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1.
Cell Death Dis ; 10(4): 307, 2019 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-30952838

RESUMO

Di-n-butyl phthalate (DBP) is a kind of ubiquitous chemical linked to hormonal disruptions that affects male reproductive system. However, the mechanism of DBP-induced germ cells toxicity remains unclear. Here, we demonstrate that DBP induces reduction of proliferation, increase of apoptosis and DNA damage dependent on the PTEN/AKT pathway. Mechanistically, DBP decreases PTEN promoter methylation and increases its transcriptional activity, leading to increased PTEN expression. Notably, DNMT3b is confirmed as a target of miR-29b and miR-29b-mediated status of PTEN methylation is involved in the effects of DBP treatment. Meanwhile, DBP decreases AKT pathway expression via increasing PTEN expression. In addition, the fact that DBP decreases the sperm number and the percentage of motile and progressive sperm is associated with downregulated AKT pathway and sperm flagellum-related genes. Collectively, these findings indicate that DBP induces aberrant PTEN demethylation, leading to inhibition of the AKT pathway, which contributes to the reproductive toxicity.

2.
Environ Toxicol ; 32(7): 1908-1917, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28295950

RESUMO

di-N-butylphthalate (DBP) is a ubiquitous environmental pollutant used for plastic coating and in the cosmetics industry. It has toxic effects on body health, especially the male reproductive system. Here, we investigated the effects of DBP on the male reproductive system of pubertal mice and explored the protective role of sulforaphane (SFN). The results showed that DBP significantly reduced the anogenital distance, testicular weight, sperm count and motility, and plasma and testicular testosterone levels and significantly increased the oxidative stress, sperm abnormalities, and testicular cell apoptosis. SFN supplementation ameliorated these effects. After DBP stimulation, the transcription factor nuclear factor erythroid-related factor 2 (Nrf2) was adaptively increased together with its target genes, such as HO-1 and NQO1. Upregulation of Nrf2 by SFN reduced the DBP-mediated intracellular oxidative toxicity and also increased testosterone secretion and spermatogenesis, which were decreased by DBP. These findings indicate that SFN can attenuate DBP-induced reproductive damage in pubertal mice via Nrf2-associated pathways.


Assuntos
Dibutilftalato/toxicidade , Poluentes Ambientais/toxicidade , Isotiocianatos/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Reprodução/efeitos dos fármacos , Animais , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Masculino , Camundongos , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Contagem de Espermatozoides , Motilidade Espermática/efeitos dos fármacos , Testículo/citologia , Testículo/efeitos dos fármacos , Regulação para Cima
3.
Asian J Androl ; 19(4): 404-408, 2017 Jul-Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27080478

RESUMO

Semen cryopreservation is widely used in assisted reproductive technologies, but it reduces sperm quality dramatically. The aim of this study was to develop a model using basal semen quality to predict the outcome of postthaw semen parameters and improve the efficiency of cryopreservation in a human sperm bank. Basal semen parameters of 180 samples were evaluated in the first stage, and a multiple logistic regression analysis involving a backward elimination selection procedure was applied to select independent predictors. After a comprehensive analysis of all results, we developed a new model to assess the freezability of sperm. Progressive motility (PR), straight-line velocity (VSL) and average path velocity (VAP) were included in our model. A greater area under the receiver operating characteristic curve was obtained in our model when compared with other indicators. In the second stage of our study, samples that satisfied the new model were selected to undergo freeze-thawing. Compared with the first stage, the rate of good freezability was increased significantly (94% vs 67%, P = 0.003). By determining basal semen quality, we have developed a new model to improve the efficiency of cryopreservation in a human sperm bank.


Assuntos
Criopreservação/métodos , Preservação do Sêmen/métodos , Bancos de Esperma/métodos , Adulto , Congelamento , Humanos , Masculino , Modelos Estatísticos , Análise Multivariada , Prognóstico , Curva ROC , Análise do Sêmen , Espermatozoides , Adulto Jovem
4.
Nan Fang Yi Ke Da Xue Xue Bao ; 36(9): 1181-1185, 2016 08 20.
Artigo em Chinês | MEDLINE | ID: mdl-27687647

RESUMO

OBJECTIVE: To explore the effect of exposure to vehicle exhaust in pregnant mice on the reproductive function and DNA methylation in male offspring mice. METHODS: Twenty pregnant mice were randomized into control group and vehicle exhaust exposure group (n=10) and exposed to routine laboratory condition and to vehicle exhaust for 10 consecutive days (8 h per day) in a tunnel with a heavy traffic, where the concentrations of TSP, PM10, PM2.5, SO2 and NOX and the decibel of noise were measured. The offspring mice were raised till reaching maturity, and the epididymides of the male mice were collected to test the weight coefficients, DNA methylation level, and mRNA levels of Aldh7a1 and Rpe. RESULTS: The body weight and the weight coefficients of the epididymides and testes differed significantly between the exposure group and the control group (P>0.05). The concentrations of TSP, PM2.5, PM10 and NOx and the decibel of noise were significantly higher in the traffic environment and the control environment (P<0.05). Reduced representation bisulphite sequencing (RRBS) and Gene ontology (GO) showed that 58 genes had significantly different methylation levels between the two groups, mostly relating to the process of spermatogenesis (P<0.05). Compared with the control group, Aldh7a1 and Rpe mRNA expressions in the testes were down-regulated significantly in the exposure group (P<0.05). CONCLUSION: Exposure of pregnant mice to vehicle exhaust causes damages of the reproductive function in the male offspring mice.


