RESUMO
BACKGROUND: The B-cell lymphoma 2 (Bcl-2) protein regulates programmed cell death throughout the disease conditions by upholding apoptotic pathways. However, the mechanism by which it's expressed in chondrocytes still needs to be studied in chondrocyte-related disorders. Additionally, exploring the potential therapeutic role of Chlorogenic acid (CGA) in confluence with Bcl-2 modulation is of significant interest. METHODS: In vivo and in vitro studies were performed according to our previous methodologies. The chondrocytes were cultured in specific growth media under standard conditions after expression verification of different microRNAs through high-throughput sequencing and verification of Bcl-2 involvement in tibial growth plates. The effect of Bcl-2 expression was investigated by transfecting chondrocytes with miR-460a, siRNA, and their negative controls alone or in combination with CGA. The RNA was extracted and subjected to a reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Western blot analysis and immunofluorescence assays were performed to visualize the intracellular localization of Bcl-2 and associated proteins related to apoptotic and inflammasome pathways. Moreover, apoptosis through flow cytometry was also performed to understand the modulation of concerning pathways. RESULTS: The suppression of Bcl-2 induced higher apoptosis and mitochondrial dysfunction, leading to IL-1ß maturation and affecting the inflammasome during chondrocyte proliferation. Conversely, overexpression attenuated the activation, as evidenced by reduced caspase activity and IL-1ß maturation. In parallel, CGA successfully reduced siRNA-induced apoptosis by decreasing Cytochrome C (Cyto C) release from the mitochondria to the cytoplasm, which in turn decreased Caspase-3 and Caspase-7 cleavage with Bcl-2-associated X protein (Bax). Furthermore, siBcl-2 transfection and CGA therapy increased chondrocyte proliferation and survival. The CGA also showed a promising approach to maintaining chondrocyte viability by inhibiting siRNA-induced apoptosis. CONCLUSIONS: Targeting Bcl-2-mediated regulation might be a possible treatment for chondrocyte-related conditions. Moreover, these results add knowledge of the complicated processes underlying chondrocyte function and the pathophysiology of related diseases, highlighting the significance of target specific therapies. Video Abstract.
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Condrócitos , MicroRNAs , Condrócitos/metabolismo , Inflamassomos/metabolismo , Ácido Clorogênico/farmacologia , Ácido Clorogênico/metabolismo , Apoptose , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Interferente Pequeno/metabolismo , Interleucina-1beta/metabolismoRESUMO
Aflatoxin B1 (AFB1) and zearalenone (ZEN) cause serious damage to mammals, but few studies have investigated the impacts of these toxins on pregnant and lactating mammals. This study investigated the effects of ZEN on AFB1-induced intestinal and ovarian toxicity in pregnant and lactating rats. Based on the results, AFB1 reduces the digestion, absorption, and antioxidant capacity in the intestine, increases intestinal mucosal permeability, destroys intestinal mechanical barriers, and increases pathogenic bacteria' relative abundances. Simultaneously, ZEN can exacerbate the intestinal injury caused by AFB1. The intestines of the offspring were also damaged, but the damage was less severe than that observed for the dams. While AFB1 activates various signalling pathways in the ovary and affects genes related to endoplasmic reticulum stress, apoptosis, and inflammation, ZEN may exacerbate or antagonize the AFB1 toxicity on gene expression in the ovary through key node genes and abnormally expressed genes. Our study found that mycotoxins can not only directly damage the ovaries and affect gene expression in the ovaries but can also impact ovarian health by disrupting intestinal microbes. Mycotoxins are an important environmental pathogenic factor for intestinal and ovarian disease in pregnancy and lactation mammals.
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Micotoxinas , Tricotecenos , Zearalenona , Animais , Ratos , Gravidez , Feminino , Zearalenona/toxicidade , Tricotecenos/toxicidade , Aflatoxina B1/toxicidade , Ovário , Lactação , Intestinos , MamíferosRESUMO
Chronic wound is a common complication for diabetic patients, which entails substantial inconvenience, persistent pain, and significant economic burden to patients. However, current clinical treatments for diabetic chronic wounds remain unsatisfactory. A prolonged but ineffective inflammation phase in chronic wounds is the primary difference between diabetic chronic wounds and normal wounds. Herein, we present an effective antioxidative system (MOF/Gel) for chronic wound healing of diabetic rats through integrating a metal organic framework (MOF) nanozyme with antioxidant enzyme-like activity with a hydrogel (Gel). MOF/Gel can continuously scavenge reactive oxygen species to modulate the oxidative stress microenvironment in diabetic chronic wounds, which leads to a natural transition from the inflammation phase to the proliferation phase. Impressively, the efficacy of one-time-applied MOF/Gel was comparable to that of the human epidermal growth factor Gel, a widely used clinical drug for various wound treatments. Such an effective, safe, and convenient MOF/Gel system can meet complex clinical demands.
