Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 4.309
Filtrar
1.
Int J Mol Sci ; 22(18)2021 Sep 14.
Artigo em Inglês | MEDLINE | ID: mdl-34576076

RESUMO

Mesenteric ischemia and reperfusion (I/R) injury can ensue from a variety of vascular diseases and represents a major cause of morbidity and mortality in intensive care units. It causes an inflammatory response associated with local gut dysfunction and remote organ injury. Adenosine monophosphate-activated protein kinase (AMPK) is a crucial regulator of metabolic homeostasis. The catalytic α1 subunit is highly expressed in the intestine and vascular system. In loss-of-function studies, we investigated the biological role of AMPKα1 in affecting the gastrointestinal barrier function. Male knock-out (KO) mice with a systemic deficiency of AMPKα1 and wild-type (WT) mice were subjected to a 30 min occlusion of the superior mesenteric artery. Four hours after reperfusion, AMPKα1 KO mice exhibited exaggerated histological gut injury and impairment of intestinal permeability associated with marked tissue lipid peroxidation and a lower apical expression of the junction proteins occludin and E-cadherin when compared to WT mice. Lung injury with neutrophil sequestration was higher in AMPKα1 KO mice than WT mice and paralleled with higher plasma levels of syndecan-1, a biomarker of endothelial injury. Thus, the data demonstrate that AMPKα1 is an important requisite for epithelial and endothelial integrity and has a protective role in remote organ injury after acute ischemic events.


Assuntos
Proteínas Quinases Ativadas por AMP/deficiência , Lesão Pulmonar Aguda/complicações , Intestinos/enzimologia , Intestinos/lesões , Isquemia Mesentérica/complicações , Traumatismo por Reperfusão/complicações , Proteínas Quinases Ativadas por AMP/genética , Lesão Pulmonar Aguda/enzimologia , Animais , Caderinas/metabolismo , Permeabilidade da Membrana Celular , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Células Epiteliais/metabolismo , Glicocálix/metabolismo , Intestinos/patologia , Isquemia Mesentérica/enzimologia , Camundongos Endogâmicos C57BL , Ocludina/metabolismo , Traumatismo por Reperfusão/enzimologia
2.
Fish Physiol Biochem ; 47(4): 1243-1255, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34226986

RESUMO

The effects of stocking density on growth performance, serum biochemistry, digestive enzymes, immune response, and muscle quality of largemouth bass (Micropterus salmoides) reared in nine in-pond raceway systems (IPRS, 22.0 m × 5.0 m × 2.0 m) were studied. M. salmoides with initial an body weight of 8.25 ± 0.51 g and body length of 6.99 ± 0.44 cm were reared at an initial stocking density of 90.91 ind./m3 (low stocking density, LSD), 113.63 ind./m3 (middle stocking density, MSD), and 136.36 ind./m3 (high stocking density, HSD) with triplication. After 300 days of culture, MSD recorded the highest final body weight, weight gain, specific growth rate, and yield, but the food conversion ratio in MSD was the lowest. The viscerosomatic index in LSD was significantly higher than other groups. The fish serum reared at HSD showed significantly lower total protein, higher total cholesterol, triglyceride, total bilirubin, glucose content, alanine transaminase, and aspartate transaminase activity. Significantly lower intestinal amylase, lipase, trypsin activities, hepatic superoxide dismutase (SOD) and catalase (CAT) activities, and higher malondialdehyde content were detected in HSD compared to others. The content of crude lipid, saturated fatty acid decreased, and total essential amino acid, delicious amino acid, and polyunsaturated fatty acid increased in muscle with stocking density increase. No significant difference was observed in muscle texture. Profitability analysis indicated the benefit-to-cost ratio varied between 1.10 and 1.68, of which MSD was significantly higher than others. The optimal stocking density for M. salmoides should be 113.63 ind./m3 in an IPRS farm.


Assuntos
Aquicultura/métodos , Bass , Alanina Transaminase/sangue , Aminoácidos/metabolismo , Amilases/metabolismo , Animais , Aspartato Aminotransferases/sangue , Bass/sangue , Bass/crescimento & desenvolvimento , Bass/imunologia , Bass/metabolismo , Catalase/metabolismo , Ácidos Graxos/metabolismo , Proteínas de Peixes/sangue , Imunidade , Intestinos/enzimologia , Lipase/metabolismo , Fígado/metabolismo , Músculos/química , Esteróis/sangue , Superóxido Dismutase/metabolismo , Triglicerídeos/sangue , Tripsina/metabolismo
3.
Int J Mol Sci ; 22(14)2021 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-34299188

RESUMO

Na-K-ATPase provides a favorable transcellular Na gradient required for the functioning of Na-dependent nutrient transporters in intestinal epithelial cells. The primary metabolite for enterocytes is glutamine, which is absorbed via Na-glutamine co-transporter (SN2; SLC38A5) in intestinal crypt cells. SN2 activity is stimulated during chronic intestinal inflammation, at least in part, secondarily to the stimulation of Na-K-ATPase activity. Leukotriene D4 (LTD4) is known to be elevated in the mucosa during chronic enteritis, but the way in which it may regulate Na-K-ATPase is not known. In an in vitro model of rat intestinal epithelial cells (IEC-18), Na-K-ATPase activity was significantly stimulated by LTD4. As LTD4 mediates its action via Ca-dependent protein kinase C (PKC), Ca levels were measured and were found to be increased. Phorbol 12-myristate 13-acetate (PMA), an activator of PKC, also mediated stimulation of Na-K-ATPase like LTD4, while BAPTA-AM (Ca chelator) and calphostin-C (Cal-C; PKC inhibitor) prevented the stimulation of Na-K-ATPase activity. LTD4 caused a significant increase in mRNA and plasma membrane protein expression of Na-K-ATPase α1 and ß1 subunits, which was prevented by calphostin-C. These data demonstrate that LTD4 stimulates Na-K-ATPase in intestinal crypt cells secondarily to the transcriptional increase of Na-K-ATPase α1 and ß1 subunits, mediated via the Ca-activated PKC pathway.


