cells. Consistently, p53 was proved to bind to the RE4 site of the ARTS gene promoter and transcriptionally activated the promoter-driven expression of the luciferase reporter gene. Overexpression of p53 could induce apoptosis of TM3 cells; while knockdown of p53 could not only rescue DEHP-induced apoptosis of the cells, but also inhibit DEHP-caused upregulation of ARTS. Meanwhile, we showed that oxidative stress could induce apoptosis of TM3 cells, accompanied with the increased protein levels of p53 and ARTS; while inhibition of oxidative stress dramatically alleviated DEHP-induced apoptosis and the up-regulation of p53 and ARTS. Taken together, these results indicated that DEHP-induced oxidative stress activates the p53-ARTS cascade to promote apoptosis of mouse Leydig cells.
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Dietilhexil Ftalato , Células Intersticiales del Testículo , Ácidos Ftálicos , Ratones , Animales , Masculino , Células Intersticiales del Testículo/metabolismo , Dietilhexil Ftalato/toxicidad , Dietilhexil Ftalato/metabolismo , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismo , Apoptosis , Testículo/metabolismoRESUMEN
The deubiquitylase OTU domain-containing ubiquitin aldehyde-binding protein 1 (OTUB1) has been implicated in the pathogenesis of various human diseases. However, the molecular mechanism by which OTUB1 participates in the pathogenesis of intracerebral hemorrhage (ICH) remains elusive. In the present study, we established an autologous whole blood fusion-induced ICH model in C57BL/6 J mice. We showed that the upregulation of OTUB1 contributes to the attenuation of Nuclear factor kappa B (NF-κB) and its downstream apoptotic signaling after ICH. OTUB1 directly associates with NF-κB precursors p105 and p100 after ICH, leading to attenuated polyubiquitylation of p105 and p100. Moreover, we revealed that NF-κB signaling was modestly activated both in ICH tissues and hemin-exposed HT-22 neuronal cells, accompanied with the activation of NF-κB downstream pro-apoptotic signaling. Notably, overexpression of OTUB1 strongly inhibited hemin-induced NF-κB activation, whereas interference of OTUB1 led to the opposite effect. Finally, we revealed that lentiviral transduction of OTUB1 markedly ameliorated hemin-induced apoptotic signaling and HT-22 neuronal death. Collectively, these findings suggest that the upregulation of OTUB1 serves as a neuroprotective mechanism in antagonizing neuroinflammation-induced NF-κB signaling and neuronal death, shed new light on manipulating intracellular deubiquitylating pathways as novel interventive approaches against ICH-induced secondary neuronal damage and death.
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Hemina , FN-kappa B , Animales , Humanos , Ratones , Hemorragia Cerebral/patología , Hemina/farmacología , Ratones Endogámicos C57BL , FN-kappa B/metabolismo , Transducción de SeñalRESUMEN
As a regulatory protein that upregulates transcription in response to various stresses, cold-induced RNA-binding protein (CIRBP) is involved in a variety of physiological pathological processes in cells. However, little is known about the role of CIRBP in regulating autophagy and the synthesis and secretion of ovarian steroid hormones (estradiol E2 and progesterone P4). This study aimed to explore whether the synthetic secretion of ovarian steroid hormones is related to CIRBP-regulated autophagy. We detected the differential expression of CIRBP, LC3, E2 and P4 in YGCs cultured at mild low temperature (32 °C) for 6 and 12 h. CIRBP, LC3, E2 and P4 expression was increased in response to low temperature in YGCs. In order to illustrate that the changes in secretion of E2/P4 and autophagy might be caused by CIRBP induced by low temperature, we overexpressed CIRBP in YGCs cultured in vitro to detect its effects on autophagy and steroid hormone synthesis and secretion. We found that overexpression of CIRBP can induce autophagy of YGCs and enhance the synthesis and secretion of E2 and P4, suggesting that mild hypothermia may activate autophagy by inducing the expression of CIRBP and enhance the synthesis and secretion of E2 and P4. To further explore the relationship between CIRBP regulated autophagy and steroid hormone synthesis and secretion, we verified it by regulating autophagy. The results showed that Inhibition of autophagy significantly reversed CIRBP overexpression-enhanced autophagy and synthetic secretion of E2, P4 in YGCs, while activated autophagy showed similar results to overexpression of CIRBP. In conclusion, our data suggest that autophagy is involved in the synthesis and secretion of YGCs E2 and P4 and is associated with overexpression of CIRBP.
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Células de la Granulosa , Progesterona , Animales , Bovinos , Femenino , Progesterona/metabolismo , Células de la Granulosa/metabolismo , Estradiol/metabolismoRESUMEN
As one of the most important phthalates, di-isononyl phthalate (DINP) has been widely used as a common plasticizer in the food and personal care products sectors. In our previous study, we found that DINP can induce autophagy of ovarian granulosa cells; while the underlying mechanism is unclear. In the study, we showed that DINP exposure could induce autophagy of ovarian granulosa cells and KGN cells, accompanied with the increase in the mRNA and protein level of DDIT4. Furthermore, overexpression of DDIT4 were shown to induce autophagy of KGN cells; while knockdown of DDIT4 inhibited DINP-induced autophagy, implying that DDIT4 played an important role in DINP-induced autophagy of ovarian granulosa cells. There were three putative binding sites of transcription factor ATF4 in the promoter region of DDIT4 gene, suggesting that DDIT4 might be regulated by ATF4. Herein, we found that overexpression of ATF4 could upregulate the expression of DDIT4 in KGN cells, while knockdown of ATF4 inhibited its expression. Subsequently, ATF4 was identified to bind to the promoter region of DDIT4 gene and promote its transcription. The expression of ATF4 was also increased in the DINP-exposed granulosa cells, and ATF4 overexpression promoted autophagy of KGN cells; whereas knockdown of ATF4 alleviated DINP-induced upregulation of DDIT4 and autophagy of the cells. Taken together, DINP triggered autophagy of ovarian granulosa cells through activating ATF4/DDIT4 signals.
Asunto(s)
Regulación de la Expresión Génica , Ácidos Ftálicos , Femenino , Humanos , Ácidos Ftálicos/química , Autofagia/genética , Células de la GranulosaRESUMEN
Understanding the microflora inhabiting the reproductive tract is important for a better understanding of female physiology and reproductive health. The endometrial fluid from mice in three reproductive stages (A: Unproductive mice; B: Postovulatory mice; C: Postpartum mice) was extracted for microbial DNA extraction and sequencing. Phenotypic and functional analyses of endometrial microbial enrichment was undertaken using LefSe. The results showed 95 genera and 134 species of microorganisms in the uteri of mice. There were differentially distributed genera, among which Lactobacillus, Enterococcus, and Streptococcus were more abundant in the endometrial fluid of mice in the unproductive group. That of mice in the postovulatory group was colonized with Salmonella enterica and Campylobacter and was mainly enriched in metabolic pathways and steroid biosynthesis. The presence of Chlamydia, Enterococcus, Pseudomonadales, Acinetobacter, and Clostridium in the endometrial fluid of postpartum mice, in addition to the enrichment of the endocrine system and the Apelin and FoxO signaling pathways, resulted in a higher number of pathogenic pathways than in the other two groups. The results showed that the microbial diversity characteristics in the endometrium of mice in different reproductive states differed and that they could be involved in the regulation of animal reproduction through metabolic pathways and steroid biosynthesis, suggesting that reproductive diseases induced by microbial diversity alterations in the regulation of animal reproduction cannot be ignored.