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Diabetes Mellitus Experimental , Estruturas Metalorgânicas , Nanopartículas , Humanos , Ratos , Animais , Estruturas Metalorgânicas/farmacologia , Diabetes Mellitus Experimental/tratamento farmacológico , Cicatrização , Antioxidantes/farmacologia , Nanopartículas/uso terapêutico , Inflamação , Hidrogéis/farmacologiaRESUMO
Food and feed are frequently co-contaminated with aflatoxin B1 (AFB1) and zearalenone (ZEN). This study investigated the effects of ZEN on the AFB1-induced liver and mammary gland toxicity in pregnant and lactating rats. AFB1 and ZEN co-exposure inhibited the growth of rats and caused oxidative stress and inflammatory responses in the liver and mammary gland. Compared with the AFB1-only group, damage was aggravated in the AFB1 + 10 mg/kg ZEN group, and the AFB1 + 1 mg/kg ZEN group showed a reduction in some metrics. The metabolomic results of the mammary gland showed that metabolite changes were mainly in lipid, amino acid, and glucose metabolism. Compared with the AFB1 + 0 mg/kg ZEN group, the AFB1 + 1 mg/kg ZEN group had the most metabolite changes. Moreover, AFB1 and ZEN co-exposure reduced the levels of sex hormones and RNA m6A methylation in the mammary gland. We speculate that ZEN affects the toxicity of AFB1 to the liver and mammary gland by interfering with the function of sex hormones, regulating cell proliferation and metabolic processes.
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Zearalenona , Aflatoxina B1/toxicidade , Aminoácidos , Animais , Feminino , Glucose , Lactação , Lipídeos , Fígado , Gravidez , RNA , Ratos , Zearalenona/toxicidadeRESUMO
Aflatoxin is a known mycotoxin that pollutes various grains widely in the environment. Aflatoxin B1 (AFB1) and Aflatoxin M1 (AFM1) have been shown to induce cytotoxicity in many cells, yet their effects on mammary epithelial cells remain unclear. In this study, we examined the toxicity and the effects of AFB1 and AFM1 on bovine mammary epithelial cells (BME cells). The cells were treated with AFB1 or AFM1 at a concentration of 0-10 mg/L for 24 or 48 h, followed by cytotoxicity assays, flow cytometry, and transcriptomics. Our results demonstrated that AFB1 and AFM1 induced cell proliferation inhibition, apoptosis and cell cycle arrest. However, the level of intracellular reactive oxygen species has no significant difference. The RNA-Seq results also showed that AFB1 and AFM1 changed many related gene expressions like apoptosis and oxidative stress, cycle, junction, and signaling pathway. Taken together, AFB1 and AFM1 were found to affect cytotoxicity and related gene changes in BME cells. Notably, this study reported that 2 mg/L of AFB1 and AFM1 affected the expression of methylation-related genes, and ultimately altered the rate of m6A methylation in RNA. It may provide a potential direction for toxins to indirectly regulate gene expression by affecting RNA methylation modification. Our research provides some novel insights and data about AFB1 and AFM1 toxicity in BME cells.
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Aflatoxina B1/toxicidade , Aflatoxina M1/toxicidade , Testes de Toxicidade , Transcriptoma/fisiologia , Animais , Apoptose/efeitos dos fármacos , Bovinos , Contagem de Células , Proliferação de Células , Células Epiteliais/efeitos dos fármacos , Feminino , Citometria de Fluxo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de OxigênioRESUMO
This study demonstrates the effects of progesterone on eggshell quality and ultrastructure by injecting progesterone into laying hens 2 and 5 h post-oviposition, respectively. Progesterone injected 2 h post-oviposition (P4-2 h) improved eggshell quality with a significant decrease (P < 0.01) in the thickness of the mammillary layer and a significant increase (P < 0.01) in the thickness of the effective layer in the eggshell ultrastructure compared to the control. Progesterone injected 5 h post-oviposition (P4-5 h) damaged the eggshell quality by significantly reducing (P < 0.01) the effective layer thickness. Progesterone injected delayed obviously (P < 0.01) the following oviposition. Moreover, the concentrations of Thr, Cys, Leu, Lys, and His in the eggshell membranes were significantly higher (P < 0.05) in the P4-2 h treated hens whereas Val and Lys were significantly lower (P < 0.05) in P4-5 h treated hens compared to the control. Therefore, progesterone shows paradoxical effects on eggshell quality depending on the injection time-points post-oviposition, which could explain the contradictions in previous related reports. P4 injected affected the content of amino acids in eggshell membranes, especially lysine which contributed to eggshell quality. In addition, P4 injected 2 h after oviposition improved eggshell quality by promoting the premature fusion of mammillary knobs. This work contributed to a novel insight to understanding the mechanism of improving eggshell quality.