Assuntos
Cálcio/metabolismo , Enterite/enzimologia , Células Epiteliais/enzimologia , Intestinos/enzimologia , Leucotrieno D4/farmacologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Animais , Sobrevivência Celular/fisiologia , Células Cultivadas , Enterite/tratamento farmacológico , Enterite/patologia , Ativação Enzimática , Células Epiteliais/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Proteína Quinase C/metabolismo , Ratos
4.
Life Sci ; 281: 119754, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34174323

RESUMO

AIM: The present study was undertaken to elucidate the potential protective mechanism of berberine (BBR) and/or zinc (Zn) against methotrexate (MTX)-induced intestinal injury. METHODS: Five groups of rats were assigned; normal group (received vehicle), MTX group (20 mg/kg; i.p. single dose), and the other three groups received a single daily oral dose of BBR (50 mg/kg), Zn (5 mg/kg), and BBR plus Zn respectively, for 5 days before MTX and 5 days after. RESULTS: Our results emphasized the toxic effect of MTX on rat's intestine as shown by disturbance of oxidant/antioxidant status, down-regulation of NRF2, SIRT1, FOXO-3, Akt, and mTOR expressions, along with up-regulation of GSK-3ß, JAK1, and STAT-3 expressions. Besides, severe intestinal histopathological changes were also observed. On the contrary, BBR and/or Zn produced marked protection against MTX-induced intestinal toxicity via amelioration of oxidative stress, improving NRF2, SIRT1, FOXO-3, GSK-3ß, Akt, mTOR, JAK1, and STAT-3 alterations. Moreover, our treatments significantly restored histopathological abnormalities. Interestingly, combination therapy of BBR plus Zn exhibited higher effectiveness than mono-therapy. SIGNIFICANCE: BBR plus Zn could be used as a novel therapy for the treatment of MTX-induced intestinal damage through modulation of GSK-3ß/NRF2, Akt/mTOR, JAK1/STAT-3, and SIRT1/FOXO-3 signaling pathways.


Assuntos
Berberina/farmacologia , Proteína Forkhead Box O3/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Intestinos/efeitos dos fármacos , Janus Quinase 1/metabolismo , Metotrexato/toxicidade , Fator 2 Relacionado a NF-E2/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Zinco/farmacologia , Animais , Inflamação/prevenção & controle , Intestinos/enzimologia , Intestinos/patologia , Masculino , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Sirtuína 1/metabolismo
5.
Arch Insect Biochem Physiol ; 107(3): e21792, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33948994

RESUMO

Although the importance of intestinal hydrolases is recognized, there is little information on the intestinal proteome of lepidopterans such as Anticarsia gemmatalis. Thus, we carried out the proteomic analysis of the A. gemmatalis intestine to characterize the proteases by LC/MS. We examined the interactions of proteins identified with protease inhibitors (PI) using molecular docking. We found 54 expressed antigens for intestinal protease, suggesting multiple important isoforms. The hydrolytic arsenal featured allows for a more comprehensive understanding of insect feeding. The docking analysis showed that the soybean PI (SKTI) could bind efficiently with the trypsin sequences and, therefore, insect resistance does not seem to involve changing the sequences of the PI binding site. In addition, a SERPIN was identified and the interaction analysis showed the inhibitor binding site is in contact with the catalytic site of trypsin, possibly acting as a regulator. In addition, this SERPIN and the identified PI sequences can be targets for the control of proteolytic activity in the caterpillar intestine and serve as a support for the rational design of a molecule with greater stability, less prone to cleavage by proteases and viable for the control of insect pests such as A. gemmatalis.


Assuntos
Mariposas/enzimologia , Peptídeo Hidrolases/metabolismo , Sequência de Aminoácidos , Animais , Intestinos/enzimologia , Larva/enzimologia , Simulação de Acoplamento Molecular , Mariposas/genética , Peptídeo Hidrolases/química , Peptídeo Hidrolases/genética
6.
J Insect Sci ; 21(3)2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33974083

RESUMO

Host plant preference of agricultural pests may shift throughout the growing season, allowing the pests to persist on wild hosts when crops are not available. Lygus Hahn (Hemiptera: Miridae) bugs are severe pests of cotton during flowering and fruiting stages, but can persist on alternative crops, or on weed species. Diversity of digestive enzymes produced by salivary glands and gut tissues play a pivotal role in an organism's ability to utilize various food sources. Polyphagous insects produce an array of enzymes that can process carbohydrates, lipids, and proteins. In this study, the digestive enzyme repertoire of the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), was identified by high-throughput sequencing followed by cDNA cloning and sequencing. This study identified 87 digestive genes, including 30 polygalacturonases (PG), one ß-galactosidase, three α-glucosidases, six ß-glucosidases, 28 trypsin-like proteases, three serine proteases, one apyrase-like protease, one cysteine protease, 12 lipases, and two transcripts with low similarity to a xylanase A-like genes. RNA-Seq expression profiles of these digestive genes in adult tarnished plant bugs revealed that 57 and 12 genes were differentially expressed in the salivary gland and gut (≥5-fold, P ≤ 0.01), respectively. All polygalacturonase genes, most proteases, and two xylanase-like genes were differentially expressed in salivary glands, while most of the carbohydrate and lipid processing enzymes were differentially expressed in the gut. Seven of the proteases (KF208689, KF208697, KF208698, KF208699, KF208700, KF208701, and KF208702) were not detected in either the gut or salivary glands.


