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1.
Aquat Toxicol ; 218: 105359, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31765944

RESUMO

Carbohydrate metabolism switches from aerobic to anaerobic (glycolysis) to supply energy in response to acute hypoxic stress. Acute hypoxic stress with dissolved oxygen (DO) levels of 1.2 ±â€¯0.1 mg/L for 24 h and 12 h re-oxygenation was used to investigate the response of the anaerobic glycolytic pathway in Micropterus salmoides muscle. The results showed that the glucose concentration was significantly lower in muscle, while the lactic acid and pyruvic acid concentrations tended to increase during hypoxic stress. No significant difference was observed in muscle glycogen, and ATP content fluctuated significantly. The activities of gluconeogenesis-related enzymes were slightly elevated, such as phosphoenolpyruvate carboxykinase (PEPCK). The activities of the glycolytic enzymes increased after the induction of hypoxia, such as hexokinase (HK), pyruvate kinase (PK), and lactate dehydrogenase (LDH). Curiously, phosphofructokinase (PFK) activity was significantly down-regulated within 4 h during hypoxia, although these effects were transient, and most indices returned to control levels after 12 h of re-oxygenation. Upregulated hif-1α, ampkα, hk, glut1, and ldh mRNA expression suggested that carbohydrate metabolism was reprogrammed under hypoxia. Lactate transport was regulated by miR-124-5p according to quantitative polymerase chain reaction and dual luciferase reporter assays. Our findings provide new insight into the molecular regulatory mechanism of hypoxia in Micropterus salmoides muscle.


Assuntos
Aclimatação/fisiologia , Bass/metabolismo , Hipóxia/metabolismo , Ácido Láctico/metabolismo , MicroRNAs/genética , Transportadores de Ácidos Monocarboxílicos/genética , Músculos/metabolismo , Simportadores/genética , Aclimatação/genética , Animais , Bass/genética , Metabolismo dos Carboidratos/genética , Metabolismo dos Carboidratos/fisiologia , Regulação da Expressão Gênica , Hipóxia/genética , Músculos/enzimologia , Oxigênio/metabolismo
2.
Genes (Basel) ; 10(10)2019 10 12.
Artigo em Inglês | MEDLINE | ID: mdl-31614866

RESUMO

Gnetum possesses morphologically bisexual but functionally unisexual reproductive structures that exude sugary pollination drops to attract insects. Previous studies have revealed that the arborescent species (G. gnemon L.) and the lianoid species (G. luofuense C.Y.Cheng) possess different pollination syndromes. This study compared the proteome in the pollination drops of these two species using label-free quantitative techniques. The transcriptomes of fertile reproductive units (FRUs) and sterile reproductive units (SRUs) for each species were furthermore compared using Illumina Hiseq sequencing, and integrated proteomic and transcriptomic analyses were subsequently performed. Our results show that the differentially expressed proteins between FRUs and SRUs were involved in carbohydrate metabolism, the biosynthesis of amino acids and ovule defense. In addition, the differentially expressed genes between the FRUs and SRUs (e.g., MADS-box genes) were engaged in reproductive development and the formation of pollination drops. The integrated protein-transcript analyses revealed that FRUs and their exudates were relatively conservative while the SRUs and their exudates were more diverse, probably functioning as pollinator attractants. The evolution of reproductive organs appears to be synchronized with changes in the pollination drop proteome of Gnetum, suggesting that insect-pollinated adaptations are not restricted to angiosperms but also occur in gymnosperms.


Assuntos
Gnetum/metabolismo , Polinização/genética , Proteoma/metabolismo , Transcriptoma/genética , Aminoácidos/biossíntese , Aminoácidos/genética , Animais , Metabolismo dos Carboidratos/genética , Perfilação da Expressão Gênica , Gnetum/genética , Sequenciamento de Nucleotídeos em Larga Escala , Insetos/genética , Insetos/metabolismo , Proteínas de Domínio MADS/genética , Proteínas de Domínio MADS/metabolismo , Magnoliopsida/genética , Óvulo Vegetal/genética , Óvulo Vegetal/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Polinização/fisiologia , Proteoma/genética , Proteômica , Reprodução/genética , Açúcares/metabolismo , Transcriptoma/fisiologia
3.
PLoS Genet ; 15(10): e1008410, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31584940

RESUMO

Mitochondria have been increasingly recognized as a central regulatory nexus for multiple metabolic pathways, in addition to ATP production via oxidative phosphorylation (OXPHOS). Here we show that inducing mitochondrial DNA (mtDNA) stress in Drosophila using a mitochondrially-targeted Type I restriction endonuclease (mtEcoBI) results in unexpected metabolic reprogramming in adult flies, distinct from effects on OXPHOS. Carbohydrate utilization was repressed, with catabolism shifted towards lipid oxidation, accompanied by elevated serine synthesis. Cleavage and translocation, the two modes of mtEcoBI action, repressed carbohydrate rmetabolism via two different mechanisms. DNA cleavage activity induced a type II diabetes-like phenotype involving deactivation of Akt kinase and inhibition of pyruvate dehydrogenase, whilst translocation decreased post-translational protein acetylation by cytonuclear depletion of acetyl-CoA (AcCoA). The associated decrease in the concentrations of ketogenic amino acids also produced downstream effects on physiology and behavior, attributable to decreased neurotransmitter levels. We thus provide evidence for novel signaling pathways connecting mtDNA to metabolism, distinct from its role in supporting OXPHOS.