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Casca de Ovo/efeitos dos fármacos , Oviposição/efeitos dos fármacos , Progesterona/farmacologia , Animais , Galinhas/genética , Casca de Ovo/química , Feminino , Oviposição/genéticaRESUMO
BACKGROUND: Eggshell breaking strength is critical to reduce egg breaking rate and avoid economic loss. The process of eggshell calcification initiates with the egg entering the uterus and lasts about 18 h. It follows a temporal sequence corresponding to the initiation, growth and termination periods of shell calcification. During each period of shell calcification, our study investigated the differences of gonadal hormones and uterine transcriptome in laying hens producing a high or low breaking strength shell. RESULTS: 60 Hy-line Brown laying hens were selected and divided into two groups according to eggshell breaking strength. Eggshell breaking strength of 44.57 ± 0.91 N and 26.68 ± 0.38 N were considered to be the high strength group (HS) and low strength group (LS), respectively. The results showed that mammillary thickness and mammillary knob width of eggshells were significantly lower in the HS. Serum progesterone (P4) and 1,25-dihydroxy vitamin D3 [1,25-(OH)2D3] were significantly higher in the HS compared to the LS during the initiation period of calcification. Serum estradiol (E2) and calcium did not change significantly. All factors mentioned above had no significant differences in the growth and termination periods of calcification. The relative expression of CaBP-D28k and PMCA 1b were not significantly different between HS and LS. The relative expression of NCX1 was significantly higher in HS compared to LS. Moreover, 1777 differentially expressed genes (DEGs) were obtained in the initiation period of calcification. However, few DEGs were identified in the growth or termination periods of calcification. 30 DEGs were selected as candidate genes involved in eggshell calcification during the initiation period of calcification by the analysis of GO terms and KEGG pathways. CONCLUSIONS: Our study concluded that mammillary thickness and mammillary knob width of the HS were significantly lower than LS. P4 and 1,25-(OH)2D3 were significantly higher in the initiation period of HS. They may impact initial calcification when the mammillary layer is formed. The initiation period of calcification determined eggshell strength rather than the growth or termination periods. We inferred P4 or 1,25-(OH)2D3 may effect the ultrastructure of the mammillary layer by regulating the expression of uterine genes.
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Calcificação Fisiológica/genética , Casca de Ovo/fisiologia , Hormônios Gonadais/sangue , Fenômenos Mecânicos , Oviposição/genética , Transcriptoma , Útero/metabolismo , Animais , Fenômenos Biomecânicos , Cálcio/sangue , Galinhas , Duodeno/metabolismo , FemininoRESUMO
BACKGROUND: Selenium (Se) plays a protective role in aflatoxin B1 (AFB1)-induced splenic immunotoxicity in chicks. OBJECTIVE: This study was designed to reveal the underlying mechanism of Se-mediated protection against AFB1-induced splenic injury in broilers. METHODS: Four groups of 1-d-old Cobb male broilers (n = 5 cages/diet, 6 chicks/cage) were arranged in a 3-wk 2 × 2 factorial design trial whereby they were fed an Se-deficient, corn- and soy-based diet [base diet (BD), 36 µg Se/kg], BD plus 1.0 mg AFB1/kg, BD plus 0.3 mg Se/kg, or BD plus 1.0 mg AFB1/kg and 0.3 mg Se/kg (as 2-hydroxy-4-methylselenobutanoic acid). Serum and spleen were collected at week 3 to assay for cytokines, histology, redox status, selected inflammation- and apoptosis-related genes and proteins, and the selenogenome. RESULTS: Dietary AFB1 induced growth retardation and spleen injury, decreasing (P < 0.05) body weight gain, feed intake, feed conversion efficiency, and serum interleukin-1ß by 17.8-98.1% and increasing (P < 0.05) the spleen index and serum interleukin-6 by 37.6-113%. It also reduced the splenic lymphocyte number, the white pulp region, and histiocyte proliferation in Se-adequate groups. However, Se deficiency aggravated (P < 0.05) these AFB1-induced alterations by 16.2-103%. Moreover, Se deficiency decreased (P < 0.05) splenic glutathione peroxidase (GPX) activity and glutathione-S transferase and glutathione concentrations by 35.6-89.4% in AFB1-exposed groups. Furthermore, Se deficiency upregulated (P < 0.05) the apoptotic (Caspase 3 and Caspase 9) and antimicrobial (ß defensin 1 and 2) genes, but downregulated (P < 0.05) antiapoptotic (B-cell lymphoma 2) and inflammatory (E3 ubiquitin-protein ligase CBL-B) genes at the mRNA and/or protein level in AFB1 supplementation groups. Additionally, Se deficiency downregulated (P < 0.05) GPX3, thioredoxin reductase 1 (TXNRD 1), GPX4, and selenoprotein (SELENO) S, and upregulated (P < 0.05) SELENOT and SELENOU in spleen in AFB1 administered groups. CONCLUSIONS: Dietary Se deficiency exacerbated AFB1-induced spleen injury in chicks, partially through the regulation of oxidative stress, inflammatory and apoptotic signaling, and 6 selenoproteins.
