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1.
Nat Commun ; 12(1): 1758, 2021 03 19.
Artículo en Inglés | MEDLINE | ID: mdl-33741948

RESUMEN

The molecular machinery and chromosome structures carrying out meiosis are frequently conserved from yeast to mammals. However, signals initiating meiosis appear divergent: while nutrient restriction induces meiosis in the yeast system, retinoic acid (RA) and its target Stra8 have been shown to be necessary but not sufficient to induce meiotic initiation in mammalian germ cells. Here, we use primary culture of mouse undifferentiated spermatogonia without the support of gonadal somatic cells to show that nutrient restriction in combination with RA is sufficient to induce Stra8- and Spo11-dependent meiotic gene and chromosome programs that recapitulate the transcriptomic and cytologic features of in vivo meiosis. We demonstrate that neither nutrient restriction nor RA alone exerts these effects. Moreover, we identify a distinctive network of 11 nutrient restriction-upregulated transcription factor genes, which are associated with early meiosis in vivo and whose expression does not require RA. Our study proposes a conserved model, in which nutrient restriction induces meiotic initiation by upregulating key transcription factor genes for the meiotic gene program and provides an in vitro platform for meiotic induction that could facilitate research and haploid gamete production.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Endodesoxirribonucleasas/metabolismo , Meiosis/efectos de los fármacos , Nutrientes/metabolismo , Espermatogonias/efectos de los fármacos , Tretinoina/farmacología , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Células Cultivadas , Endodesoxirribonucleasas/genética , Expresión Génica/efectos de los fármacos , Masculino , Mamíferos/genética , Mamíferos/metabolismo , Meiosis/genética , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Noqueados , Análisis de Secuencia de ARN/métodos , Análisis de la Célula Individual/métodos , Espermatogonias/citología , Espermatogonias/metabolismo
2.
Environ Health Prev Med ; 26(1): 34, 2021 Mar 11.
Artículo en Inglés | MEDLINE | ID: mdl-33706700

RESUMEN

BACKGROUND: Arsenic is a developmental neurotoxicant. It means that its neurotoxic effect could occur in offspring by maternal arsenic exposure. Our previous study showed that developmental arsenic exposure impaired social behavior and serotonergic system in C3H adult male mice. These effects might affect the next generation with no direct exposure to arsenic. This study aimed to detect the social behavior and related gene expression changes in F2 male mice born to gestationally arsenite-exposed F1 mice. METHODS: Pregnant C3H/HeN mice (F0) were given free access to tap water (control mice) or tap water containing 85 ppm sodium arsenite from days 8 to 18 of gestation. Arsenite was not given to F1 or F2 mice. The F2 mice were generated by mating among control F1 males and females, and arsenite-F1 males and females at the age of 10 weeks. At 41 weeks and 74 weeks of age respectively, F2 males were used for the assessment of social behavior by a three-chamber social behavior apparatus. Histological features of the prefrontal cortex were studied by ordinary light microscope. Social behavior-related gene expressions were determined in the prefrontal cortex by real time RT-PCR method. RESULTS: The arsenite-F2 male mice showed significantly poor sociability and social novelty preference in both 41-week-old group and 74-week-old group. There was no significant histological difference between the control mice and the arsenite-F2 mice. Regarding gene expression, serotonin receptor 5B (5-HT 5B) mRNA expression was significantly decreased (p < 0.05) in the arsenite-F2 male mice compared to the control F2 male mice in both groups. Brain-derived neurotrophic factor (BDNF) and dopamine receptor D1a (Drd1a) gene expressions were significantly decreased (p < 0.05) only in the arsenite-F2 male mice of the 74-week-old group. Heme oxygenase-1 (HO-1) gene expression was significantly increased (p < 0.001) in the arsenite-F2 male mice of both groups, but plasma 8-hydroxy-2'-deoxyguanosine (8-OHdG) and cyclooxygenase-2 (COX-2) gene expression were not significantly different. Interleukin-1ß (IL-1ß) mRNA expression was significantly increased only in 41-week-old arsenite-F2 mice. CONCLUSIONS: These findings suggest that maternal arsenic exposure affects social behavior in F2 male mice via serotonergic system in the prefrontal cortex. In this study, COX-2 were not increased although oxidative stress marker (HO-1) was increased significantly in arsnite-F2 male mice.


Asunto(s)
Arsénico/toxicidad , Arsenitos/toxicidad , Contaminantes Ambientales/toxicidad , Expresión Génica/efectos de los fármacos , Exposición Materna/efectos adversos , Efectos Tardíos de la Exposición Prenatal/inducido químicamente , Conducta Social , Compuestos de Sodio/toxicidad , Animales , Conducta Animal/efectos de los fármacos , Femenino , Marcadores Genéticos , Masculino , Ratones , Ratones Endogámicos C3H , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/genética , Corteza Prefrontal/efectos de los fármacos , Embarazo , Efectos Tardíos de la Exposición Prenatal/genética , Efectos Tardíos de la Exposición Prenatal/metabolismo , Efectos Tardíos de la Exposición Prenatal/psicología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Serotonina/genética , Serotonina/metabolismo
3.
Arch Insect Biochem Physiol ; 106(4): e21772, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33719088