Assuntos
Reprogramação Celular/genética , DNA Mitocondrial/genética , Diabetes Mellitus Tipo 2/genética , Mitocôndrias/genética , Trifosfato de Adenosina/genética , Animais , Metabolismo dos Carboidratos/genética , Carboidratos/genética , Enzimas de Restrição do DNA/genética , Diabetes Mellitus Tipo 2/metabolismo , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Humanos , Redes e Vias Metabólicas/genética , Mitocôndrias/metabolismo , Fosforilação Oxidativa , Estresse Oxidativo/genética
4.
Microb Cell Fact ; 18(1): 134, 2019 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-31409414

RESUMO

BACKGROUND: The halophilic bacterium Chromohalobacter salexigens metabolizes glucose exclusively through the Entner-Doudoroff (ED) pathway, an adaptation which results in inefficient growth, with significant carbon overflow, especially at low salinity. Preliminary analysis of C. salexigens genome suggests that fructose metabolism could proceed through the Entner-Doudoroff and Embden-Meyerhof-Parnas (EMP) pathways. In order to thrive at high salinity, this bacterium relies on the biosynthesis and accumulation of ectoines as major compatible solutes. This metabolic pathway imposes a high metabolic burden due to the consumption of a relevant proportion of cellular resources, including both energy molecules (NADPH and ATP) and carbon building blocks. Therefore, the existence of more than one glycolytic pathway with different stoichiometries may be an advantage for C. salexigens. The aim of this work is to experimentally characterize the metabolism of fructose in C. salexigens. RESULTS: Fructose metabolism was analyzed using in silico genome analysis, RT-PCR, isotopic labeling, and genetic approaches. During growth on fructose as the sole carbon source, carbon overflow was not observed in a wide range of salt concentrations, and higher biomass yields were reached. We unveiled the initial steps of the two pathways for fructose incorporation and their links to central metabolism. While glucose is metabolized exclusively through the Entner-Doudoroff (ED) pathway, fructose is also partially metabolized by the Embden-Meyerhof-Parnas (EMP) route. Tracking isotopic label from [1-13C] fructose to ectoines revealed that 81% and 19% of the fructose were metabolized through ED and EMP-like routes, respectively. Activities of enzymes from both routes were demonstrated in vitro by 31P-NMR. Genes encoding predicted fructokinase and 1-phosphofructokinase were cloned and the activities of their protein products were confirmed. Importantly, the protein encoded by csal1534 gene functions as fructose bisphosphatase, although it had been annotated previously as pyrophosphate-dependent phosphofructokinase. The gluconeogenic rather than glycolytic role of this enzyme in vivo is in agreement with the lack of 6-phosphofructokinase activity previously described. CONCLUSIONS: Overall, this study shows that C. salexigens possesses a greater metabolic flexibility for fructose catabolism, the ED and EMP pathways contributing to a fine balancing of energy and biosynthetic demands and, subsequently, to a more efficient metabolism.


Assuntos
Chromohalobacter/genética , Chromohalobacter/metabolismo , Frutose/metabolismo , Glicólise , Metabolismo dos Carboidratos/genética , Carbono/metabolismo , Genoma Bacteriano , Glucose/metabolismo , Redes e Vias Metabólicas , Salinidade
5.
Int J Mol Med ; 44(4): 1495-1504, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31432104

RESUMO

Post­transplantation diabetes mellitus (PTDM) is a known side effect in transplant recipients administered with immunosuppressant drugs, such as tacrolimus (Tac). Although injury of islet cells is considered a major reason for Tac­induced PTDM, the involvement of insulin resistance in PTDM remains unknown. In the present study, expression levels of adipocytokines, glucose metabolism associated genes and peroxisome proliferator­activated receptor (PPAR)­Î³ in adipose, muscular and liver tissues from a rat model induced with Tac (1 mg/kg/day) were examined. Rats developed hyperglycemia and glucose intolerance after 10 days of Tac administration. A subgroup of diabetic rats was further treated with rosiglitazone (4 mg/kg), a PPAR­Î³ activator. Adipose, muscle and liver tissues were obtained on day 15 after induction and the results demonstrated that expression levels of adipocytokines, PPAR­Î³ and proteins in the insulin associated signaling pathway varied in the different groups. Rosiglitazone administration significantly improved hyperglycemia, glucose intolerance and expression levels of proteins associated with insulin signaling, as well as adipocytokines expression. The results of this study demonstrated that adipocytokines and PPAR­Î³ signaling may serve important roles in the pathogenesis of Tac­induced PTDM, which may provide a promising application in the treatment of PTDM in the future.