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Aflatoxina B1/toxicidade , Proteínas Aviárias/genética , Selênio/deficiência , Selenoproteínas/genética , Baço/efeitos dos fármacos , Baço/imunologia , Animais , Animais Recém-Nascidos , Antioxidantes/metabolismo , Apoptose/efeitos dos fármacos , Apoptose/genética , Apoptose/imunologia , Galinhas , Regulação da Expressão Gênica/efeitos dos fármacos , Inflamação/etiologia , Inflamação/genética , Inflamação/imunologia , Masculino , Oxirredução , Transdução de Sinais/efeitos dos fármacos , Baço/metabolismoRESUMO
Background: Zearalenone (ZEN) can cause serious defects in development and reproduction in humans and animals. Silymarin shows antioxidant and estrogenic effects. Objective: This study was conducted to determine if silymarin can antagonize ZEN-induced hepatic and reproductive toxicities. Methods: Thirty-five 21-d-old female Sprague-Dawley rats (n = 7/diet) were fed a control diet (Ctrl) or Ctrl plus 20 mg ZEN/kg or Ctrl plus 20 mg ZEN/kg with 100, 200, or 500 mg silymarin/kg for 6 wk. Serum, livers, ovaries, and uterus were collected at week 6 for biochemistry, hormone, and redox status and selected gene and protein assays. Results: The consumption of ZEN decreased (P < 0.05) the final body weight by 17.9%, induced liver injury, increased (P < 0.05) aspartate aminotransferase and alkaline phosphatase activities, and decreased (P < 0.05) total protein and albumin concentrations in serum by 16.7-40.6%. ZEN also caused reproductive toxicity, including decreased (P < 0.05) 17ß-estradiol and increased (P < 0.05) follicle-stimulating hormone concentrations in serum by 12.7-46.3% and induced histopathologic alterations in the liver, ovaries, and uterus. Interestingly, these alterations induced by ZEN were alleviated (P < 0.05) by silymarin supplementation at 100, 200, and 500 mg/kg. Moreover, silymarin supplementation at the 3 doses mitigated (P < 0.05) ZEN-induced impairment in hepatic glutathione peroxidase activity, total antioxidant capacity, and malondialdehyde concentration by 17.6-100%. Meanwhile, silymarin supplementation at all doses upregulated (P < 0.05) phospho-ribosomal protein S6 kinase 1 (p-RPS6KB1) and 3ß-hydroxysteroid dehydrogenase (HSD3B) by 43.0-121% but downregulated (P < 0.05) AMP-activated protein kinase (AMPK) and 3α-hydroxysteroid dehydrogenase (HSD3A) in the liver relative to the ZEN group by 11.2-40.6%. In addition, silymarin supplementation at all doses elevated (P < 0.05) HSD3B by 1.8- to 2.5-fold and decreased (P < 0.05) estrogen receptor 1 (ESR1), ATP binding cassette (ABC) c1, and Abcc5 in ovaries and the uterus by 10.7-63.2%. Conclusion: Dietary silymarin supplementation at 100, 200, and 500 mg/kg protected rats from ZEN-induced hepatotoxicity and reproductive toxicity, potentially through improvement in the antioxidant capacity and regulation in the genes related to protein synthesis, ZEN metabolism, hormone synthesis, and ABC transporters in the tissues.