RESUMEN

The ß-adrenergic-like octopamine receptor (OA2B2), which binds the biogenic amine octopamine, belongs to the class of G-protein coupled receptors and significantly regulates many physiological and behavioral processes in insects. In this study, the putative open reading frame sequence of the MsOA2B2 gene in Mythimna separata was cloned, the full-length complementary DNA was 1191 bp and it encoded a 396-amino acid protein (GenBank accession number MN822800). Orthologous sequence alignment, phylogenetic tree analysis, and protein sequence analysis all showed that the cloned receptor belongs to the OA2B2 protein family. Real-time quantitative polymerase chain reaction of spatial and temporal expression analysis revealed that the MsOAB2 gene was expressed in all developmental stages of M. separata and was most abundant in egg stages and second and fourth instars compared with other developmental stages, while the expression level during the pupal stage was much lower than that at the other stages. Further analysis with sixth instar M. separata larvae showed that the MsOA2B2 gene was expressed 1.81 times higher in the head than in integument and gut tissues. Dietary ingestion of dsMsOA2B2 significantly reduced the messenger RNA level of MsOA2B2 and decreased mortality following amitraz treatment. This study provides both a pharmacological characterization and the gene expression patterns of OA2B2 in M. separata, facilitating further research for insecticides using MsOA2B2 as a target.


Asunto(s)
Mariposas Nocturnas/genética , Receptores de Amina Biogénica , Animales , Expresión Génica/efectos de los fármacos , Genes de Insecto , Control de Insectos , Proteínas de Insectos/química , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Insecticidas/farmacología , Larva/genética , Larva/metabolismo , Mariposas Nocturnas/metabolismo , Filogenia , Pupa/genética , Pupa/metabolismo , Receptores Adrenérgicos beta/química , Receptores Adrenérgicos beta/efectos de los fármacos , Receptores Adrenérgicos beta/genética , Receptores Adrenérgicos beta/metabolismo , Receptores de Amina Biogénica/química , Receptores de Amina Biogénica/efectos de los fármacos , Receptores de Amina Biogénica/genética , Receptores de Amina Biogénica/metabolismo , Toluidinas/farmacología
4.
Aquat Toxicol ; 233: 105788, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33662878

RESUMEN

The gene expression response thought to underlie the negative apical effects resulting from estrogen exposure have been thoroughly described in fish. Although epigenetics are believed to play a critical role translating environmental exposures into the development of adverse apical effects, they remain poorly characterized in fish species. This study investigated alterations of DNA methylation of estrogen receptor alpha (esr1) in brain and liver tissues from 8 to 10 month old male fathead minnows (Pimephales promelas) after a 2d exposure to either 2.5 ng/L or 10 ng/L 17α-ethynylestradiol (EE2). Changes in the patterns of methylation were evaluated using targeted deep sequencing of bisulfite treated DNA in the 5' region of esr1. Methylation and gene expression were assessed at 2d of exposure and after a 7 and 14d depuration period. After 2d EE2 exposure, males exhibited significant demethylation in the 5' upstream region of esr1 in liver tissue, which was inversely correlated to gene expression. This methylation pattern reflected what was seen in females. No gene body methylation (GBM) was observed for liver of exposed males. Differential methylation was observed for a single upstream CpG site in the liver after the 14d depuration. A less pronounced methylation response was observed in the upstream region in brain tissue, however, several CpGs were necessarily excluded from the analysis. In contrast to the liver, a significant GBM response was observed across the entire gene body, which was sustained until at least 7d post-exposure. No differential expression was observed in the brain, limiting functional interpretation of methylation changes. The identification of EE2-dependent changes in methylation levels strongly suggests the importance of epigenetic mechanisms as a mediator of the organismal response to environmental exposures and the need for further characterization of the epigenome. Further, differential methylation following depuration indicates estrogenic effects persist well after the active exposure, which has implications for the risk posed by repeated exposures..


Asunto(s)
Cyprinidae/metabolismo , Metilación de ADN/efectos de los fármacos , Receptor alfa de Estrógeno/genética , Etinilestradiol/toxicidad , Expresión Génica/efectos de los fármacos , Contaminantes Químicos del Agua/toxicidad , Animales , Encéfalo/efectos de los fármacos , Encéfalo/metabolismo , Cyprinidae/genética , Estrógenos/metabolismo , Femenino , Hígado/efectos de los fármacos , Hígado/metabolismo , Masculino , Vitelogeninas/metabolismo
5.
Aquat Toxicol ; 233: 105783, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33662881

RESUMEN

Pesticides occur in the environment as mixtures, yet the joint toxicity of pesticide mixtures remains largely under-explored and is usually overlooked in ecological risk assessment. In the current study, joint toxicity of a neonicotinoid insecticide (imidacloprid, IMI) and a strobilurin fungicide (azoxystrobin, AZO) was investigated with Chironomus dilutus over a wide range of concentrations and at different effect levels (organism, cell, and gene levels). The two pesticides, both individually and in combination, were found to induce oxidative stress and cause lethality in C. dilutus. Median lethal concentrations for IMI and AZO were 3.98 ± 1.17 and 52.9 ± 1.1 µg/L, respectively. Mixtures of the two pesticides presented synergetic effects at environmentally relevant concentrations whilst antagonistic effects at high concentrations, showing concentration-dependent joint toxicity. Investigation on the expressions of 12 genes (cyt b, coi, cox1, cyp4, cyp12m1, cyp9au1, cyp6fv1, cyp315, gst, Zn/Cu-sod, Mn-sod, and cat) revealed that the two pesticides impaired mitochondrial respiration, detoxification, and antioxidant system of C. dilutus, and the joint effects of the two pesticides were likely due to an interplay between their respective influences on these physiological processes. Collectively, the synergistic effects of the two pesticides at environmentally relevant concentrations highlight the importance to incorporate combined toxicity studies into ecological risk assessment of pesticides.