Assuntos
Metabolismo dos Carboidratos/genética , Diabetes Mellitus Experimental/etiologia , Diabetes Mellitus Experimental/metabolismo , Regulação da Expressão Gênica , Glucose/metabolismo , Transplante de Órgãos/efeitos adversos , Tacrolimo/efeitos adversos , Animais , Biomarcadores , Modelos Animais de Doenças , Expressão Gênica , Intolerância à Glucose , Imunossupressores/efeitos adversos , Insulina/metabolismo , Masculino , Especificidade de Órgãos , Ratos , Transdução de Sinais
6.
BMC Genomics ; 20(1): 657, 2019 Aug 17.
Artigo em Inglês | MEDLINE | ID: mdl-31419936

RESUMO

BACKGROUND: Garlic bulbs are abnormal expanding axillary buds that are rarely found among vascular plants. Bulb-yield is one of the valuable agronomic traits of garlic. However, due to the large genome size and a strictly asexual life cycle in the cultivars, the genetic basis of the yield traits are poorly understood in garlic. RESULTS: In the present study, we carried out an association mapping for three yield traits of garlic bulbs: bulb weight (BW), diameter (BD), and the number of garlic cloves (CN), using the recently proposed transcriptome-referenced association study. In total 25, 2, and 30 single nucleotide polymorphisms (SNPs), were identified in the transcripts to be associated with BW, BD, and CN traits, respectively. Of the transcripts with associated SNPs, the expression of 17 of them showed a significant correlation with the corresponding traits in the population, suggesting their relation to bulbs yield traits. Six transcripts were long non-coding RNAs (lncRNAs), and the others encode proteins involved mainly in carbohydrate metabolism, transcription regulation, cytokinin activity, protein degradation, etc. In addition, expression quantitative trait locus (eQTL) and expression correlation analysis have revealed that seven CN-related transcripts displayed interrelation, constituting two potential pathways. CONCLUSION: This study provides novel insights into the genetic basis of the yield traits in garlic bulbs, and the identification of trait-associated SNPs/transcripts provides a basis for improving the bulb yield in garlic breeding.


Assuntos
Alho/genética , Raízes de Plantas/genética , Metabolismo dos Carboidratos/genética , Citocininas/genética , Citocininas/metabolismo , Alho/crescimento & desenvolvimento , Alho/metabolismo , Regulação da Expressão Gênica de Plantas , Fenótipo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Polimorfismo de Nucleotídeo Único , Proteólise , Locos de Características Quantitativas/genética , RNA Longo não Codificante/metabolismo , Transcriptoma
7.
J Diabetes Res ; 2019: 5451038, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31467927

RESUMO

Aims: Posttranslational modifications of histones and transcription factors regulate gene expression and are implicated in beta-cell failure and diabetes. We have recently shown that preserving H3K27 and H3K4 methylation using the lysine demethylase inhibitor GSK-J4 reduces cytokine-induced destruction of beta-cells and improves beta-cell function. Here, we investigate the therapeutic potential of GSK-J4 to prevent diabetes development and examine the importance of H3K4 methylation for islet function. Materials and Methods: We used two mouse models of diabetes to investigate the therapeutic potential of GSK-J4. To clarify the importance of H3K4 methylation, we characterized a mouse strain with knockout (KO) of the H3K4 demethylase KDM5B. Results: GSK-J4 administration failed to prevent the development of experimental diabetes induced by multiple low-dose streptozotocin or adoptive transfer of splenocytes from acutely diabetic NOD to NODscid mice. KDM5B-KO mice were growth retarded with altered body composition, had low IGF-1 levels, and exhibited reduced insulin secretion. Interestingly, despite secreting less insulin, KDM5B-KO mice were able to maintain normoglycemia following oral glucose tolerance test, likely via improved insulin sensitivity, as suggested by insulin tolerance testing and phosphorylation of proteins belonging to the insulin signaling pathway. When challenged with high-fat diet, KDM5B-deficient mice displayed similar weight gain and insulin sensitivity as wild-type mice. Conclusion: Our results show a novel role of KDM5B in metabolism, as KDM5B-KO mice display growth retardation and improved insulin sensitivity.


Assuntos
Metabolismo dos Carboidratos/genética , Proteínas de Ligação a DNA/fisiologia , Glucose/metabolismo , Células Secretoras de Insulina/fisiologia , Ilhotas Pancreáticas/fisiologia , Histona Desmetilases com o Domínio Jumonji/fisiologia , Animais , Proteínas de Ligação a DNA/genética , Diabetes Mellitus Experimental/induzido quimicamente , Diabetes Mellitus Experimental/genética , Diabetes Mellitus Experimental/metabolismo , Transtornos do Crescimento/genética , Transtornos do Crescimento/metabolismo , Homeostase/genética , Resistência à Insulina/genética , Células Secretoras de Insulina/metabolismo , Ilhotas Pancreáticas/metabolismo , Histona Desmetilases com o Domínio Jumonji/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Camundongos Knockout , Camundongos SCID , Estreptozocina
8.
Int J Mol Sci ; 20(15)2019 Aug 04.
Artigo em Inglês | MEDLINE | ID: mdl-31382684