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Antioxidantes/uso terapêutico , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Fígado/efeitos dos fármacos , Reprodução/efeitos dos fármacos , Silybum marianum/química , Silimarina/uso terapêutico , Zearalenona/toxicidade , Proteínas Quinases Ativadas por AMP/metabolismo , Transportadores de Cassetes de Ligação de ATP/metabolismo , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Proteínas Sanguíneas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/patologia , Suplementos Nutricionais , Receptor alfa de Estrogênio/sangue , Feminino , Glutationa Peroxidase/metabolismo , Hormônios/sangue , Hidroxiesteroide Desidrogenases/metabolismo , Fígado/enzimologia , Fígado/patologia , Malondialdeído/sangue , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Ovário/efeitos dos fármacos , Ovário/patologia , Fitoterapia , Ratos Sprague-Dawley , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , Silimarina/farmacologia , Útero/efeitos dos fármacos , Útero/patologiaRESUMO
Background: A new organic selenium compound, 2-hydroxy-4-methylselenobutanoic acid (SeO), displayed a greater bioavailability than sodium selenite (SeNa) or seleno-yeast (SeY) in several species.Objective: This study sought to determine the regulation of the speciation of selenium, expression of selenogenome and selenocysteine biosynthesis and degradation-related genes, and production of selenoproteins by the 3 forms of selenium in the tissues of broiler chicks.Methods: Day-old male chicks (n = 6 cages/diet, 6 chicks/cage) were fed a selenium-deficient, corn and soy-based diet [base diet (BD), 0.05 mg Se/kg] or the BD + SeNa, SeY, or SeO at 0.2 mg Se/kg for 6 wk. Plasma, livers, and pectoral and thigh muscles were collected at weeks 3 and 6 to assay for total selenium, selenomethionine, selenocysteine, redox status, and selected genes, proteins, and enzymes.Results: Although both SeY and SeO produced greater concentrations (P < 0.05) of total selenium (20-172%) and of selenomethionine (≤15-fold) in the liver, pectoral muscle, and thigh than those of SeNa, SeO further raised (P < 0.05) these concentrations by 13-37% and 43-87%, respectively, compared with SeY. Compared with the BD, only SeO enhanced (P < 0.05) the mRNA of selenoprotein (Seleno) s and methionine sulfoxide reductase B1 (Msrb1) in the liver and thigh (62-98%) and thioredoxin reductase (TXRND) activity in the pectoral and thigh muscles (20-37%) at week 3. Furthermore, SeO increased (P < 0.05) the expression of glutathione peroxidase (Gpx) 3, GPX4, SELENOP, and SELENOU relative to the SeNa group by 26-207%, and the expression of Selenop, O-phosphoseryl-transfer RNA (tRNA):selenocysteinyl-tRNA synthase, GPX4, and SELENOP relative to the SeY group by 23-55% in various tissues.Conclusions: Compared with SeNa or SeY, SeO demonstrated a unique ability to enrich selenomethionine and total selenium depositions, to induce the early expression of Selenos and Mrsb1 mRNA and TXRND activity, and to enhance the protein production of GPX4, SELENOP, and SELENOU in the tissues of chicks.
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Butiratos/farmacologia , Fígado/efeitos dos fármacos , Músculos/efeitos dos fármacos , Compostos de Selênio/farmacologia , Selênio/metabolismo , Selenometionina/metabolismo , Selenoproteínas/metabolismo , Aminoacil-tRNA Sintetases/metabolismo , Fenômenos Fisiológicos da Nutrição Animal , Animais , Butiratos/metabolismo , Galinhas , Glutationa Peroxidase/metabolismo , Fígado/metabolismo , Masculino , Metionina Sulfóxido Redutases/genética , Metionina Sulfóxido Redutases/metabolismo , Músculos/metabolismo , RNA Mensageiro/metabolismo , Selênio/deficiência , Compostos de Selênio/metabolismo , Selenoproteínas/genética , Selenito de Sódio/farmacologia , Tiorredoxina Dissulfeto Redutase/metabolismo , LevedurasRESUMO
There are two classic adipose tissues in mammals, white adipose tissue (WAT) and brown adipose tissue (BAT). It has been well known that browning of WAT can be induced by cold exposure. In this study, to identify the novel cold responsive key miRNAs that are involved in browning, mice were housed at 6 °C for 10 days, and deep sequencing of the miRNAs of WAT and BAT was performed. Our data showed that WAT and BAT displayed distinct expression profiles due to their different locations, morphology and biological function. A total of 27 BAT and 29 WAT differentially expressed (DE) miRNAs were identified in response to cold stimulation, respectively (fold change >2 and false discovery rate (FDR) <0.05), of which, 9 were overlapped in both adipose tissues. Furthermore, the potential target genes of the DE miRNAs from BAT and WAT were predicted computationally, and the KEGG pathway analysis revealed the enrichment pathways in cold stimulated adipose tissues. The expression pattern of miR-144-3p/Bmpr1b/Phlda1 and miR-146a-5p/Sphk2 were further measured by qPCR. Finally, we found that miR-146a-5p was significantly induced during the primary adipogenesis caused by BAT differentiation, whereas miR-144-3p was decreased. Our study identifies for the first time the novel miRNAs involved in browning of WAT by sequencing and expands the therapeutic approaches for combating metabolic diseases.