Asunto(s)
Chironomidae/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Neonicotinoides/toxicidad , Nitrocompuestos/toxicidad , Plaguicidas/toxicidad , Pirimidinas/toxicidad , Estrobilurinas/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Chironomidae/citología , Chironomidae/genética , Sinergismo Farmacológico , Peróxido de Hidrógeno/metabolismo , Dosificación Letal Mediana , Malondialdehído/metabolismo , Modelos Teóricos
6.
Aquat Toxicol ; 233: 105789, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33667915

RESUMEN

Fluoride has been found to cause detrimental effects on fish gills. Despite essential roles in various metabolism activities, whether and how miRNAs participate in the course of toxicity caused by fluoride in gills is still unclear. In this study, male zebrafish were exposed to 0, 20, 40 mg/L fluoride for 60 days to study the underlying osmotic regulatory mechanism by determining the influences of fluoride on the miRNAs and regulated genes in gills. mRNAs were isolated from the gills and the expression profiles were analyzed by using Illumina Hiseq 2500 platforms. Expressions of 7 differentially miRNAs and some related-genes in gills were validated by qRT-PCR. The results showed that miRNAs expressions were notably altered by fluoride. A total of 584 and 327 miRNAs were remarkably changed after 20 and 40 mg/L fluoride exposure, of which 322 were increased and 262 were decreased in 20 mg/L fluoride group, whereas 219 were elevated and 108 were reduced in 40 mg/L fluoride group. The differentially expressive miRNAs confirmed by qRT-PCR were consistent with micro-assay data. Cluster of Orthologous Groups of proteins (COG) function classification showed that the target genes of differentially expressive miRNAs are mainly related to signal transduction mechanisms, replication, transcription, inorganic ion transport and metabolism, repair and recombination, and energy formation and transformation. In addition, fluoride disturbed the expressions of target genes involved in the osmoregulation of the gill in the fluoride-exposed zebrafish, such as the increased expressions of OSTF1 and the decreased expressions of Na+-K+-ATPase, CFTR, and AQP-3, which provides a possibility that miRNAs regulation induced by fluoride has an effects on osmotic regulation, providing new hints to the osmotic regulatory mechanism of the toxicity caused by fluoride in zebrafish, and distinguishes new biomarkers of miRNAs for fluoride toxicity.


Asunto(s)
Fluoruros/toxicidad , Expresión Génica/efectos de los fármacos , Branquias/efectos de los fármacos , MicroARNs/metabolismo , Contaminantes Químicos del Agua/toxicidad , Pez Cebra/metabolismo , Animales , Branquias/metabolismo , Secuenciación de Nucleótidos de Alto Rendimiento , Masculino , MicroARNs/genética , Pez Cebra/genética
7.
Gene ; 781: 145488, 2021 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-33588040

RESUMEN

Oxidative stress (OS) plays an essential role in demyelination and tissue injury related to pathogenesis of multiple sclerosis (MS). On the other hand, vitamin D (VD) as an antioxidant reduces oxidative stress and has been used as adjuvant therapy in autoimmune diseases. Although VD supplementation is suggested as a protective and immunomodulation factor for MS patients, the molecular mechanisms remain unclear. Given that VD may modulate the immune system of MS patients through the DNA repair pathway, we aimed to evaluate the effects of VD supplementation in DNA repair genes expression including OGG1, MYH, MTH1, and ITPA. Transcript levels were measured using the RT-qPCR method in peripheral blood mononuclear cells (PBMCs) of relapsing-remitting multiple sclerosis (RRMS) patients before and after two months of VD supplementation. Furthermore, in silico analysis and correlation gene expression analysis was performed to find the biological binding sites and the effect of NRF2 on the regulation of DNA repair genes. Our data revealed that in MS patients, 2-month VD treatment significantly altered the expression of MYH, OGG1, MTH1, and NRF2 genes. A significant correlation was observed between DNA repair genes and NRF2 expression, which was confirmed by the presence of antioxidant response element (ARE) binding sites in the promoter of OGG1, MYH, and MTH1 genes. This study demonstrated that the impact of VD on MS patients may be mediated through the improvement of DNA repair system efficiency. This finding brought some new evidence for the involvement of DNA repair genes in the physiopathology of MS patients.