Assuntos
Digestão/genética , Heterópteros , Intestinos/enzimologia , Glândulas Salivares/enzimologia , Transcriptoma , Animais , Genes de Insetos , Heterópteros/enzimologia , Heterópteros/genética , RNA-Seq/métodos
7.
Food Chem ; 361: 130047, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34029903

RESUMO

Inhibition of maltase, sucrase, isomaltase and glucoamylase activity by acarbose, epigallocatechin gallate, epicatechin gallate and four polyphenol-rich tea extract from white, green, oolong, black tea, were investigated by using rat intestinal enzymes and human Caco-2 cells. Regarding rat intestinal enzyme mixture, all four tea extracts were very effective in inhibiting maltase and glucoamylase activity, but only white tea extract inhibited sucrase and isomaltase activity and the inhibition was limited. Mixed-type inhibition on rat maltase activity was observed. Tea extracts in combination with acarbose, produced a synergistic inhibitory effect on rat maltase activity. Caco-2 cells experiments were conducted in Transwells. Green tea extract and epigallocatechin gallate show dose-dependent inhibition on human sucrase activity, but no inhibition on rat sucrase activity. The opposite was observed on maltase activity. The results highlighted the different response in the two investigated model systems and show that tea polyphenols are good inhibitors for α-glucosidase activity.


Assuntos
Glicosídeo Hidrolases/antagonistas & inibidores , Intestinos/enzimologia , Extratos Vegetais/química , Polifenóis/farmacologia , Chá/química , Acarbose/farmacologia , Animais , Células CACO-2 , Catequina/análogos & derivados , Catequina/farmacologia , Glucana 1,4-alfa-Glucosidase/antagonistas & inibidores , Inibidores de Glicosídeo Hidrolases/farmacologia , Humanos , Cinética , Oligo-1,6-Glucosidase/antagonistas & inibidores , Ratos , Sacarase/antagonistas & inibidores , alfa-Glucosidases/efeitos dos fármacos
8.
Mar Drugs ; 19(3)2021 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-33803803

RESUMO

Obesity is a complex metabolic disease, which is increasing worldwide. The reduction of dietary lipid intake is considered an interesting pathway to reduce fat absorption and to affect the chronic energy imbalance. In this study, zebrafish larvae were used to analyze effects of cyanobacteria on intestinal lipid absorption in vivo. In total, 263 fractions of a cyanobacterial library were screened for PED6 activity, a fluorescent reporter of intestinal lipases, and 11 fractions reduced PED6 activity > 30%. Toxicity was not observed for those fractions, considering mortality, malformations or digestive physiology (protease inhibition). Intestinal long-chain fatty acid uptake (C16) was reduced, but not short-chain fatty acid uptake (C5). Alteration of lipid classes by high-performance thin-layer chromatography (HPTLC) or lipid processing by fluorescent HPTLC was analyzed, and 2 fractions significantly reduced the whole-body triglyceride level. Bioactivity-guided feature-based molecular networking of LC-MS/MS data identified 14 significant bioactive mass peaks (p < 0.01, correlation > 0.95), which consisted of 3 known putative and 11 unknown compounds. All putatively identified compounds were known to be involved in lipid metabolism and obesity. Summarizing, some cyanobacterial strains repressed intestinal lipid absorption without any signs of toxicity and could be developed in the future as nutraceuticals to combat obesity.


Assuntos
Fármacos Antiobesidade/farmacologia , Cianobactérias/metabolismo , Inibidores Enzimáticos/farmacologia , Absorção Intestinal/efeitos dos fármacos , Intestinos/efeitos dos fármacos , Lipase/antagonistas & inibidores , Metabolismo dos Lipídeos/efeitos dos fármacos , Proteínas de Peixe-Zebra/antagonistas & inibidores , Peixe-Zebra/metabolismo , Animais , Fármacos Antiobesidade/isolamento & purificação , Inibidores Enzimáticos/isolamento & purificação , Intestinos/enzimologia , Lipase/metabolismo , Peixe-Zebra/embriologia , Proteínas de Peixe-Zebra/metabolismo
9.
Bioorg Chem ; 110: 104720, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33662896