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Tecido Adiposo Marrom/metabolismo , Tecido Adiposo Branco/metabolismo , Regulação da Expressão Gênica , MicroRNAs/genética , Adipogenia , Tecido Adiposo Marrom/citologia , Tecido Adiposo Branco/citologia , Animais , Células Cultivadas , Temperatura Baixa , Perfilação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BLRESUMO
BACKGROUND: The involvement of cytochrome P450 (CYP450) isozymes and the selenogenome in selenium-mediated protection against aflatoxin B1 (AFB1)-induced adverse effects in broilers remains unclear. OBJECTIVE: This study was designed first to determine whether selenium could reduce AFB1-induced hepatotoxic effects and then to determine whether these effects were due to changes in the CYP450 isozymes and selenogenome expression in the liver of chicks. METHODS: Male avian broilers (aged 120 d) were allocated to 4 groups with 5 replicates of 6 birds to be included in a 2-by-2 factorial trial in which the main factors included supplementation of AFB1 (<5 compared with 100 µg/kg) and selenium (0.2 compared with 0.5 mg/kg) in a corn/soybean-based diet for 4 wk. Serum biochemistry, hepatic histology, and mRNA and/or activities of hepatic antioxidant enzymes, CYP450 isozymes, and 26 selenoproteins were analyzed at week 2 and/or 4. RESULTS: Administration of AFB1 induced liver injury, decreasing (P < 0.05) total protein and albumin concentrations by 33.3-43.8% and increasing (P < 0.05) alanine aminotransferase and aspartate aminotransferase activities by 26.0-33.8% in serum, and induced hepatic necrosis and bile duct hyperplasia at week 2. AFB1 also decreased (P < 0.05) hepatic activities of glutathione peroxidase (GPX), thioredoxin reductase (TXNRD), and catalase, and the glutathione concentration by 13.1-59.9% and increased (P < 0.05) malondialdehyde, 8-hydroxydeoxyguanosine and exo-AFB1-8,9-epoxide (AFBO) DNA concentrations by 17.9-1200%. In addition, the mRNA and activity of enzymes responsible for the bioactivation of AFB1 into AFBO, which included CYP450 A1, 1A2, 2A6, and 3A4, were significantly induced (P < 0.05) by 29.2-271% in liver microsomes after 2-wk exposure to AFB1. These alterations induced by AFB1 were prevented by selenium supplementation. Dietary selenium supplementation increased (P < 0.05) mRNA and/or activities of 6 selenoprotein genes (Gpx3, Txnrd1, Txnrd2, Txnrd3, iodothyronine deiodinase 2, and selenoprotein N) in the liver of AFB1-treated groups at week 2. CONCLUSIONS: Dietary selenium protected chicks from AFB1-induced liver injury, potentially through the synergistic actions of inhibition of the pivotal CYP450 isozyme-mediated activation of AFB1 to toxic AFBO, and increased antioxidant capacities by upregulation of selenoprotein genes coding for antioxidant proteins.
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Aspergillus flavus conidia are widespread in air; they attach to food and feed crops and secrete aflatoxins, which results in serious contamination. Germination of A. flavus conidia is the most critical step in contamination of food by A. flavus. This study aims to gain an insight into A. flavus conidia through dormancy to germination to provide a theoretical basis for inhibition of A. flavus conidia germination. The morphological changes and regulation mechanism of A. flavus conidia germination at 0, 4, 8, and 12 hours were observed. Transcriptomic and metabolomic analyses showed that conidia became active from dormancy (0 hour) to the initial stage of germination (4 hours), cellular respiration and energy metabolism increased, and amino acids and lipids were synthesized rapidly. The number of differentially expressed genes and differential metabolites was highest at this stage. Besides, we found that conidia germination had selectivity for different carbon and nitrogen sources. Compared with monosaccharides, disaccharides, as the only carbon source, significantly promoted the germination of conidia. Moreover, MepA, one of genes in the ammonium transporter family was studied. The gene deletion mutant ΔMepA had a significant growth defect, and the expression of MeaA was significantly upregulated in ΔMepA compared with the wild-type, indicating that both MepA and MeaA played an important role in transporting ammonium ions.IMPORTANCEThis is the first study to use combined transcriptomic and metabolomics analyses to explore the biological changes during germination of Aspergillus flavus conidia. The biological process with the highest changes occurred in 0-4 hours at the initial stage of germination. Compared with polysaccharides, monosaccharides significantly increased the size of conidia, while significantly decreasing the germination rate of conidia. Both MeaA and MepA were involved in ammonia transport and metabolism during conidia germination.