Assuntos
Metilação de DNA , Exposição Materna/efeitos adversos , Efeitos Tardios da Exposição Pré-Natal/fisiopatologia , Testículo/fisiopatologia , Emissões de Veículos/toxicidade , Animais , Feminino , Masculino , Camundongos , Gravidez , Espermatogênese/efeitos dos fármacos , Espermatogênese/genética , Testículo/efeitos dos fármacos
5.
Cryobiology ; 71(1): 141-5, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25910678

RESUMO

Sperm cryopreservation is a method to preserve sperm samples for a long period. However, the fertility of sperm decreases markedly after freezing and thawing in a certain amount of samples. The aim of the present study was to find useful and reliable predictive biomarkers of the capacity to withstand the freeze-thawing process in human ejaculates. Previous researches have shown that enolase1 (ENO1) and glucose-6-phosphate isomerase (GPI) are closely related to spermatozoa quality. We chose the two proteins as probable markers of sperm freezing capacity. Ejaculate samples were separated into good freezability ejaculates (GFE) and poor freezability ejaculates (PFE) according to progressive motility of the sperm after thawing. Before starting cryopreservation protocols, the two proteins from each group were compared using western blot analysis and immunofluorescence. Results showed that normalized content of ENO1 (P<0.05) and GPI (P<0.01) were both significantly higher in GFE than in PFE. The association of ENO1 and GPI with postthaw sperm viability and motility was confirmed using Pearson's linear correlation. In conclusion, ENO1 and GPI can be used as markers of human sperm freezability before starting the cryopreservation procedure.


Assuntos
Biomarcadores Tumorais/metabolismo , Criopreservação/métodos , Proteínas de Ligação a DNA/metabolismo , Glucose-6-Fosfato Isomerase/metabolismo , Fosfopiruvato Hidratase/metabolismo , Análise do Sêmen , Preservação do Sêmen/métodos , Espermatozoides/fisiologia , Proteínas Supressoras de Tumor/metabolismo , Biomarcadores/metabolismo , Western Blotting , Sobrevivência Celular , Citocinas/metabolismo , Fertilidade/fisiologia , Congelamento , Humanos , Masculino , Motilidade Espermática/fisiologia , Espermatozoides/enzimologia
6.
Foodborne Pathog Dis ; 6(3): 297-304, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19272004

RESUMO

The aim of the present study was to investigate the antibiotic resistance profiles and the molecular epidemiology of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli isolates from two production swine operations in Sichuan Province, China, between August 2002 and February 2007. The prevalence of ESBL-producing E. coli increased dramatically from 2.2% to 10.7% during this period. This increase appeared mostly related to dissemination of CTX-M-type ESBLs among E. coli isolates. Of 212 E. coli isolates studied, 14 harbored ESBL genes. Among them, 13 harbored bla(CTX-M-15/22) and one harbored bla(SHV-2). To our knowledge, this is the first study to identify bla(CTX-M-22) from production animals. One isolate in 2002 harbored bla(SHV-2), indicating that ESBL genes have been present in farm animals in China since at least 2002. Molecular characterization and pulsed-field gel electrophoresis of the ESBL-producing isolates suggested that different mechanisms may be involved in the dissemination of the CTX-M genes and revealed that additional resistance determinants for non-beta-lactam antibiotics were carried by plasmids encoding certain ESBL genes. Results of this study provide an example of how ESBL genes, particularly those of CTX-M lineages, are rapidly spreading among E. coli isolates from commercial pig farms in Sichuan province of China.


Assuntos
Proteínas de Escherichia coli/análise , Escherichia coli/enzimologia , Fezes/microbiologia , Suínos/microbiologia , beta-Lactamases/análise , Agricultura , Animais , Sequência de Bases , China , Conjugação Genética , DNA Bacteriano/análise , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Escherichia coli/genética , Escherichia coli/isolamento & purificação , Proteínas de Escherichia coli/genética , Plasmídeos/genética , Reação em Cadeia da Polimerase , Resistência beta-Lactâmica/genética , beta-Lactamases/genética
7.
Sheng Wu Gong Cheng Xue Bao ; 22(4): 555-60, 2006 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-16894887

RESUMO

The M protein gene of porcine reproductive and respiratory syndrome virus amplified by PCR was tandem linked with its GP5 gene in shuttle vector in correct frame, resulting in shuttle vector pShuttle-CMV-M-GP5. The positive clone was identified by PCR and further confirmed by sequencing. The constructed plasmid was linearized with Pme I and co-transformed BJ5183 host bacteria with pAdEasy-1 to produce recombinant adenovirus DNA by homologous recombination. Then the adenovirus DNA was linearized with Pac I and transfected into HEK-293A cells to obtain recombinant adenovirus. The specific expression of target proteins by the recombinant adenovirus was verified by indirect immuno-fluorescence assay (IFA) with monoclonal antibodies against M and GP5.The results showed that the tandem linked M with GP5 could be co-expressed by adenovirus vector. Mice immunized with the constructed recombinant adenovirus induced strong humoral immunity (ELISA antibody and virus neutralizing antibody) and cellular immunity (lymphocyte proliferation and CTL responses). The results showed that the recombinant adenovirus has strong immunogenicity and provided the basis for the further experiments in pigs.


Assuntos
Adenoviridae/genética , Proteínas Recombinantes de Fusão/biossíntese , Proteínas do Envelope Viral/genética , Proteínas da Matriz Viral/genética , Animais , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Humanos , Camundongos , Células NIH 3T3 , Plasmídeos , Vírus da Síndrome Respiratória e Reprodutiva Suína/imunologia , Proteínas Recombinantes de Fusão/imunologia , Vacinas Sintéticas/imunologia , Proteínas do Envelope Viral/imunologia , Proteínas da Matriz Viral/imunologia , Vacinas Virais/imunologia
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