RESUMO

Naturally occurring anti-diabetic compound curcumin can prevent diabetes complications due to antioxidant and anti-inflammatory properties as well as the attenuation of postprandial hyperglycemia. In this line, we have synthesized thirteen curcumin based derivatives (L1-L13) by multi-component reaction, characterized by IR, 1HNMR, 13C NMR, MS, elemental analysis and evaluated for possible antioxidant properties and α-glucosidase (α-Glu) and α-amylase (α-Amy) inhibitory potential. The curcumin-based pyrano[2,3-d]pyrimidine derivatives could inhibit α-Glu and α-Amy enzyme activity which showed desirable antioxidant activity. Furthermore, among the series, L5, L12, L9, L10, L8 and L11 were identified as more potent inhibitors of α-Glu enzyme than curcumin and the compounds of L12, L4, L9, L5, L10, L8, L13, and L11 were the stronger inhibitors of the α-Amy enzyme in vitro. Besides, among them, L12 had the lowest IC50 for the inhibition of both enzymes. Since strong inhibitors for pancreatic α-Amy result in the progression of severe gastrointestinal side effects, the inhibitors that show the lower α-Amy/α-Glu inhibitory ratio have attracted much attention in medicinal chemistry. Besides, considering antioxidant characteristics of synthesized compounds, the L7 derivative with the highest antioxidant activity and the lowest "α-Amy/α-Glu inhibitory" ratio could be an appropriate candidate for further study through the rational drug design to the exploration of a new class of powerful anti-diabetic drugs.


Assuntos
Antioxidantes/síntese química , Antioxidantes/farmacologia , Curcumina/química , Diabetes Mellitus/tratamento farmacológico , Glicosídeo Hidrolases/antagonistas & inibidores , Hipoglicemiantes/farmacologia , Animais , Desenho de Fármacos , Hipoglicemiantes/síntese química , Hipoglicemiantes/química , Intestinos/enzimologia , Modelos Moleculares , Simulação de Acoplamento Molecular , Estrutura Molecular , Pâncreas/enzimologia , Ligação Proteica , Conformação Proteica , Ratos , Relação Estrutura-Atividade , alfa-Amilases/genética , alfa-Amilases/metabolismo , alfa-Glucosidases/genética , alfa-Glucosidases/metabolismo
10.
Food Chem Toxicol ; 151: 112146, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33766611

RESUMO

Sustainable freshwater aquaculture has been recently gaining attention owing to the potential of nourishing the world. The study aimed to evaluate the influence of finishing diets on the activity of 21 genes involved in hepatic lipid metabolism and intestinal homeostasis, liver and intestine histology, and the level of eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids in common carp fillets. We compared two experimental diets: control diet mimicking a commercial feed (CTRL) and a test diet (CB) fortified with EPA and DHA retrieved from salmon by-products. An additional control (eCTRL) from extensively cultured carps was investigated. The study revealed that the expression of seven hepatic genes, e.g., lipoprotein lipase and fatty acid synthase, and six intestinal genes e.g., claudin-3c and γ-glutamyl transpeptidase, was influenced specifically by the experimental diets and farming type. Fish from the eCTRL group had the smallest hepatocytes and the largest nuclei compared with CTRL and CB. No pathological signs were found in intestine samples. Additionally, the levels of EPA and DHA in fillets were significantly higher in fish receiving CB compared with CTRL and eCTRL. The use of fortified diets is a promising solution to produce freshwater species with enhanced nutritional value without compromising the safety of fillets.


Assuntos
Ração Animal , Ácidos Graxos Ômega-3/administração & dosagem , Animais , Aquicultura , Carpas , Regulação da Expressão Gênica , Intestinos/enzimologia , Metabolismo dos Lipídeos , Fígado/enzimologia , Fígado/metabolismo
11.
Front Immunol ; 12: 635015, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33717179

RESUMO

An 8-week rearing trial was designed to appraise the dietary lysine levels on intestinal antioxidant capacity and immunity of grass carp fry. Six practical diets were prepared with graded levels of lysine (1.44, 1.79, 1.97, 2.44, 2.56 and 2.87% dry matter), and these diets were fed to grass carp fry. The results showed that the activities of intestinal antioxidant factors including catalase and glutathione peroxidase were markedly improved by the 2.44% dietary lysine compared with the control diet (1.44% dietary lysine) (P < 0.05). In terms of antioxidants, compared with the control diet, the 2.44% diet markedly upregulated the mRNA expression levels of target of rapamycin, S6 kinase1 and nuclear factor erythroid 2-related factor 2 pathway-related antioxidant genes, containing catalase and glutathione peroxidase 1α (P < 0.05) and downregulated the mRNA levels of Kelch-like ECH-associated protein 1 (P > 0.05). The mRNA levels of 4E-binding protein 2 showed the opposite trend compared with those of target of rapamycin, and the minimum value was observed in the group of 1.97% dietary lysine (P < 0.05). In terms of immunity, compared with the 1.44% diet, the 2.44% diet markedly suppressed the intestinal p38 mitogen-activated protein kinase and interferon γ2 mRNA levels (P < 0.05). Moreover, nuclear factor-kappa B p65, tumor necrosis factor α, interleukin 6, interleukin 8, and interleukin 15 mRNA levels all exhibited the same trend as p38 mitogen-activated protein kinase and interferon γ2; however, the difference among all the lysine treatments groups was not significant (P > 0.05). The anti-inflammatory cytokines transforming growth factor ß2 and interleukin 4/13B mRNA levels in the intestine were remarkably upregulated by high dietary lysine levels (2.56 and 2.87%) (P < 0.05), and when the dietary lysine level reached 2.44%, the interleukin 4/13A mRNA levels were strikingly increased (P < 0.05). Overall, the data suggested that 2.44% dietary lysine could strengthen the immune and antioxidant capacities of grass carp fry via activating the target of rapamycin (TOR) signaling pathway, and suppressing the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway, which then improve the survival rate.