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The purpose of this study was to explore the effects of Res and EGCG on cell growth, cellular antioxidant levels, and cellular lipid metabolism in hepatocytes. In this experiment, leghorn male hepatoma (LMH) cells were used as hepatocytes. The results showed that 6.25-25 µM Res and EGCG had no adverse effects on cell viability and growth. Meanwhile, with the increasing dosage of Res and EGCG, the contents of total cholesterol (TC), total glyceride (TG), and malondialdehyde (MDA) in hepatocytes decreased significantly (p < 0.05), while the contents of glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD), and catalase (CAT) increased significantly (p < 0.05). In addition, western blot results showed that Res and EGCG could significantly increase the expression of p-AMPK protein and reduce the expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) protein in hepatocytes (p < 0.05). Moreover, q-PCR results showed that with the increase in Res and EGCG, the expression of cholesterol- and fatty acid synthesis-related genes decreased significantly (p < 0.05). In conclusion, Res and EGCG can increase the antioxidant capacity of hepatocytes and reduce the synthesis of TC and TG in hepatocytes by activating AMPK, thereby regulating lipid metabolism in hepatocytes.
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The aim of this study was to investigate the effects of aflatoxin B1 (AFB1) on cholestasis in duck liver and its nutritional regulation. Three hundred sixty 1-day-old ducks were randomly divided into six groups and fed for 4 weeks. The control group was fed a basic diet, while the experimental group diet contained 90 µg/kg of AFB1. Cholestyramine, atorvastatin calcium, taurine, and emodin were added to the diets of four experimental groups. The results show that in the AFB1 group, the growth properties, total bile acid (TBA) serum levels and total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and glutathione (GSH) liver levels decreased, while the malondialdehyde (MDA) and TBA liver levels increased (p < 0.05). Moreover, AFB1 caused cholestasis. Cholestyramine, atorvastatin calcium, taurine, and emodin could reduce the TBA serum and liver levels (p < 0.05), alleviating the symptoms of cholestasis. The qPCR results show that AFB1 upregulated cytochrome P450 family 7 subfamily A member 1 (CYP7A1) and cytochrome P450 family 8 subfamily B member 1 (CYP8B1) gene expression and downregulated ATP binding cassette subfamily B member 11 (BSEP) gene expression in the liver, and taurine and emodin downregulated CYP7A1 and CYP8B1 gene expression (p < 0.05). In summary, AFB1 negatively affects health and alters the expression of genes related to liver bile acid metabolism, leading to cholestasis. Cholestyramine, atorvastatin calcium, taurine, and emodin can alleviate AFB1-induced cholestasis.
Assuntos
Aflatoxina B1 , Colestase , Patos , Fígado , Animais , Aflatoxina B1/toxicidade , Colestase/induzido quimicamente , Colestase/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Ácidos e Sais Biliares/metabolismo , Ácidos e Sais Biliares/sangue , Doenças das Aves Domésticas/induzido quimicamente , Resina de Colestiramina/farmacologia , Ração AnimalRESUMO
The growing period is a critical period for the growth and development of hens and affects their production performance during the laying period. During the early stage of growing, bone and muscle growth is rapid, making it necessary to provide sufficient amino acids (AA) to support the growth and development of laying hens. In this experiment, RNA-Sequencing (RNA-Seq) was applied to compare the liver tissues from 6- to 12-wk-old growing laying hens to identify candidate genes related to growth and AA transport and metabolism. In the liver tissues, 596 differentially expressed genes (DEG) were identified, of which 424 genes were up-regulated and 172 were down-regulated. Through enrichment analysis and DEGs analysis, some DEGs and pathways related to AA transport and metabolism were identified. Additionally, there were significantly increased activities in the liver of glutamate dehydrogenase (GDH), glutamic oxaloacetic transaminase (GOT), and glutamate pyruvate transaminase (GPT). Meanwhile, the level of serum insulin-like growth factor binding protein-5 (IGFBP-5) significantly elevated, and insulin-like growth factor-1 (IGF-1) levels significantly reduced at 12 wk compared to 6 wk. The AA contents in the breast muscle were not significantly altered, while the levels of the free AA in the serum underwent significant changes. This study discovered that the transport and metabolism of AAs in growing laying hens at different ages changed, which influenced the growth and development of growing laying hens. This contributes to future research on the mechanisms of growth and AA metabolism during the growing period of laying hens.