RESUMO

About 15% of higher plants have acquired the ability to convert sucrose into fructans. Fructan degradation is catalyzed by fructan exohydrolases (FEHs), which are structurally related to cell wall invertases (CWI). However, the biological function(s) of FEH enzymes in non-fructan species have remained largely enigmatic. In the present study, one maize CWI-related enzyme named Zm-6&1-FEH1, displaying FEH activity, was explored with respect to its substrate specificities, its expression during plant development, and its possible interaction with CWI inhibitor protein. Following heterologous expression in Pichia pastoris and in N. benthamiana leaves, recombinant Zm-6&1-FEH1 revealed substrate specificities of levan and inulin, and also displayed partially invertase activity. Expression of Zm-6&1-FEH1 as monitored by qPCR was strongly dependent on plant development and was further modulated by abiotic stress. To explore whether maize FEH can interact with invertase inhibitor protein, Zm-6&1-FEH1 and maize invertase inhibitor Zm-INVINH1 were co-expressed in N. benthamiana leaves. Bimolecular fluorescence complementation (BiFC) analysis and in vitro enzyme inhibition assays indicated productive complex formation. In summary, the results provide support to the hypothesis that in non-fructan species FEH enzymes may modulate the regulation of CWIs.


Assuntos
Glicosídeo Hidrolases/genética , Folhas de Planta/enzimologia , Zea mays/enzimologia , beta-Frutofuranosidase/genética , Sequência de Aminoácidos , Metabolismo dos Carboidratos/genética , Frutanos/genética , Frutanos/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Glicosídeo Hidrolases/química , Folhas de Planta/genética , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Especificidade por Substrato , Tabaco/enzimologia , Tabaco/genética , Zea mays/genética , beta-Frutofuranosidase/antagonistas & inibidores
9.
Insect Biochem Mol Biol ; 112: 103184, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31295549

RESUMO

The rate of carbohydrate metabolism is tightly coordinated with developmental transitions in Drosophila, and fluctuates depending on the requirements of a particular developmental stage. These successive metabolic switches result from changes in the expression levels of genes encoding glycolytic, tricarboxylic acid cycle (TCA), and oxidative phosphorylation enzymes. In this report, we describe a repressive action of ecdysone signaling on the expression of glycolytic genes and enzymes of glycogen metabolism in Drosophila development. The basis of this effect is an interaction between the ecdysone receptor (EcR) and the estrogen-related receptor (ERR), a specific regulator of the Drosophila glycolysis. We found an overlapping DNA-binding pattern for the EcR and ERR in the Drosophila S2 cells. EcR was detected at a subset of the ERR target genes responsible for carbohydrate metabolism. The 20-hydroxyecdysone treatment of both the Drosophila larvae and the S2 cells decreased transcriptional levels of ERR targets. We propose a joint action mode for both the EcR and ERR, for at least a subset of the glycolytic genes. We find that both receptors bind to the same regulatory regions and may form or be part of a joint transcriptional regulatory complex in the Drosophila S2 cells.


Assuntos
Metabolismo dos Carboidratos/genética , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Receptores Estrogênicos/metabolismo , Receptores de Esteroides/metabolismo , Animais , Linhagem Celular , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/crescimento & desenvolvimento , Ecdisterona/farmacologia , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Larva/crescimento & desenvolvimento , Larva/metabolismo , Receptores Citoplasmáticos e Nucleares/metabolismo , Receptores Estrogênicos/genética , Receptores de Esteroides/genética
10.
Oncogene ; 38(34): 6211-6225, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31289360

RESUMO

One-carbon metabolism plays a central role in a broad array of metabolic processes required for the survival and growth of tumor cells. However, the molecular basis of how one-carbon metabolism may influence RNA methylation and tumorigenesis remains largely unknown. Here we show MTHFD2, a mitochondrial enzyme involved in one-carbon metabolism, contributes to the progression of renal cell carcinoma (RCC) via a novel epitranscriptomic mechanism that involves HIF-2α. We found that expression of MTHFD2 was significantly elevated in human RCC tissues, and MTHFD2 knockdown strongly reduced xenograft tumor growth. Mechanistically, using an unbiased methylated RNA immunoprecipitation sequencing (meRIP-Seq) approach, we found that MTHFD2 plays a critical role in controlling global N6-methyladenosine (m6A) methylation levels, including the m6A methylation of HIF-2α mRNA, which results in enhanced translation of HIF-2α. Enhanced HIF-2α translation, in turn, promotes the aerobic glycolysis, linking one-carbon metabolism to HIF-2α-dependent metabolic reprogramming through RNA methylation. Our findings also suggest that MTHFD2 and HIF-2α form a positive feedforward loop in RCC, promoting metabolic reprograming and tumor growth. Taken together, our results suggest that MTHFD2 links RNA methylation status to the metabolic state of tumor cells in RCC.


Assuntos
Aminoidrolases/fisiologia , Carcinoma de Células Renais/metabolismo , Glicólise/genética , Neoplasias Renais/metabolismo , Metilenotetra-Hidrofolato Desidrogenase (NADP)/fisiologia , Metiltransferases/metabolismo , Enzimas Multifuncionais/fisiologia , Processamento Pós-Transcricional do RNA/genética , Animais , Metabolismo dos Carboidratos/genética , Carcinoma de Células Renais/genética , Carcinoma de Células Renais/patologia , Linhagem Celular Tumoral , Reprogramação Celular/genética , Regulação Neoplásica da Expressão Gênica , Humanos , Neoplasias Renais/genética , Neoplasias Renais/patologia , Masculino , Metilação , Camundongos , Camundongos Nus
11.
Biomed Res Int ; 2019: 1493721, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31321228