Assuntos
Ração Animal , Antioxidantes/metabolismo , Carpas/metabolismo , Citocinas/metabolismo , Proteínas de Peixes/metabolismo , Intestinos/enzimologia , Lisina/administração & dosagem , Serina-Treonina Quinases TOR/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Animais , Carpas/genética , Carpas/imunologia , Citocinas/genética , Proteínas de Peixes/genética , Regulação da Expressão Gênica , Intestinos/imunologia , Lisina/metabolismo , Transdução de Sinais , Serina-Treonina Quinases TOR/genética , Proteínas Quinases p38 Ativadas por Mitógeno/genética
12.
Fish Physiol Biochem ; 47(4): 869-880, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33770301

RESUMO

The trials of finding non-conventional and alternative aquafeed ingredients are increasing. In this sense, this study evaluated the influence of coconut oil on the growth, feed utilization, immune, and antioxidative responses of Nile tilapia. Five test diets were formulated by mixing coconut oil with the other ingredients at 0, 1, 2, 3, and 4% of the total ration and presented for tilapia for 60 successive days. The final weight, SGR, weight gain (WG), and feed intake were superior in fish delivered 2% of coconut oil (P < 0.05). Concurrently, fish that received 2% coconut oil had lower FCR and higher PER than fish of the control and 4% groups (P < 0.05). Higher lipase activity was observed in fish of 2% and 3% levels than the remaining groups (P < 0.05). Besides, the amylase and protease activities of fish in 1%, 2%, and 3% groups were higher than the 0% level (P < 0.05). The total blood cholesterol, RBCs, and PCV showed higher values in Nile tilapia fed 2% and 3% coconut oil (P < 0.05). The lysozyme and phagocytic activities were higher in fish fed 2% and 3% levels than the control (P < 0.05), while the phagocytic index in 2% and 3% levels was higher than 0% and 4% levels. Furthermore, SOD and CAT were higher in fish fed 1%, 2%, and 3% than fish fed 0% and 4% levels while GSH was higher in fish of 1%, 2%, and 3% than fish fed 0% level (P < 0.05). However, the MDA level was markedly lower in fish fed 25, 3%, and 4% coconut oil than the 0% level (P < 0.05). The intestine's histological structure in all groups appeared normal, forming of intestinal villi projecting from the intestinal wall. Also, the structure of the hepatopancreas had a normal architecture in all groups. To sum up, the inclusion of coconut oil at 2 to 3% is recommended as a replacer for fish oil in Nile tilapia diets.


Assuntos
Ciclídeos , Óleo de Coco/farmacologia , Suplementos Nutricionais , Amilases/metabolismo , Animais , Antioxidantes , Aquicultura/métodos , Ciclídeos/anatomia & histologia , Ciclídeos/crescimento & desenvolvimento , Ciclídeos/imunologia , Ciclídeos/metabolismo , Hepatopâncreas/anatomia & histologia , Intestinos/anatomia & histologia , Intestinos/enzimologia , Lipase/metabolismo , Fígado/anatomia & histologia , Peptídeo Hidrolases/metabolismo , Fagossomos/efeitos dos fármacos , Fagossomos/fisiologia
13.
Proc Natl Acad Sci U S A ; 118(10)2021 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-33649233

RESUMO

Most animals harbor a gut microbiota that consists of potentially pathogenic, commensal, and mutualistic microorganisms. Dual oxidase (Duox) is a well described enzyme involved in gut mucosal immunity by the production of reactive oxygen species (ROS) that antagonizes pathogenic bacteria and maintains gut homeostasis in insects. However, despite its nonspecific harmful activity on microorganisms, little is known about the role of Duox in the maintenance of mutualistic gut symbionts. Here we show that, in the bean bug Riptortus pedestris, Duox-dependent ROS did not directly contribute to epithelial immunity in the midgut in response to its mutualistic gut symbiont, Burkholderia insecticola Instead, we found that the expression of Duox is tracheae-specific and its down-regulation by RNAi results in the loss of dityrosine cross-links in the tracheal protein matrix and a collapse of the respiratory system. We further demonstrated that the establishment of symbiosis is a strong oxygen sink triggering the formation of an extensive network of tracheae enveloping the midgut symbiotic organ as well as other organs, and that tracheal breakdown by Duox RNAi provokes a disruption of the gut symbiosis. Down-regulation of the hypoxia-responsive transcription factor Sima or the regulators of tracheae formation Trachealess and Branchless produces similar phenotypes. Thus, in addition to known roles in immunity and in the formation of dityrosine networks in diverse extracellular matrices, Duox is also a crucial enzyme for tracheal integrity, which is crucial to sustain mutualistic symbionts and gut homeostasis. We expect that this is a conserved function in insects.