Asunto(s)
Reparación del ADN/genética , Expresión Génica/efectos de los fármacos , Esclerosis Múltiple/genética , Vitamina D/farmacología , Vitaminas/farmacología , Adulto , Simulación por Computador , ADN Glicosilasas/genética , Reparación del ADN/efectos de los fármacos , Enzimas Reparadoras del ADN/genética , Femenino , Humanos , Masculino , Esclerosis Múltiple/tratamiento farmacológico , Factor 2 Relacionado con NF-E2/genética , Monoéster Fosfórico Hidrolasas/genética , Reacción en Cadena en Tiempo Real de la Polimerasa
8.
Nat Commun ; 12(1): 773, 2021 02 03.
Artículo en Inglés | MEDLINE | ID: mdl-33536439

RESUMEN

Macrophages are plastic and, in response to different local stimuli, can polarize toward multi-dimensional spectrum of phenotypes, including the pro-inflammatory M1-like and the anti-inflammatory M2-like states. Using a high-throughput phenotypic screen in a library of ~4000 FDA-approved drugs, bioactive compounds and natural products, we find ~300 compounds that potently activate primary human macrophages toward either M1-like or M2-like state, of which ~30 are capable of reprogramming M1-like macrophages toward M2-like state and another ~20 for the reverse repolarization. Transcriptional analyses of macrophages treated with 34 non-redundant compounds identify both shared and unique targets and pathways through which the tested compounds modulate macrophage activation. One M1-activating compound, thiostrepton, is able to reprogram tumor-associated macrophages toward M1-like state in mice, and exhibit potent anti-tumor activity. Our compound-screening results thus help to provide a valuable resource not only for studying the macrophage biology but also for developing therapeutics through modulating macrophage activation.


Asunto(s)
Antiinflamatorios/farmacología , Productos Biológicos/farmacología , Ensayos Analíticos de Alto Rendimiento/métodos , Activación de Macrófagos/efectos de los fármacos , Macrófagos/efectos de los fármacos , Animales , Antiinflamatorios/química , Productos Biológicos/química , Línea Celular Tumoral , Células Cultivadas , Expresión Génica/efectos de los fármacos , Ontología de Genes , Humanos , Macrófagos/clasificación , Macrófagos/metabolismo , Ratones Endogámicos C57BL , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/patología , Neoplasias Experimentales/prevención & control , Fenotipo , Células THP-1 , Tioestreptona/química , Tioestreptona/farmacología
9.
Nat Commun ; 12(1): 866, 2021 02 08.
Artículo en Inglés | MEDLINE | ID: mdl-33558541

RESUMEN

The coronavirus disease 2019 (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has rapidly become a global public health threat. The efficacy of several repurposed drugs has been evaluated in clinical trials. Among these drugs, a second-generation antiandrogen agent, enzalutamide, was proposed because it reduces the expression of transmembrane serine protease 2 (TMPRSS2), a key component mediating SARS-CoV-2-driven entry, in prostate cancer cells. However, definitive evidence for the therapeutic efficacy of enzalutamide in COVID-19 is lacking. Here, we evaluated the antiviral efficacy of enzalutamide in prostate cancer cells, lung cancer cells, human lung organoids and Ad-ACE2-transduced mice. Tmprss2 knockout significantly inhibited SARS-CoV-2 infection in vivo. Enzalutamide effectively inhibited SARS-CoV-2 infection in human prostate cells, however, such antiviral efficacy was lacking in human lung cells and organoids. Accordingly, enzalutamide showed no antiviral activity due to the AR-independent TMPRSS2 expression in mouse and human lung epithelial cells. Moreover, we observed distinct AR binding patterns between prostate cells and lung cells and a lack of direct binding of AR to TMPRSS2 regulatory locus in human lung cells. Thus, our findings do not support the postulated protective role of enzalutamide in treating COVID-19 through reducing TMPRSS2 expression in lung cells.


Asunto(s)
/prevención & control , Especificidad de Órganos/genética , Feniltiohidantoína/análogos & derivados , Serina Endopeptidasas/genética , /genética , Animales , /virología , Línea Celular Tumoral , Células Cultivadas , Expresión Génica/efectos de los fármacos , Interacciones Huésped-Patógeno/efectos de los fármacos , Humanos , Masculino , Ratones Noqueados , Pandemias , Feniltiohidantoína/farmacología , Neoplasias de la Próstata/metabolismo , Neoplasias de la Próstata/patología , Neoplasias de la Próstata/virología , Unión Proteica/efectos de los fármacos , Serina Endopeptidasas/metabolismo
10.
Ecotoxicol Environ Saf ; 208: 111606, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-33396126