RESUMO

The mechanism of autolysis of Volvariella volvacea (V. volvacea) at low temperature has not been fully explained. As mannitol is among the most important osmotic adjustment substances in fungal resistance, this study sampled mycelia of strains V23 and VH3 treated at 0°C for 0, 2, 4, 8, and 10 h to analyze changes in intracellular mannitol content by high-performance anion chromatography with pulsed amperometric detection (HAPEC-PAD). Reverse transcription quantitative PCR (RT-qPCR) analysis was applied to assess differences in the transcript levels of genes associated with mannitol metabolism under low-temperature stress. A mannitol solution was added to cultures of V. volvacea fruiting bodies, and effects on the hypothermic resistance of these organs were explored by evaluating variations in sensory properties during cryogenic storage after harvest. The results suggested that in the initial stage of low-temperature treatment, intracellular mannitol was largely catabolized as an energy storage material and the expression of genes encoding enzymes involved in synthetic reactions was inhibited. However, low-temperature resistance was induced with further treatment, with activation of mannitol synthesis and inhibition of degradation; the cells accumulated mannitol, leading to osmoregulation. No significant elongation of V. volvacea fruiting bodies during storage at 4°C was observed, and these organs tended to shrink and collapse. The sensory quality of mannitol-treated fruiting bodies was much better than that of control fruiting bodies. Application of a mannitol solution at the cultivation stage of V. volvacea somewhat improved the low-temperature resistance of the fruiting bodies, verifying the correlation between mannitol and resistance to this stress in V. volvacea. The results of this study lay a foundation for a deeper understanding of the autolysis mechanism of V. volvacea, providing technical support for increasing the cryopreservation time of this species and extending the postharvest shelf life of its fruiting bodies. In addition, the mechanism underlying the low-temperature tolerance of the VH3 strain should be further explained at the molecular level.


Assuntos
Proteínas Fúngicas/genética , Manitol/metabolismo , Filogenia , Volvariella/metabolismo , Metabolismo dos Carboidratos/genética , Temperatura Baixa , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Volvariella/genética
12.
Biomed Pharmacother ; 117: 108989, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31228801

RESUMO

Korean pine nut protein (PNP) has a variety of biological activities, which are good for human health, but its ability to preventing diabetes has not been reported. This study evaluated the effects of water-soluble proteins of Korean pine nut obtained from a dilute alkali extract on carbohydrate metabolism of type 2 diabetic mice on a model of diabetes induced using a high fat diet combined with streptozotocin. The results showed that the hypoglycemic effect of PNP at a middle dose was the most significant, which was 38.7% lower than that of control. The extract significantly improved the oral glucose tolerance and liver indexes, increased the activity of the carbohydrate metabolism enzymes, and regulated the expression of the function of key genes for carbohydrate metabolism. It had a positive effect on both insulin resistance and glycolytic/gluconeogenesis signaling. In conclusion, PNP can regulate fasting blood glucose, improve insulin resistance, correct the glucose metabolism disorder in diabetic mice, and have a positive regulatory role. As the functional food, it has the potential to be beneficial in the treatment of type 2 diabetes mellitus as a new hypoglycemic functional food.


Assuntos
Diabetes Mellitus Tipo 2/tratamento farmacológico , Hipoglicemiantes/uso terapêutico , Proteínas de Nozes/uso terapêutico , Pinus/química , Água/química , Administração Oral , Aminoácidos/análise , Animais , Glicemia/metabolismo , Peso Corporal/efeitos dos fármacos , Metabolismo dos Carboidratos/efeitos dos fármacos , Metabolismo dos Carboidratos/genética , Diabetes Mellitus Tipo 2/sangue , Dieta , Comportamento de Ingestão de Líquido , Jejum/sangue , Comportamento Alimentar , Regulação da Expressão Gênica/efeitos dos fármacos , Teste de Tolerância a Glucose , Glicogênio/metabolismo , Hipoglicemiantes/administração & dosagem , Hipoglicemiantes/farmacologia , Insulina/sangue , Resistência à Insulina , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fígado/patologia , Masculino , Camundongos , Músculos/efeitos dos fármacos , Músculos/metabolismo , Proteínas de Nozes/administração & dosagem , Proteínas de Nozes/farmacologia , Solubilidade
13.
Vopr Pitan ; 88(2): 6-16, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31233683