Assuntos
Burkholderia/crescimento & desenvolvimento , Oxidases Duais/metabolismo , Heterópteros , Proteínas de Insetos/metabolismo , Intestinos , Simbiose/fisiologia , Animais , Oxidases Duais/genética , Heterópteros/enzimologia , Heterópteros/genética , Heterópteros/microbiologia , Proteínas de Insetos/genética , Intestinos/enzimologia , Intestinos/microbiologia
14.
Food Chem ; 354: 129513, 2021 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-33765464

RESUMO

A raw starch digesting α-amylase from Nile tilapia (Oreochromis niloticus) intestine was identified. The α-amylase, AMY-T, had an estimated molecular weight of 60 kDa and purified to near homogeneity. AMY-T showed an apparent KM 4.78 mg/mL and Vmax 0.44 mg/mL/min) towards soluble starch. It was highly stable for 24 h in the pH range 3.0-10.0, and to solvents like methanol, isopropanol, butanol, dimethylformamide, DMSO and ethyl-ether. AMY-T was able to digest different carbohydrates, mainly showing endo-activity. Importantly, AMY-T was catalytically efficient and adsorbing towards raw potato starch at temperature documented for other raw starch digesting α-amylases. Thin layer and anion exchange chromatography characterization showed that the end products of raw starch hydrolysis were glucose, maltose and maltodextrins, with degree of polymerisation ranging 1-8. Scanning electron microscopy analysis of the AMY-T treated starch granules documented both granular exo- and endo-attack by AMY-T. These catalytic capabilities suggest high potential for AMY-T for industrial use.


Assuntos
Ciclídeos/metabolismo , Intestinos/enzimologia , Amido/metabolismo , alfa-Amilases/metabolismo , Animais , Cromatografia por Troca Iônica , Cromatografia em Camada Delgada , Concentração de Íons de Hidrogênio , Hidrólise , Maltose/análise , Polissacarídeos/análise , Amido/análise , Temperatura
15.
J Ethnopharmacol ; 271: 113914, 2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-33571617

RESUMO

ETHNOPHARMACOLOGICAL RELEVANCE: Shengmai San (SMS) has been commonly used as a traditional Chinese medicine for the treatment of cardiovascular disorders, of which drug interactions need to be assessed for the safety concern. There is little evidence for the alterations of hepatic and intestinal drug-metabolizing enzymes after repeated SMS treatments to assess drug interactions. AIM OF THE STUDY: The studies aim to illustrate the effects of repeated treatments with SMS on cytochrome P450s (CYPs), reduced nicotinamide adenine dinucleotide (phosphate)-quinone oxidoreductase (NQO), uridine diphosphate-glucuronosyltransferase (UGT), and glutathione S-transferase (GST) using in vivo rat model. MATERIALS AND METHODS: The SMS was prepared using Schisandrae Fructus, Ginseng Radix, and Ophiopogonis Radix (OR) (1:2:2). Chromatographic analyses of decoctions were performed using ultra-performance liquid chromatography (UPLC) and LC-mass spectrometry. Sprague-Dawley rats were orally treated with the SMS and its component herbal decoctions for 2 or 3 weeks. Hepatic and intestinal enzyme activities were determined. CYP3A expression and the kinetics of intestinal nifedipine oxidation (NFO, a CYP3A marker reaction) were determined. RESULTS: Schisandrol A, schisandrin B, ginsenoside Rb1 and ophiopogonin D were identified in SMS. SMS selectively suppressed intestinal, but not hepatic, NFO activity in a dose- and time-dependent manner. Hepatic and intestinal UGT, NQO and GST activities were not affected. A 3-week SMS treatment decreased the maximal velocity of intestinal NFO by 50%, while the CYP3A protein level remained unchanged. Among SMS component herbs, the decoction of OR decreased intestinal NFO activity. CONCLUSIONS: These findings demonstrate that 3-week treatment with SMS and OR suppress intestinal, but not hepatic CYP3A function. It suggested that the potential interactions of SMS with CYP 3A drug substrates should be noticed, especially the drugs whose bioavailability depends heavily on intestinal CYP3A.


Assuntos
Inibidores do Citocromo P-450 CYP3A/farmacologia , Medicamentos de Ervas Chinesas/farmacologia , Intestinos/enzimologia , Fígado/enzimologia , Animais , Biomarcadores/sangue , Ciclo-Octanos/análise , Citocromo P-450 CYP3A/metabolismo , Inibidores do Citocromo P-450 CYP3A/análise , Inibidores do Citocromo P-450 CYP3A/uso terapêutico , Combinação de Medicamentos , Medicamentos de Ervas Chinesas/análise , Medicamentos de Ervas Chinesas/uso terapêutico , Ginsenosídeos/análise , Glucuronosiltransferase/metabolismo , Glutationa Transferase/metabolismo , Interações Ervas-Drogas , Intestinos/efeitos dos fármacos , Lignanas/análise , Fígado/efeitos dos fármacos , Masculino , Microssomos/efeitos dos fármacos , Microssomos/enzimologia , NAD(P)H Desidrogenase (Quinona)/metabolismo , Nifedipino/metabolismo , Oxirredução/efeitos dos fármacos , Compostos Policíclicos/análise , Ratos Sprague-Dawley , Saponinas/química , Espirostanos/química
16.
Toxicol In Vitro ; 72: 105087, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33440186

