RESUMO
Vanadium (V) can induce cell apoptosis in layers' oviduct resulting in egg quality reduction. In this study, we investigated the relationship between the mitogen-activated protein kinase (MAPK)-signaling pathway and V-induced apoptosis in poultry oviduct magnum epithelial cells (OMECs). Cultured OMECs were divided into 8 treatment groups: 0 µmol/L V (control), 100 µmol/L V (V100), V100 + P38MAPK inhibitor (SB203580), SB203580, V100 + extracellular signal-regulated kinases 1 and 2 (ERK1/2) inhibitor (U0126), U0126, V100 + c-JUN NH2 -terminal kinase (JNK) inhibitor (SP600125), and SP600125. The OMECs were pretreated with the MAPK inhibitors before their treatment with V100 for 12 h. V100 increased the apoptosis of OMECs (P < .05), while 3 MAPK inhibitors suppressed V100-induced apoptosis P < .05); V100 enhanced the depolarization of â³ψm (P < .05), and SB203580 and U0126 alleviated the V100-induced â³ψm decrease (P < .05); V100 downregulated B-cell lymphoma-2 (Bcl-2) and poly [Adenosine diphosphate ribose] polymerase 1 (PARP1) mRNA expression (P < .05), meanwhile it upregulated Bcl-2 associated x (Bax), Apaf1, cytochrome C (CytC) and cysteine aspartase (caspase) 3, 8, 9 mRNA expression (P < .05). All MAPKs inhibitors alleviated the up-regulation of V100 for Bax and caspase 3 mRNA expression and down-regulation of V100 for Bcl-2 expression (P < .05). SB203580 and U0126 upregulated CytC expression treated by V100 (P < .05), except SP600125, while SB203580 administration resulted in a similar upregulation of PARP1 expression (P < .05). SP600125 can alleviated V triggered p-P38MAPK (phosphor-P38), p-ERK1/2 (phosphor-ERK1/2), p-JNK (phosphor-JNK) increase on OME cells, and SB203580 and U0126 had a similar response to phosphor-P38 and p-JNK (P < .05). It concluded that V-induced apoptosis in OMECs through the activation of P38 and ERK1/2, and by increasing the ratio of Bax/Bcl-2, which resulted in â³ψm decrease, CytC release into the cytosol; consequently caspase 3 is recruited and activated, PARP1 is cleaved, eventually leading to apoptosis.
Assuntos
Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/fisiologia , Mitocôndrias/efeitos dos fármacos , Oviductos/efeitos dos fármacos , Vanádio/toxicidade , Proteínas Quinases p38 Ativadas por Mitógeno/fisiologia , Animais , Células Cultivadas , Galinhas , Células Epiteliais/fisiologia , Feminino , Mitocôndrias/metabolismo , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Oviductos/citologiaRESUMO
Heat stress (HS) is a major environmental threat that affects duck production in subtropical and tropical regions, especially in summer. This study aimed to evaluate the physiological and metabolic responses of Pekin ducks to chronic HS conditions via liquid chromatography-mass spectrometry (LC-MS) using a paired-fed (PF) experimental design. On the basis of equivalent feed intake (HS vs. PF), HS significantly reduced growth performance and the percentage of leg and breast muscles, however, markedly increased the percentage of abdominal fat and breast skin fat. Serum metabolomics results revealed that heat-stressed ducks showed enhanced glycolysis and pentose phosphate pathways, as demonstrated by higher glucose 6-phosphate and 6-phogluconic acid levels in the PF vs. HS comparison. HS decreased hepatic mRNA levels of mitochondrial fatty acid ß-oxidation-related genes (MCAD and SCAD) compared to the PF group, resulting in acetylcarnitine accumulation in serum. Moreover, HS elevated the concentrations of serum amino acids and mRNA levels of ubiquitination-related genes (MuRF1 and MAFbx) in the skeletal muscle and amino acid transporter-related genes (SLC1A1 and SLC7A1) and gluconeogenesis-related genes (PCK1 and PCase) in the liver compared to the PF group. When compared to the normal control group (NC), HS further decreased growth performance, but it elevated the abdominal fat rate. However, increased mRNA levels of ubiquitination-related genes and serum amino acid accumulation were not observed in the HS group compared to the NC group, implying that reduced feed intake masked the effect of HS on skeletal muscle breakdown and is a form of protection for the organism. These results suggest that chronic HS induces protein degradation in the skeletal muscle to provide amino acids for hepatic gluconeogenesis to provide sufficient energy, as Pekin ducks under HS conditions failed to efficiently oxidise fatty acids and ketones in the mitochondria, leading to poor growth performance and slaughter characteristics.
Assuntos
Patos , Resposta ao Choque Térmico , Animais , Fígado/metabolismo , Aminoácidos/metabolismo , RNA Mensageiro/metabolismoRESUMO
Research on the interaction between dietary vitamins and intestinal bacteria is poorly understood. To investigate the effect of dietary vitamins on the cecal bacterial communities, 2 bacterial 16S rRNA gene clone libraries were constructed from pooled PCR products obtained from the cecal digesta of 28-d broilers fed diets with vitamins (V) at the NRC level or with no vitamins (NV). The results showed that BW gain and average feed intake of V broilers was significantly higher (P < 0.01) than NV broilers, whereas the feed/gain ratio was significantly lower (P < 0.01) in V broilers. A total of 188 and 185 clones were sequenced for the NV and V broilers, respectively. Sequence identity criterion of 98% was used to assign sequences to operational taxonomic units (OTU). Clones from the NV group broilers were assigned to 14 OTU, with 33% clones affiliated with the genus Clostridium, 19% affiliated with the genera Escherichia/Shigella, 14% affiliated with the genus Bacteroides, and the remaining clones (34%) affiliated with 5 other bacterial genera (Faecalibacterium, Parasporobacterium, Ruminococcus, Streptococcus, and Subdoligranulum). Clones from the V group broilers were assigned to 23 OTU, with 46% of the clones affiliated with the genus Clostridium, 11% affiliated with the genus Fecalibacterium, and the remaining clones (43%) affiliated with 8 other genera (Anaerofilum, Lactobacillus, Anaerotruncus, Oscillibacter, Alistipes, Gracilibacter, Acetivibrio, and Haloplasma). Three OTU assigned to Clostridium, Faecalibacterium, and Ruminoccus were shared between the 2 libraries. Shannon diversity index showed the V broilers exhibited significantly higher bacterial diversity (P = 0.05), and Libshuff analysis indicated that the community structure between the 2 groups was significantly different (P < 0.0001). These results suggest that lack of dietary vitamins can increase the ratio of facultative pathogenic bacteria and decrease the diversity of bacteria in the cecum of broilers. Our results provide new leads for further investigations on the interaction between dietary vitamin additives and the gut health of broilers.