RESUMO

Quercetin (Q; 3,3',4',5,7-pentahydroxyflavone) is considered as a promising component of specialized products for the correction of metabolic disorders in obesity and metabolic syndrome. At the same time, the results of evaluating the clinical efficacy of Q are ambiguous, and the mechanisms of its influence on lipid and carbohydrate-energy metabolism are not well understood. The aim of the work was to study the effect of quercetin (Q 3,3',4',5,7-pentahydroxyflavone) on the expression of key glycolysis and lipogenesis enzymes' genes in Zucker-Leprfa (Z) rats characterized by hereditary obesity, compared to «wild-type¼ Wistar (W) rats. Material and methods. 24 male Z rats and 32 male W rats aged 8-10 weeks were used. Animals of each line were divided into 4 groups of equal numbers. For 62 days the animals of the first groups (controls) received a balanced diet according to AIN93M, the seconds - the same diet with Q added in a dose of 50 mg/kg body weight. Animals of the third groups received a high-fat, high-carbohydrate diet (HFCD) with fat 30% by weight and with the replacement of drinking water with a 20% solution of fructose, the fourths groups - the same diet and supplementation with Q. After removing animals from the experiment, expression levels of liver carbohydrate and lipid metabolism genes Khk, Gck, Pklr, Acaca, Fasn, Scd, Srebf1, Mlxipl, Ppara and Pparg were determined by real-time polymerase chain reaction (RT-PCR) with reverse transcription using Actb and Gapdh as reference genes. The levels of triglycerides, total and HDL cholesterol, lipolytic activity and immunoreactive leptin were determined in plasma. Results and discussion. When comparing two animal lines, a significantly higher level of expression of Ppara, Pparg, Mlxipl, Acaca, Fasn, Scd was shown in Z rats compared to W rats, which is consistent with the development of dyslipidemia in the first ones and elevated levels of leptin under both types of diets used. The addition of Q caused in W rats a decrease in the expression of Scd, Mlxipl, Khk and Gck, more pronounced on the background of HFCD whereas in Z rats there were no similar effects, or they had the opposite direction. In addition, in Z rats, consumption of Q led to increased expression of Pklr, which was not observed in W rats. Conclusion. The modulating effect of Q on the expression of key genes of lipid and carbohydrate metabolism enzymes significantly differs in wild-type W rats and mutant Z rats with hereditary obesity, and this difference appears to be potentiated by the consumption of excess fat and fructose.


Assuntos
Metabolismo dos Carboidratos/efeitos dos fármacos , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/metabolismo , Quercetina/farmacologia , Animais , Metabolismo dos Carboidratos/genética , Metabolismo dos Lipídeos/genética , Masculino , Ratos , Ratos Wistar , Ratos Zucker , Especificidade da Espécie
14.
BMC Genomics ; 20(1): 480, 2019 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-31185888

RESUMO

BACKGROUND: Plasmids play an vital role in driving the rapid global spread of antimicrobial resistance and adaptation to changing ambient conditions. It has been suggested that the presence of plasmids can pose tremendous impacts on the host physiology. However, little is known regarding the contributions of carbapenemase-encoding plasmid carriage on the physiology and pathogenicity of hypervirulent K. pneumoniae (hvKP). RESULTS: Here we performed a transcriptomic analysis of hvKP with or without carbapenemase-encoding plasmid p24835-NDM5. The results had shown 683 genes with differential expression (false discovery rate, ≤0.001; > 2-fold change), of which 107 were up-regulated and 576 were down-regulated. Gene groups with functions relating to carbohydrate metabolism and multidrug efflux system were increased in genes with increased expression, and those relating to capsule biosynthesis and virulence factors were increased in the genes with decreased expression. In agreement with these changes, survival rate of TfpNDM-hvKP in the presence of normal human serum decreased, and competitive index (CI values) indicated significant fitness defects in the plasmid-carrying hvKP strain when co-cultured with its plasmid-free isogenic ancestor and the ATCC control. Moreover, the p24835-NDM5-containing hvKP strain retained its high neutrophil-mediated phagocytosis and murine lethality. CONCLUSION: These data indicate that hvKP responds to carbapenemase-encoding plasmid by altering the expression of genes involved in carbohydrate metabolism, antibiotic resistance, capsule biosynthesis and virulence expression. Apart from antibiotic resistance selective advantages, carbapenemase-encoding plasmid carriage may also lead to virulence change or adaption to specific habitats in hvKP strain.


Assuntos
Proteínas de Bactérias/genética , Perfilação da Expressão Gênica , Klebsiella pneumoniae/genética , Fenótipo , Plasmídeos/genética , beta-Lactamases/genética , Adulto , Animais , Antibacterianos/farmacologia , Biofilmes/crescimento & desenvolvimento , Metabolismo dos Carboidratos/genética , Humanos , Cinética , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/metabolismo , Klebsiella pneumoniae/patogenicidade , Camundongos , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Fagocitose , Virulência
15.
Plant Physiol Biochem ; 141: 164-171, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31170640

RESUMO

For a comprehensive understanding of gene expression, enzyme activity and sugar concentrations in response to short-term water deficit in apple (Greensleeves), sugar-modulated gene expression and enzyme activities were analyzed. Water stress resulted in the accumulation of sorbitol, glucose, fructose, galactose and starch, accompanied by a significant reduction in photosynthesis and sucrose concentration. In response to short-term water deficits, the activities of aldose-6-phosphate reductase (A6PR; EC 1.1.1.200), sorbitol dehydrogenase (SDH; EC 1.1.1.14), neutral invertase (NINV; EC 3.2.1.26), sucrose synthase (SUSY; EC 2.4.1.13), and fructokinase (FK; EC 2.7.1.4) were higher, whereas cell wall invertase (CWINV; EC 3.2.1.26) and hexokinase (HK; EC 2.7.1.1) activities were lower. In addition, sucrose phosphate synthase (SPS; EC 2.4.1.14) activity increased during the initial stages of dehydration and then decreased as the drought strengthened. Transcript levels of MdA6PR, MdSDH1/2, MdNINV1/2, MdSUSY3, MdFK1/2/4, MdSOT1/2, MdSUC1-3, MdTMT2/3, MdvGT1, MdpGlcT1-4 were upregulated, whereas transcript levels of MdCWINV1/2, MdHK1/2/3/5, and MdTMT1 were downregulated after 6 days of water stress. These findings suggest that the sorbitol metabolism pathway is induced and high levels of hexose derived from photosynthetic products are transported into vacuoles for adjustment to the water deficit. Our results provide insights into the relationships between sugar levels and sugar-modulated gene and enzyme activity in response to the imposition of short-term water stress.