RESUMO

UDP glucuronosyltransferases (UGTs) of the gastrointestinal tract play a crucial role in protection against the toxic effects of xenobiotics in the environment. UGTs such as UGT1A8 and UGT1A10 are predominantly expressed in gastrointestinal tissues. In this study, we examined the phase II metabolism of raloxifene in differentiated Caco-2 monolayers by inducing UGT1A8 and UGT1A10 expression in these cells. The present study evaluated the following four flavonoids of Scutellaria baicalensis as model herbal compounds: scutellarein, salvigenin, baicalein, and wogonin. All test compounds, endpoint substrates, and their metabolites were quantified using liquid chromatography and high-resolution mass spectrometry. The transepithelial electrical resistance values for the individual compounds were comparable regardless of whether they were measured individually. Salvigenin significantly inhibited UGT1A8 and UGT1A10 activities in a concentration-dependent manner. All individual compounds except scutellarein inhibited UGT1A8 and UGT1A10 activity at a concentration of 100 µM. In addition, all individual flavonoids at 100 µM, except wogonin, significantly increased the amount of raloxifene in the basolateral chambers. The positive control, canagliflozin, significantly inhibited both UGT1A8 and UGT1A10 activities. These findings suggest that the Caco-2 assay can be utilized for identifying UGT1A8 and UGT1A10 inhibitors and indicate the potential of salvigenin for enhancing the pharmacological effects of UGT substrate drugs.


Assuntos
Flavonoides/farmacologia , Glucuronosiltransferase/antagonistas & inibidores , Interações Ervas-Drogas , Cloridrato de Raloxifeno/farmacologia , Scutellaria baicalensis , Moduladores Seletivos de Receptor Estrogênico/farmacologia , Células CACO-2 , Glucuronídeos/metabolismo , Glucuronosiltransferase/metabolismo , Humanos , Intestinos/enzimologia
17.
Toxicol Lett ; 341: 1-10, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33429014

RESUMO

Piscine cytochrome P450 (CYP) enzymes play an important role in the metabolism of xenobiotics. Xenobiotics often act as inducers of CYP1A1 and CYP3A expression and activity in fish. We compared constitutive mRNA expression of CYP1A1, CYP3A27, and CYP3A45 and catalytic activity of CYP1A (7-ethoxyresorufin-O-deethylation, EROD) and CYP3A-like (benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylation, BFCOD) enzymes in the following six rainbow trout tissues: liver, gill, heart, brain, intestine, and gonad. mRNA expression and activity were present in all investigated tissues. The CYP1A1 mRNA expression was higher in the liver, gill, heart, and brain compared to gonad and intestine. The intestine was the main site of CYP3A27 and CYP3A45 expression. The highest EROD and BFCOD activity was observed in liver tissue followed in descending order by heart, brain, gill, intestine, and gonad. Such differences might be related to the role of CYP physiological functions in the specific tissue. Rainbow trout exposure to 50 mg/kg of ß-naphthoflavone for 48 h resulted in a 7.5- and 5.9-fold increase in liver EROD and BFCOD activity, respectively. In vitro EROD activity inhibition with ellipticine showed tissue-specific inhibition, while ketoconazole decreased BFCOD activity by 50-98 % in all tissues. Further studies are needed to identify all CYP isoforms that are responsible for these activities and modes of regulation.


Assuntos
Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP3A/metabolismo , Regulação Enzimológica da Expressão Gênica/fisiologia , Oncorhynchus mykiss/metabolismo , Animais , Encéfalo/enzimologia , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP3A/genética , Feminino , Brânquias/enzimologia , Intestinos/enzimologia , Fígado/enzimologia , Masculino , Miocárdio/enzimologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Caracteres Sexuais
18.
PLoS One ; 16(1): e0245649, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33471847

RESUMO

Rapid adaptive responses were evident from reciprocal host-plant switches on performance, digestive physiology and relative gene expression of gut serine proteases in larvae of crucifer pest P. brassicae transferred from cauliflower (CF, Brassica oleracea var. botrytis, family Brassicaceae) to an alternate host, garden nasturtium, (GN, Tropaeolum majus L., family Tropaeolaceae) and vice-versa under laboratory conditions. Estimation of nutritional indices indicated that larvae of all instars tested consumed the least food and gained less weight on CF-GN diet (significant at p≤0.05) as compared to larvae feeding on CF-CF, GN-GN and GN-CF diets suggesting that the switch to GN was nutritionally less favorable for larval growth. Nevertheless, these larvae, especially fourth instars, were adroit in utilizing and digesting GN as a new host plant type. In vitro protease assays conducted to understand associated physiological responses within twelve hours indicated that levels and properties of gut proteases were significantly influenced by type of natal host-plant consumed, change in diet as well as larval age. Activities of gut trypsins and chymotrypsins in larvae feeding on CF-GN and GN-CF diets were distinct, and represented shifts toward profiles observed in larvae feeding continuously on GN-GN and CF-CF diets respectively. Results with diagnostic protease inhibitors like TLCK, STI and SBBI in these assays and gelatinolytic zymograms indicated complex and contrasting trends in gut serine protease activities in different instars from CF-GN diet versus GN-CF diet, likely due to ingestion of plant protease inhibitors present in the new diet. Cloning and sequencing of serine protease gene fragments expressed in gut tissues of fourth instar P. brassicae revealed diverse transcripts encoding putative trypsins and chymotrypsins belonging to at least ten lineages. Sequences of members of each lineage closely resembled lepidopteran serine protease orthologs including uncharacterized transcripts from Pieris rapae. Differential regulation of serine protease genes (Pbr1-Pbr5) was observed in larval guts of P. brassicae from CF-CF and GN-GN diets while expression of transcripts encoding two putative trypsins (Pbr3 and Pbr5) were significantly different in larvae from CF-GN and GN-CF diets. These results suggested that some gut serine proteases that were differentially expressed in larvae feeding on different species of host plants were also involved in rapid adaptations to dietary switches. A gene encoding nitrile-specifier protein (nsp) likely involved in detoxification of toxic products from interactions of ingested host plant glucosinolates with myrosinases was expressed to similar levels in these larvae. Taken together, these snapshots reflected contrasts in physiological and developmental plasticity of P. brassicae larvae to nutritional challenges from wide dietary switches in the short term and the prominent role of gut serine proteases in rapid dietary adaptations. This study may be useful in designing novel management strategies targeting candidate gut serine proteases of P. brassicae using RNA interference, gene editing or crops with transgenes encoding protease inhibitors from taxonomically-distant host plants.