RESUMEN

Mancozeb is a metal-containing ethylene bis-dithiocarbamate fungicide widely used in agriculture. Ethylene thiourea (ETU) is the primary metabolite of Mancozeb. Mancozeb has been associated with spontaneous abortions and abnormal menstruation in women. However, the effects of Mancozeb and ETU on embryo attachment remain unknown. The human blastocyst surrogate trophoblastic spheroids (JEG-3), endometrial epithelial surrogate adenocarcinoma cells (Ishikawa), or human primary endometrial epithelial cells (EECs) monolayer were used in the spheroid attachment models. Ishikawa and EECs were pretreated with different concentrations of Mancozeb or ETU for 48 h before the attachment assay. Gene expression profiles of Ishikawa cells were examined to understand how Mancozeb modulates endometrial receptivity with Microarray. The genes altered by Mancozeb were confirmed by qPCR and compared with the ETU treated groups. Mancozeb and ETU treatment inhibited cell viability at 10 µg/mL and 5000 µg/mL, respectively. At non-cytotoxic concentrations, Mancozeb at 3 µg/mL and ETU at 300 µg/mL reduced JEG-3 spheroid attachment onto Ishikawa cells. A similar result was observed with human primary endometrial epithelial cells. Mancozeb at 3 µg/mL modified the transcription of 158 genes by at least 1.5-fold in Microarray analysis. The expression of 10 differentially expressed genes were confirmed by qPCR. Furthermore, Mancozeb decreased spheroid attachment possibly through downregulating the expression of endometrial estrogen receptor ß and integrin ß3, but not mucin 1. These results were confirmed in both overexpression and knockdown experiments and co-culture assay. Mancozeb but not its metabolite ETU reduced spheroid attachment through modulating gene expression profile and decreasing estrogen receptor ß and integrin ß3 expression of endometrial epithelial cells.


Asunto(s)
Adhesión Celular/efectos de los fármacos , Endometrio/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Receptor beta de Estrógeno/metabolismo , Fungicidas Industriales/toxicidad , Integrina beta3/metabolismo , Maneb/toxicidad , Esferoides Celulares/efectos de los fármacos , Zineb/toxicidad , Blastocisto/citología , Blastocisto/efectos de los fármacos , Blastocisto/metabolismo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Técnicas de Cocultivo , Regulación hacia Abajo , Endometrio/citología , Endometrio/metabolismo , Células Epiteliales/metabolismo , Receptor beta de Estrógeno/genética , Femenino , Expresión Génica/efectos de los fármacos , Técnicas de Silenciamiento del Gen , Humanos , Integrina beta3/genética , Embarazo , Esferoides Celulares/metabolismo
11.
Ecotoxicol Environ Saf ; 208: 111614, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-33396134

RESUMEN

A novel gill cell line from pearl gentian grouper (Epinephelus lanceolatus♂×Epinephelus fuscoguttatus♀, PGGG cell line) was established, its application in cadmium (Cd) toxicology was demonstrated in this study. Primary cultures and PGGG subcultures were carried out at 25 °C in Dulbecco's Modified Eagle medium/F12 medium (1:1; pH 7.2) supplemented with 15% fetal bovine serum (FBS). Primary PGGG cells were spindle-shaped, proliferated into a confluent monolayer within two weeks and were continuously subcultured over passage 60. The growth of cells at passages 20, 40, and 60 was examined. Chromosome analysis revealed that the chromosomal number of normal PGGG cells was 48, but the number of cells with the normal chromosomes number decreased during the passaging process. Cadmium is one of the most toxic metals in aquatic systems and has been associated with multiple animal and human health problems. To interpret the cytotoxicity and related mechanisms of cadmium, PGGG cells were used as an in vitro model. After treatment with cadmium at concentrations ranging from 1 µM to 500 µM, PGGG cells demonstrated dose- and time-dependent cytotoxicity, manifested as morphological abnormalities and a viability decline. Further, it was found that the reactive oxygen species (ROS) and malondialdehyde (MDA) levels were elevated following cadmium exposure, and related genes involved in the antioxidant system, including those encoding catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), and Kelch-like- ECH-associated protein 1 (Keap1), were regulated differently. In addition, PGGG cells treated with cadmium had the typical features associated with apoptosis, including phosphatidylserine (PS) externalization; upregulated expression of caspase-3, -8, and -9; and apoptotic body formation. In general, the PGGG cell line may serve as a useful tool for studying the toxic mechanisms of cadmium or other toxicants or for toxicity testing and environment monitoring.


Asunto(s)
Apoptosis/efectos de los fármacos , Lubina , Cadmio/toxicidad , Expresión Génica/efectos de los fármacos , Branquias , Proteína 1 Asociada A ECH Tipo Kelch/metabolismo , Contaminantes Químicos del Agua/toxicidad , Animales , Antioxidantes/metabolismo , Catalasa/genética , Catalasa/metabolismo , Técnicas de Cultivo de Célula , Línea Celular , Supervivencia Celular/efectos de los fármacos , Branquias/citología , Proteína 1 Asociada A ECH Tipo Kelch/genética , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/genética , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo
12.
Ecotoxicol Environ Saf ; 209: 111815, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33387774