Assuntos
Metabolismo dos Carboidratos/genética , Secas , Regulação da Expressão Gênica de Plantas , Malus/fisiologia , Folhas de Planta/fisiologia , Parede Celular/metabolismo , Frutas/metabolismo , Glucosiltransferases/metabolismo , Malus/genética , Fotossíntese , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Sorbitol/metabolismo , Estresse Fisiológico , Sacarose/metabolismo , beta-Frutofuranosidase/metabolismo
16.
Bull Exp Biol Med ; 167(2): 263-266, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31243677

RESUMO

We studied the expression of genes encoding enzymes of carbohydrate and lipid metabolism ketohexokinase (Khk), glucokinase (Gck), pyruvate kinase (Pklr), acetyl-Co-carboxylase (Acaca), fatty acid synthase (Fasn), stearoyl-CoA desaturase (Scd), and their transcription regulators ChREBP (Mlxipl), SREBP-1c (Srebf1), and PPARα (Ppara) in rat liver. Control group rats received a semisynthetic ration over 20 weeks. Experimental group 1 received a semisynthetic ration and 20% fructose solution instead of drinking water. Experimental group 2 rats received a semisynthetic ration with quercetin (0.1% fodder weight) and 20% fructose solution. Consumption of 20% fructose solution (experimental group 1) led to an increase in Scd expression in comparison with the control and did not affect the expression of other genes. Addition of quercetin to the ration (experimental group 2) led to a decrease in the expression of Khk, Gck, Fasn, Scd, Mlxipl, and Ppara genes in comparison with experimental group 1. The results suggest that quercetin reduced the expression of genes of carbohydrate and lipid metabolism enzymes in the liver of rats receiving high-fructose ration.


Assuntos
Enzimas/genética , Frutose/metabolismo , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/enzimologia , Quercetina/farmacologia , Animais , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/metabolismo , Metabolismo dos Carboidratos/efeitos dos fármacos , Metabolismo dos Carboidratos/genética , Dieta , Enzimas/metabolismo , Frutoquinases/genética , Frutoquinases/metabolismo , Glucoquinase/genética , Glucoquinase/metabolismo , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Masculino , Piruvato Quinase/genética , Piruvato Quinase/metabolismo , Ratos , Ratos Wistar , Estearoil-CoA Dessaturase/genética , Estearoil-CoA Dessaturase/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo
17.
EMBO J ; 38(12)2019 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-31101674

RESUMO

Transcription factors show rapid and reversible binding to chromatin in living cells, and transcription occurs in sporadic bursts, but how these phenomena are related is unknown. Using a combination of in vitro and in vivo single-molecule imaging approaches, we directly correlated binding of the Gal4 transcription factor with the transcriptional bursting kinetics of the Gal4 target genes GAL3 and GAL10 in living yeast cells. We find that Gal4 dwell time sets the transcriptional burst size. Gal4 dwell time depends on the affinity of the binding site and is reduced by orders of magnitude by nucleosomes. Using a novel imaging platform called orbital tracking, we simultaneously tracked transcription factor binding and transcription at one locus, revealing the timing and correlation between Gal4 binding and transcription. Collectively, our data support a model in which multiple RNA polymerases initiate transcription during one burst as long as the transcription factor is bound to DNA, and bursts terminate upon transcription factor dissociation.


Assuntos
Nucleossomos/metabolismo , Fatores de Transcrição/metabolismo , Ativação Transcricional , Sítios de Ligação , Metabolismo dos Carboidratos/genética , Galactoquinase/genética , Galactoquinase/metabolismo , Galactose/metabolismo , Regulação Fúngica da Expressão Gênica , Imagem Molecular/métodos , Organismos Geneticamente Modificados , Ligação Proteica , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Análise de Célula Única/métodos , Transativadores/genética , Transativadores/metabolismo , Fatores de Transcrição/genética , Transcrição Genética , Ativação Transcricional/genética
18.
J Microbiol ; 57(7): 575-586, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31073898