Assuntos
Comportamento Alimentar , Regulação Enzimológica da Expressão Gênica , Proteínas de Insetos/biossíntese , Intestinos/enzimologia , Lepidópteros/enzimologia , Serina Proteases/biossíntese , Animais , Proteínas de Insetos/genética , Larva/enzimologia , Larva/genética , Lepidópteros/genética , Serina Proteases/genética
19.
Int J Biol Macromol ; 171: 150-157, 2021 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-33418039

RESUMO

The cytochrome P450 monooxygenases of insects play crucial roles in the metabolic detoxification of insecticides. Our previous finding showed that two cytochrome P450 genes, both CYP301B1 and CYP6AX1v2, in the BPH underwent overexpression due to ß-asarone. In this study, we investigated the molecular characteristics, expression patterns and functions of these two cytochrome P450 genes. The results showed that CYP301B1 had the highest expression level in the eggs, while CYP6AX1v2 was expressed in macropterous female adults. Moreover, the expression level of CYP301B1 in the head was higher than that in the integument, fat body and gut. The expression level of CYP6AX1v2 in the fat body and gut was higher than that in head and integument. Importantly, silencing CYP301B1 and CYP6AX1v2 separately could increase the sensitivity, resulting in significant higher mortality of BPH following treatment with ß-asarone. Our findings indicated that CYP301B1 and CYP6AX1v2 could contribute to the resistance of BPH to ß-asarone, and these two genes may be involved in the detoxification metabolism of ß-asarone in BPH.


Assuntos
Anisóis/farmacologia , Sistema Enzimático do Citocromo P-450/genética , Hemípteros/efeitos dos fármacos , Inativação Metabólica/genética , Proteínas de Insetos/genética , Inseticidas/farmacologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Sistema Enzimático do Citocromo P-450/metabolismo , Corpo Adiposo/efeitos dos fármacos , Corpo Adiposo/enzimologia , Regulação da Expressão Gênica , Cabeça , Hemípteros/enzimologia , Hemípteros/genética , Proteínas de Insetos/antagonistas & inibidores , Proteínas de Insetos/metabolismo , Intestinos/efeitos dos fármacos , Intestinos/enzimologia , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Zigoto/efeitos dos fármacos , Zigoto/enzimologia
20.
J Fish Biol ; 98(3): 643-654, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33124694

RESUMO

The family Mugilidae consists mainly of diadromous species, whose reproduction occurs in offshore waters. Pre-juveniles shift their diet in the surf zone (zooplanktophagous to iliophagous). Later, during their recruitment into estuaries, huge changes take place in their digestive system. However, digestive and metabolic characteristics and some morphological traits at recruitment are unknown for Mugilidae. We performed comparative studies on early and late pre-juveniles of Mugil liza recruited in Mar Chiquita Coastal Lagoon (37°32'-37°45'S, 57°19'-57°26'W, Argentina). We determined digestive enzyme activities (intestine), energy reserves (liver/muscle), total/standard length, total weight, intestinal coefficient, hepatosomatic index and retroperitoneal fat. Pre-juveniles exhibited amylase, maltase, sucrase, lipase, trypsin and aminopeptidase-N (APN) activities, which were maintained over a wide range of pH and temperature, and exhibited Michaelis-Menten kinetics. In late pre-juveniles, amylase (422 ± 131 µmol maltose min-1 mgprot-1 ), sucrase (86 ± 14 mg glucose min-1 mgprot-1 ), trypsin (84 ± 9 µmoles min-1 mgprot-1 ) and APN (0.58 ± 0.08 µmoles min-1 mgprot-1 ) activities were higher (42%, 28%, 35% and 28%, respectively) than in the early stage. Also, the intestinal coefficient was higher in late (3.04) compared to early (2.06) pre-juveniles. Moreover, the liver appeared to be a main site of glycogen and triglyceride storage in late pre-juveniles, muscle being the site of storage in early pre-juveniles, exhibiting higher glycogen, free glucose and protein concentrations (92%, 82%, 32%, respectively). The results suggest that pre-juveniles of M. liza exhibit an adequate digestive battery to perform complete hydrolysis of various dietary substrates, availability of energy reserves and morphological characteristics to support their feeding habit and growth after recruitment. Our results represent an important contribution to knowledge of the ecology and digestive physiology of pre-juveniles of Mugilidae in the wild.


Assuntos
Digestão/fisiologia , Metabolismo Energético , Metaboloma , Smegmamorpha/crescimento & desenvolvimento , Smegmamorpha/metabolismo , Animais , Argentina , Dieta , Enzimas/metabolismo , Estuários , Glicogênio/metabolismo , Intestinos/enzimologia , Fígado/metabolismo , Músculos/metabolismo , Smegmamorpha/anatomia & histologia
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...