RESUMEN

Mercury (Hg) is a global contaminant resulting of both natural processes and human activities. In aquatic environments, studies conducted on vertebrates highlighted changes of gene expression or activity of antitoxic and oxidative enzymes. However, although Hg is a highly toxic compound in aquatic environments, only a few studies have evaluated the lethal and sublethal effects of inorganic Hg on Gammarus sp. Therefore, this study aimed at evaluating the effects of inorganic Hg (HgCl2) on the expression of 17 genes involved in crucial biological functions or mechanisms for organisms, namely respiration, osmoregulation, apoptosis, immune and endocrine system, and antioxidative and antitoxic defence systems. The study was performed in males of the freshwater amphipod Gammarus pulex exposed to two environmentally relevant concentrations (50 and 500 ng/L) at two temperature regime fluctuations (16 °C and 20 °C +/-2 °C) for 7 and 21 days. Results showed that G. pulex mortality was dependent on Hg concentration and temperature; the higher the concentration and temperature, the higher the mortality rate. In addition, the Integrated Biomarker Response emphasized that HgCl2 toxicity was dependent on the concentration, time and temperature of exposure. Overall, antioxidant and antitoxic defences, as well as the endocrine and immune systems, were the biological functions most impacted by Hg exposure (based on the concentration, duration, and temperature tested). Conversely, osmoregulation was the least affected biological function. The results also demonstrated a possible adaptation of G. pulex after 21 days at 500 ng/L, regardless of the exposure temperature. This study allowed us to show that Hg deregulates many crucial biological functions after a short exposure, but that during a long exposure, an adaptation phenomenon could occur, regardless of temperature.


Asunto(s)
Anfípodos/fisiología , Expresión Génica/efectos de los fármacos , Mercurio/toxicidad , Temperatura , Contaminantes Químicos del Agua/toxicidad , Anfípodos/metabolismo , Animales , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Agua Dulce , Humanos , Masculino
13.
Carbohydr Polym ; 256: 117514, 2021 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-33483035

RESUMEN

The physicochemical properties, structural features and structure-immunomodulatory activity relationship of pectic polysaccharides from the white asparagus (Asparagus officinalis L.) skin were systematically studied. Using sequential ethanol precipitation, five sub-fractions namely WASP-40, WASP-50, WASP-60, WASP-70 and WASP-80 with distinct degree of esterification (DE) and molecular weight (Mw) were obtained. The Mw and DE values were decreased with the increase of the ethanol concentrations. Structurally, although 4-α-D-GalpA was the dominant sugar residue in all fractions, the molar ratios were decreased, whereas other sugar residues including arabinose- and mannose-based sugar residues overall increased with the increase of ethanol concentration. In addition, the effects of sub-fractions on the RAW 264.7 cells indicated that pectic polysaccharides with the higher DE value showed a stronger immunomodulatory activity. Moreover, the structure-activity relationship was also discussed in this study, which extends the value-added application of asparagus and its processing by-products.


Asunto(s)
Asparagus (Planta)/química , Expresión Génica/efectos de los fármacos , Factores Inmunológicos/química , Fagocitosis/efectos de los fármacos , Polisacáridos/química , Animales , Arabinosa/aislamiento & purificación , Secuencia de Carbohidratos , Proliferación Celular/efectos de los fármacos , Fraccionamiento Químico/métodos , Ésteres/química , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Interleucina-10/genética , Interleucina-10/inmunología , Interleucina-6/genética , Interleucina-6/inmunología , Manosa/aislamiento & purificación , Ratones , Peso Molecular , Extractos Vegetales/química , Polisacáridos/aislamiento & purificación , Polisacáridos/farmacología , Células RAW 264.7 , Relación Estructura-Actividad , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología
14.
Int J Mol Sci ; 22(1)2021 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-33466312

RESUMEN

Despite modern surgical trauma care, bleeding contributes to one-third of trauma-related death. A significant improvement was obtained through the introduction of tranexamic acid (TXA), which today is widely used in emergency and elective orthopedic surgery to control bleeding. However, concerns remain regarding potential adverse effects on bone turnover and regeneration. Therefore, we employed standardized cell culture systems including primary osteoblasts, osteoclasts, and macrophages to evaluate potential effects of TXA on murine bone cells. While osteoblasts derived from calvarial digestion were not affected, TXA increased cell proliferation and matrix mineralization in bone marrow-derived osteoblasts. Short-term TXA treatment (6 h) failed to alter the expression of osteoblast markers; however, long-term TXA stimulation (10 days) was associated with the increased expression of genes involved in osteoblast differentiation and extracellular matrix synthesis. Similarly, whereas short-term TXA treatment did not affect gene expression in terminally differentiated osteoclasts, long-term TXA stimulation resulted in the potent inhibition of osteoclastogenesis. Finally, in bone marrow-derived macrophages activated with LPS, simultaneous TXA treatment led to a reduced expression of inflammatory cytokines and chemokines. Collectively, our study demonstrates a differential action of TXA on bone cells including osteoanabolic, anti-resorptive, and anti-inflammatory effects in vitro which suggests novel treatment applications.


Asunto(s)
Médula Ósea/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Osteoblastos/efectos de los fármacos , Osteoclastos/efectos de los fármacos , Ácido Tranexámico/farmacología , Animales , Médula Ósea/metabolismo , Huesos/efectos de los fármacos , Huesos/metabolismo , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Citocinas/metabolismo , Matriz Extracelular/efectos de los fármacos , Matriz Extracelular/metabolismo , Femenino , Expresión Génica/efectos de los fármacos , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Osteoblastos/metabolismo , Osteoclastos/metabolismo , Osteogénesis/efectos de los fármacos
15.
J Food Sci ; 86(2): 366-375, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33448034