Assuntos
Galinhas , Transcriptoma , Animais , Feminino , Galinhas/fisiologia , Fígado/metabolismo , Aminoácidos/metabolismoRESUMO
Global climate change has altered the frequency of soil freeze-thaw cycles, but the response of soil microorganisms to different elevation gradients during the early freeze-thaw period remains unclear. So far, the influence of the altitudinal gradient on the microbial community and metabolic characteristics in the early freeze-thaw period of the Qinghai Lake Basin remains unclear. To this end, we collected soil at different elevations in the early freeze-thaw period of the Qinghai Lake Basin and investigated the influence of the elevation gradient on soil microbial community characteristics and soil metabolic processes as well as the corresponding environmental driving mechanism by high-throughput sequencing and LC-MS (Liquid Chromatograph-Mass Spectrometer) nontargeted metabolite determination. The results showed that Proteobacteria were the dominant microflora in the Qinghai Lake Basin. The dominant phyla associated with carbon and nitrogen are Proteobacteria and Firmicutes, both of which are significantly affected by elevation. The soil physicochemical factors jointly affected the soil microbial communities and metabolism. Total phosphorus nitrate nitrogen and pH were the main driving factors of the microbial community, and metabolites were sensitive to changes in chemical factors. In short, the microbial community structure and function, soil physicochemical factors and soil metabolic processes were significantly affected by the altitudinal gradient in the early freeze-thaw period, while the microbial community diversity showed no significant response to the altitudinal gradient. Additionally, a high potassium content in the soil may promote the growth and reproduction of bacteria associated with carbon and nitrogen cycling, as well as the production of metabolites.
RESUMO
Cell death is a universal biological process in almost every physiological and pathological condition, including development, degeneration, inflammation, and cancer. In addition to apoptosis, increasing numbers of cell death types have been discovered in recent years. The biological significance of cell death has long been a subject of interest and exploration and meaningful discoveries continue to be made. Ferroptosis is a newfound form of programmed cell death and has been implicated intensively in various pathological conditions and cancer therapy. A few studies show that ferroptosis has the direct capacity to kill cancer cells and has a potential antitumor effect. As the rising role of immune cells function in the tumor microenvironment (TME), ferroptosis may have additional impact on the immune cells, though this remains unclear. In this study we focus on the ferroptosis molecular network and the ferroptosis-mediated immune response, mainly in the TME, and put forward novel insights and directions for cancer research in the near future.
Assuntos
Ferroptose , Neoplasias , Humanos , Apoptose , Morte Celular , Inflamação , Microambiente TumoralRESUMO
Sepsis-associated acute kidney injury is a common complication of sepsis, but it is difficult to predict sepsis-associated acute kidney injury. In this retrospective observational study, adult septic patients were recruited from the MIMIC-III database as the training cohort (n = 4764) and from Xiangya Hospital (n = 1568) and Zhang's database as validation cohorts. We identified eleven predictors with seven independent risk predictors of sepsis-associated acute kidney injury [fluid input_day1 ≥ 3390 ml (HR hazard ratio 1.42), fluid input_day2 ≥ 2734 ml (HR 1.64), platelet_min_day5 ≤ 224.2 × 109/l (HR 0.86), length of ICU stay ≥ 2.5 days (HR 1.24), length of hospital stay ≥ 5.8 days (HR 1.18), Bun_max_day1 ≥ 20 mmol/l (HR 1.20), and mechanical ventilation time ≥ 96 h (HR 1.11)] by multivariate Cox regression analysis, and the eleven predictors were entered into the nomogram. The nomogram model showed a discriminative ability for estimating sepsis-associated acute kidney injury. These results indicated that clinical parameters such as excess input fluid on the first and second days after admission and longer mechanical ventilation time could increase the risk of developing sepsis-associated acute kidney injury. With our study, we built a real-time prediction model for potentially forecasting acute kidney injury in septic patients that can help clinicians make decisions as early as possible to avoid sepsis-associated acute kidney injury.
Assuntos
Injúria Renal Aguda , Sepse , Adulto , Humanos , Estado Terminal , Nomogramas , Injúria Renal Aguda/diagnóstico , Injúria Renal Aguda/etiologia , Sepse/complicações , PlaquetasRESUMO
Mycotoxins are toxic secondary metabolites produced by filamentous fungi belonging, in particular, to the Aspergillus, Fusarium, and Penicillium genera [...].