Assuntos
Bactérias/classificação , Bactérias/genética , Ceco/microbiologia , Galinhas/microbiologia , Galinhas/fisiologia , Vitaminas/metabolismo , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Bactérias/crescimento & desenvolvimento , Bactérias/isolamento & purificação , China , Clonagem Molecular , Suplementos Nutricionais/análise , Biblioteca Gênica , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase/veterinária , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Análise de Sequência de RNA/veterinária , Vitaminas/administração & dosagemRESUMO
In the present study, two experiments were conducted to investigate the effect of Mn source on Mn transport and the expression of a Mn transporter, divalent metal transporter 1 (DMT1), in the small intestine of broilers. In Expt 1, in situ ligated duodenal loops from Mn-deficient chicks (29-d-old) were perfused with solutions containing 0-8.74 mmol Mn/l from either MnSO4, or one of two organic chelates of Mn and amino acids with moderate (OM) or strong (OS) chelation strength (Q(f)) up to 30 min. In Expt 2, Mn-deficient intact broilers (14-d-old) were fed a control diet (12.45 mg Mn/kg) or the control diet supplemented with 100 mg Mn/kg as one of all Mn sources for 14 d. The uptake kinetics of Mn from different Mn sources in the ligated duodenal loops followed a saturable process as determined by regression analysis of concentration-dependent uptake rates. The maximum transport rate (Jmax) and K(m) values, and DMT1 mRNA levels in the ligated duodenal loops were higher (P < 0.01) for OM and OS than for MnSO4. DMT1 mRNA levels were much higher (P < 0.01) in the duodenum than in the jejunum and ileum. Both DMT1 mRNA levels in the duodenum and plasma Mn contents from the hepatic portal vein of intact chicks on day 14 post-feeding increased (P < 0.05) in the following order: control < MnSO4 < OM < OS. These results indicated that organic Mn sources with stronger Q(f) showed higher Mn transport and absorption, and DMT1 might be involved in the regulation of organic Mn transport in the proximal small intestine of broilers.
Assuntos
Proteínas Aviárias/metabolismo , Proteínas de Transporte de Cátions/metabolismo , Galinhas/metabolismo , Regulação da Expressão Gênica , Absorção Intestinal , Intestino Delgado/metabolismo , Manganês/administração & dosagem , Aminoácidos/química , Ração Animal/análise , Animais , Proteínas Aviárias/genética , Proteínas de Transporte de Cátions/genética , Quelantes/administração & dosagem , Quelantes/química , Galinhas/sangue , Galinhas/crescimento & desenvolvimento , China , Deficiências Nutricionais/dietoterapia , Deficiências Nutricionais/metabolismo , Deficiências Nutricionais/veterinária , Duodeno/crescimento & desenvolvimento , Duodeno/metabolismo , Intestino Delgado/crescimento & desenvolvimento , Cinética , Masculino , Manganês/deficiência , Manganês/metabolismo , Manganês/uso terapêutico , Compostos de Manganês/administração & dosagem , Especificidade de Órgãos , Concentração Osmolar , Doenças das Aves Domésticas/dietoterapia , Doenças das Aves Domésticas/metabolismo , Sulfatos/administração & dosagemRESUMO
The objective of this paper was to evaluate the effect of vanadium (V) in high-fat diets sourced from egg yolk on body weight gain, feed intake, blood characteristics and antioxidative status of Wistar rats. A total of 72 female Wistar rats were allocated according to a 2 × 4 factorial design throughout a 5-wk trial, including 2 levels of dietary fat (normal and high; ether extract 40.3 and 301.2 g/kg; fat sourced from egg yolk) and 4 levels of dietary V (0, 3, 15 and 30 mg/kg). Vanadium decreased (P ≤ 0.05) body weight gain (V at 30mg/kg during wk 1 and 2; V at 15 and 30 mg/kg during the overall phase), feed intake (V at 30 mg/kg during wk 3 and the overall phase; V at 15 and 30 mg/kg during wk 4), but increased the relative weight of liver (V at 30 mg/kg, P ≤ 0.05). Moreover, increasing dietary V significantly increased (P ≤ 0.05) plasma aspartate aminotransferase, alanine aminotransferase and malondialdehyde levels and decreased triglyceride level, and V at 30 mg/kg in high-fat treatment had the highest or lowest values (interaction, P ≤ 0.05). Under the same dietary V dose, V residual content in liver (dietary V at 15 and 30 mg/kg) and kidney (dietary V at 15 mg/kg) was higher in high-fat diet treatment compared with normal-fat diet treatment (P ≤ 0.05). In conclusion, it is suggested that V could decrease the body weight together with the feed intake, and the high fat could enhance oxidative stress induced by V of Wistar rats.