RESUMEN

Liver damage is a common liver disorder, which could induce liver cancer. Oral antioxidant is one of the effective treatments to prevent and alleviate liver damage. In this study, three flavonoids namely myricetin, isoquercitrin, and isorhamnetin were isolated and identified from Laba garlic. The isolated compounds were investigated on the protective effects against H2 O2 -induced oxidative damages in hepatic L02 cells and apoptosis inducing mechanism in hepatic cancer cells HepG2 by using MTT assay, flow cytometry and western blotting analysis. Myricetin, isoquercitrin, and isorhamnetin showed proliferation inhibition on HepG2 cells with IC50 value of 44.32 ± 0.213 µM, 49.68 ± 0.192 µM, and 54.32 ± 0.176 µM, respectively. While they showed low toxicity on normal cell lines L02. They could significantly alleviate the oxidative damage towards L02 cells (P < 0.05), via inhibiting the morphological changes in mitochondria and upholding the integrity of mitochondrial structure and function. The fluorescence intensity of L02 cells pre-treated with myricetin, isoquercitrin, and isorhamnetin (100 µM) was 89.23 ± 1.26%, 89.35 ± 1.43% and 88.97 ± 0.79%, respectively. Moreover, the flavonoids could induce apoptosis in HepG2 cells via Bcl-2/Caspase pathways, where it could up-regulate the expression of Bax and down-regulate the expression of Bcl-2, Bcl-xL, pro-Caspase-3, and pro-Caspase-9 proteins in a dose dependent manner. Overall, the results suggested that the flavonoids from Laba garlic might be a promising candidate for the treatment of various liver disorders. PRACTICAL APPLICATION: Flavonoids from Laba garlic showed selective toxicity towards HepG2 cells in comparison to L02 cells via regulating Bcl-2/caspase pathway. Additionally, the isolated flavonoids expressively barred the oxidative damage induced by H2 O2 in L02 cells. These results suggested that the flavonoids from laba garlic could be a promising agent towards the development of functional foods.


Asunto(s)
Apoptosis/efectos de los fármacos , Caspasas/metabolismo , Flavonoides/farmacología , Ajo/química , Peróxido de Hidrógeno/farmacología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Antineoplásicos Fitogénicos/farmacología , Antioxidantes/farmacología , Línea Celular , Activación Enzimática/efectos de los fármacos , Flavonoides/análisis , Expresión Génica/efectos de los fármacos , Células Hep G2 , Humanos , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Proteínas Proto-Oncogénicas c-bcl-2/genética , Quercetina/análogos & derivados , Quercetina/análisis , Quercetina/farmacología , Transducción de Señal/efectos de los fármacos
16.
Molecules ; 26(3)2021 Jan 22.
Artículo en Inglés | MEDLINE | ID: mdl-33499133

RESUMEN

Low-molecular-weight chitosan (LMWC), a product of chitosan deacetylation, possesses anti-inflammatory effects. In the present study, a porcine small intestinal epithelial cell line, IPEC-J2, was used to assess the protective effects of LMWC on lipopolysaccharide (LPS)-induced intestinal epithelial cell injury. IPEC-J2 cells were pretreated with or without LMWC (400 µg/mL) in the presence or absence of LPS (5 µg/mL) for 6 h. LMWC pretreatment increased (p < 0.05) the occludin abundance and decreased (p < 0.05) the tumour necrosis factor-α (TNF-α) production, apoptosis rate and cleaved cysteinyl aspartate-specific protease-3 (caspase-3) and -8 contents in LPS-treated IPEC-J2 cells. Moreover, LMWC pretreatment downregulated (p < 0.05) the expression levels of TNF receptor 1 (TNFR1) and TNFR-associated death domain and decreased (p < 0.05) the nuclear and cytoplasmic abundance of nuclear factor-κB (NF-κB) p65 in LPS-stimulated IPEC-J2 cells. These results suggest that LMWC exerts a mitigation effect on LPS-induced intestinal epithelial cell damage by suppressing TNFR1-mediated apoptosis and decreasing the production of proinflammatory cytokines via the inhibition of NF-κB signalling pathway.


Asunto(s)
Quitosano/farmacología , Inflamación/prevención & control , Lipopolisacáridos/antagonistas & inhibidores , Lipopolisacáridos/toxicidad , Animales , Apoptosis/efectos de los fármacos , Apoptosis/genética , Caspasas/metabolismo , Línea Celular , Quitosano/administración & dosificación , Quitosano/química , Citocinas/metabolismo , Expresión Génica/efectos de los fármacos , Inflamación/inducido químicamente , Inflamación/metabolismo , Mediadores de Inflamación/metabolismo , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Lipopolisacáridos/administración & dosificación , Peso Molecular , FN-kappa B/metabolismo , Ocludina/metabolismo , Transducción de Señal/efectos de los fármacos , Porcinos
17.
Cytokine ; 140: 155430, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33508651

RESUMEN

In vitro interferon (IFN)α treatment of primary human upper airway basal cells has been shown to drive ACE2 expression, the receptor of SARS-CoV-2. The protease furin is also involved in mediating SARS-CoV-2 and other viral infections, although its association with early IFN response has not been evaluated yet. In order to assess the in vivo relationship between ACE2 and furin expression and the IFN response in nasopharyngeal cells, we first examined ACE2 and furin levels and their correlation with the well-known marker of IFNs' activation, ISG15, in children (n = 59) and adults (n = 48), during respiratory diseases not caused by SARS-CoV-2. A strong positive correlation was found between ACE2 expression, but not of furin, and ISG15 in all patients analyzed. In addition, type I and III IFN stimulation experiments were performed to examine the IFN-mediated activation of ACE2 isoforms (full-length and truncated) and furin in epithelial cell lines. Following all the IFNs treatments, only the truncated ACE2 levels, were upregulated significantly in the A549 and Calu3 cells, in particular by type I IFNs. If confirmed in vivo following IFNs' activation, the induction of the truncated ACE2 isoform only would not enhance the risk of SARS-CoV-2 infection in the respiratory tract.