RESUMO

Mangrove sediment microorganisms play a vital role in the energy transformation and element cycling in marine wetland ecosystems. Using metagenomics analysis strategy, we compared the taxonomic structure and gene profile of the mangrove and non-mangrove sediment samples at the subtropical estuary in Beibu Gulf, South China Sea. Proteobacteria, Bacteroidetes, and Firmicutes were the most abundant bacterial phyla. Archaeal family Methanosarcinaceae and bacterial genera Vibrio and Dehalococcoides were significantly higher in the mangrove sediments than in the nonmangrove sediments. Functional analysis showed that "Carbohydrate metabolism" was the most abundant metabolic category. The feature of carbohydrate-active enzymes (CZs) was analyzed using the Carbohydrate-Active EnZymes Database. The significant differences of CZs between mangrove and non-mangrove sediments, were attributed to the amounts of polyphenol oxidase (EC 1.10.3.-), hexosyltransferase (EC 2.4.1.-), and ß-N-acetylhexosaminidase (EC 3.2.1.52), which were higher in the mangrove sediment samples. Principal component analysis indicated that the microbial community and gene profile between mangrove and non-mangrove sediments were distinct. Redundancy analysis showed that total organic carbon is a significant factor that affects the microbial community and gene distribution. The results indicated that the mangrove ecosystem with massive amounts of organic carbon may promote the richness of carbohydrate metabolism genes and enhance the degradation and utilization of carbohydrates in the mangrove sediments.


Assuntos
Archaea , Bactérias , Sedimentos Geológicos/microbiologia , Archaea/classificação , Archaea/genética , Archaea/metabolismo , Bactérias/classificação , Bactérias/genética , Bactérias/metabolismo , Metabolismo dos Carboidratos/genética , China , Metagenômica/métodos , Microbiota , Filogenia , RNA Ribossômico 16S/genética , Áreas Alagadas
19.
Int J Mol Sci ; 20(7)2019 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-30934901

RESUMO

Massively input and accumulated ammonium is one of the main causes of eutrophication in aquatic ecosystems, which severely deteriorates water quality. Previous studies showed that one of the commonly used macrophytes, Myriophyllum aquaticum, was capable of not only withstanding ammonium of high concentration, but also efficiently assimilating extracellular ammonium to constitutive amino acids and proteins. However, the genetic mechanism regulating such efficient nitrogen metabolism in M. aquaticum is still poorly understood. Therefore, RNA-based analysis was performed in this study to understand the ammonium regulatory mechanism in M. aquaticum in response to various concentrations of ammonium. A total of 7721 genes were differentially expressed, of which those related to nitrogen-transport, assimilation, and remobilization were highly-regulated in response to various concentrations of ammonium. We have also identified transcription factors and protein kinases that were rapidly induced in response to ammonium, which suggests their involvement in ammonium-mediated signalling. Meanwhile, secondary metabolism including phenolics and anthocyanins biosynthesis was also activated in response to various concentrations of ammonium, especially at high ammonium concentrations. These results proposed a complex physiological and genetic regulation network related to nitrogen, carbohydrate, transcription factors, and secondary metabolism for nitrogen use efficiency in M. aquaticum.


Assuntos
Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Genes de Plantas , Nitrogênio/metabolismo , Saxifragales/genética , Análise de Sequência de RNA , Transcriptoma/genética , Aminoácidos/análise , Compostos de Amônio/farmacologia , Metabolismo dos Carboidratos/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ontologia Genética , Família Multigênica , Fenótipo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Reprodutibilidade dos Testes , Fatores de Transcrição/metabolismo
20.
Mol Cell Endocrinol ; 490: 47-56, 2019 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-30974146

RESUMO

The liver helps maintain energy homeostasis by synthesizing and storing glucose and lipids. Gonadal steroids, particularly estrogens, play an important role in regulating metabolism. As estrogens are considered female hormones, metabolic disorders related to the disruption of estrogen signaling have mostly been studied in females. Estrogen receptor alpha (ESR1) is the predominant receptor in both the male and female liver, and it mediates the hepatic response to estrogens. Loss of ESR1 increases weight gain and obesity in female rats, while reducing the normal growth in males. Although Esr1-/- male rats have a reduced body weight, they exhibit increased adipose deposition and impaired glucose tolerance. We further investigated whether these metabolic disorders in Esr1-/- male rats were linked with the loss of transcriptional regulation by ESR1 in the liver. To identify the ESR-regulated genes, RNA-sequencing was performed on liver mRNAs from wildtype and Esr1-/- male rats. Based on an absolute fold change of ≥2 with a p-value ≤ 0.05, a total of 706 differentially expressed genes were identified in the Esr1-/- male liver: 478 downregulated, and 228 upregulated. Pathway analyses demonstrate that the differentially expressed genes include transcriptional regulators (Cry1, Nr1d1, Nr0b2), transporters (Slc1a2), and regulators of biosynthesis (Cyp7b1, Cyp8b1), and hormone metabolism (Hsd17b2, Sult1e1). Many of these genes are also integral parts of the lipid and carbohydrate metabolism pathways in the liver. Interestingly, certain critical regulators of the metabolic pathways displayed a sexual dimorphism in expression, which may explain the divergent weight gain in Esr1-/- male and female rats despite common metabolic dysfunctions.


Assuntos
Metabolismo dos Carboidratos/genética , Receptor alfa de Estrogênio/metabolismo , Regulação da Expressão Gênica , Metabolismo dos Lipídeos/genética , Fígado/metabolismo , Adiposidade , Animais , Feminino , Ontologia Genética , Glucose/metabolismo , Insulina/metabolismo , Lipídeos/sangue , Masculino , Modelos Biológicos , Ratos Sprague-Dawley , Reprodutibilidade dos Testes , Ganho de Peso
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