Asunto(s)
/genética , Células Epiteliales/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Interferones/farmacología , /efectos de los fármacos , Células A549 , Adulto , Antivirales/metabolismo , Antivirales/farmacología , Línea Celular Tumoral , Niño , Citocinas/genética , Células Epiteliales/metabolismo , Humanos , Interferones/metabolismo , Pulmón/citología , Persona de Mediana Edad , Ubiquitinas/genética
19.
J Insect Sci ; 21(1)2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-33400795

RESUMEN

Honey bees are important pollinators of wild plants and crops. MicroRNAs (miRNAs) are endogenous regulators of gene expression. In this study, we initially determined that the lethal concentration 50 (LC50) of dinotefuran was 0.773 mg/l. Then, the expression profiles and differentially expressed miRNAs (DE miRNAs) in honey bee brains after 1, 5, and 10 d of treatment with the lethal concentration 10 (LC10) of dinotefuran were explored via deep small-RNA sequencing and bioinformatics. In total, 2, 23, and 27 DE miRNAs were identified after persistent exposure to the LC10 of dinotefuran for 1, 5, and 10 d, respectively. Some abundant miRNAs, such as ame-miR-375-3p, ame-miR-281-5p, ame-miR-3786-3p, ame-miR-10-5p, and ame-miR-6037-3p, were extremely significantly differentially expressed. Enrichment analysis suggested that the candidate target genes of the DE miRNAs are involved in the regulation of biological processes, cellular processes, and behaviors. These results expand our understanding of the regulatory roles of miRNAs in honey bee Apis mellifera (Hymenopptera: Apidae) responses to neonicotinoid insecticides and facilitate further studies on the functions of miRNAs in honey bees.


Asunto(s)
Encéfalo/efectos de los fármacos , Expresión Génica/efectos de los fármacos , Guanidinas/toxicidad , Insecticidas/toxicidad , MicroARNs/metabolismo , Neonicotinoides/toxicidad , Nitrocompuestos/toxicidad , Animales , Abejas , Encéfalo/metabolismo
20.
Cell Prolif ; 54(2): e12976, 2021 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-33393124

RESUMEN

BACKGROUND: In mammals, early pregnancy is a critical vulnerable period during which complications may arise, including pregnancy failure. Establishment of a maternal endometrial acceptance phenotype is a prerequisite for semiheterogeneous embryo implantation, comprising the rate-limiting step of early pregnancy. METHODS: Confocal fluorescence, immunohistochemistry and western blot for nuclear and cytoplasmic protein were used to examine the activation of yes-associated protein (YAP) in uterine tissue and primary endometrial cells. The target binding between miR16a and YAP was verified by dual-luciferase reporter gene assay. The mouse pregnancy model and pseudopregnancy model were used to investigate the role of YAP in the maternal uterus during early pregnancy in vivo. RESULTS: We showed that YAP translocates into the nucleus in the endometrium of cattle and mice during early pregnancy. Mechanistically, YAP acts as a mediator of ECM rigidity and cell density, which requires the actomyosin cytoskeleton and is partially dependent on the Hippo pathway. Furthermore, we found that the soluble factor IFNτ, which is a ruminant pregnancy recognition factor, also induced activation of YAP by reducing the expression of miR-16a. CONCLUSIONS: This study revealed that activation of YAP is necessary for early pregnancy in bovines because it induced cell proliferation and established an immunosuppressive local environment that allowed conceptus implantation into the uterine epithelium.


Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas de Ciclo Celular/metabolismo , Endometrio/metabolismo , Matriz Extracelular/metabolismo , Interferón Tipo I/metabolismo , Proteínas Gestacionales/metabolismo , Proteínas Adaptadoras Transductoras de Señales/antagonistas & inhibidores , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Antagomirs/metabolismo , Bovinos , Proteínas de Ciclo Celular/antagonistas & inhibidores , Proteínas de Ciclo Celular/genética , Endometrio/citología , Molécula de Adhesión Celular Epitelial/metabolismo , Femenino , Expresión Génica/efectos de los fármacos , Interferón Tipo I/farmacología , Masculino , Ratones , MicroARNs/antagonistas & inhibidores , MicroARNs/genética , MicroARNs/metabolismo , Proteínas Musculares/metabolismo , Embarazo , Proteínas Gestacionales/farmacología , Proteínas Serina-Treonina Quinasas/metabolismo , Interferencia de ARN , ARN Interferente Pequeño/metabolismo , Transducción de Señal , Útero/metabolismo , Útero/patología
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