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1.
Pol J Pathol ; 70(3): 226-231, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31820869

RESUMO

Sertoli Leydig cell tumor of the ovary, is a rare neoplasm from the group of sex cord-stromal tumors of the ovary, accounting for less than 1% of all ovarian tumors. Among the Sertoli Leydig cell tumors, we distinguish a separate group of tumors secreting α-fetoprotein (AFP). The young 24-year-old woman presented to the Clinical Department of Gynaecological Endocrinology at the University Hospital in Krakow due to secondary amenorrhea, hirsutism and worsening abdominal pain for several months. During the admission draws attention was drawn to the abnormal level of testosterone, AFP and the revised structure of the ovary in the ultrasound. After a preliminary diagnosis, expanded pelvic MRI was performed, which found an isolated tumor derived from Sertoli Leydig cells. The patient was enrolled to unilaterally remove the right ovary by laparotomy. Histopathological examination and immunohistochemical staining confirmed the diagnosis of Sertoli Leydig cells tumor, and in pathological examination we found glandular mucosa cells of the colon. Owing to scientific reports on the stromal tumors of the ovary, we decided to perform genetic testing and verify the patient's karyotype. In the follow-up 90 days after the surgery, levels of testosterone and AFP were correct. In case of Sertoli Leydig cell tumors, especially in young women of childbearing potential, special attention should be paid to Anti-Mullerian hormone testing before surgery, as well as genetic diagnostics to exclude disorders of sex development.


Assuntos
Células Epiteliais/metabolismo , Neoplasias Ovarianas/patologia , Tumor de Células de Sertoli-Leydig/patologia , alfa-Fetoproteínas/metabolismo , Feminino , Humanos , Intestinos/citologia , Adulto Jovem
2.
J Agric Food Chem ; 67(49): 13758-13766, 2019 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-31789514

RESUMO

Probiotics, such as Lactobacillus, have been proven to be effective in maintaining intestinal homeostasis. The modulatory effect of Lactobacillus on intestinal epithelial development in early life is still unclear. In this study, Lactobacillus isolates with good probiotic abilities were screened and orally administered to detect their regulatory effect on intestinal development in chickens. L. reuteri 22 was isolated from chickens and chosen for subsequent chicken experiments due to its strong acid and bile salt resistance and ability to adhere to epithelial cells. The 3-day-old chickens were orally administrated with 108 CFU L. reuteri 22 for consecutive 7 days. L. reuteri 22 increased Lgr5 mRNA expression (3.23 ± 0.40, P = 0.001) and activated the Wnt/ß-catenin signaling pathway, with increasing expression of proliferating cell nuclear antigen (PCNA) (49.27 ± 9.81, P = 0.021) to support the proliferation of chicken intestinal epithelial cells. Moreover, L. reuteri 22 also inhibited the Notch signaling pathway to induce intestinal stem cell differentiation into goblet cells with increased mucin 2 (Muc-2) expression (1.72 ± 0.34, P = 0.047). L. reuteri 22 significantly enhanced lysozyme mRNA expression (2.32 ± 0.55, P = 0.019) to improve intestinal innate mucosal immunity. This study demonstrated that L. reuteri administration could regulate chicken intestinal epithelium development to ensure the function of the intestinal mucosal barrier, which is beneficial for newborn animals.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Epiteliais/citologia , Células Caliciformes/efeitos dos fármacos , Lactobacillus reuteri/fisiologia , Probióticos/farmacologia , Animais , Galinhas , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Células Caliciformes/citologia , Células Caliciformes/metabolismo , Intestinos/citologia , Intestinos/efeitos dos fármacos , Mucina-2/genética , Mucina-2/metabolismo , Antígeno Nuclear de Célula em Proliferação/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Transdução de Sinais/efeitos dos fármacos , beta Catenina/genética , beta Catenina/metabolismo
3.
Nature ; 574(7776): 112-116, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31554966

RESUMO

Organogenesis is a complex and interconnected process that is orchestrated by multiple boundary tissue interactions1-7. However, it remains unclear how individual, neighbouring components coordinate to establish an integral multi-organ structure. Here we report the continuous patterning and dynamic morphogenesis of hepatic, biliary and pancreatic structures, invaginating from a three-dimensional culture of human pluripotent stem cells. The boundary interactions between anterior and posterior gut spheroids differentiated from human pluripotent stem cells enables retinoic acid-dependent emergence of hepato-biliary-pancreatic organ domains specified at the foregut-midgut boundary organoids in the absence of extrinsic factors. Whereas transplant-derived tissues are dominated by midgut derivatives, long-term-cultured microdissected hepato-biliary-pancreatic organoids develop into segregated multi-organ anlages, which then recapitulate early morphogenetic events including the invagination and branching of three different and interconnected organ structures, reminiscent of tissues derived from mouse explanted foregut-midgut culture. Mis-segregation of multi-organ domains caused by a genetic mutation in HES1 abolishes the biliary specification potential in culture, as seen in vivo8,9. In sum, we demonstrate that the experimental multi-organ integrated model can be established by the juxtapositioning of foregut and midgut tissues, and potentially serves as a tractable, manipulatable and easily accessible model for the study of complex human endoderm organogenesis.


Assuntos
Sistema Biliar/embriologia , Intestinos/embriologia , Fígado/embriologia , Modelos Biológicos , Morfogênese , Pâncreas/embriologia , Animais , Sistema Biliar/citologia , Biomarcadores/análise , Biomarcadores/metabolismo , Padronização Corporal , Endoderma/citologia , Endoderma/embriologia , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/metabolismo , Intestinos/citologia , Fígado/citologia , Masculino , Camundongos , Organoides/citologia , Organoides/embriologia , Pâncreas/citologia , Esferoides Celulares/citologia , Esferoides Celulares/metabolismo , Esferoides Celulares/transplante , Fatores de Transcrição HES-1/análise , Fatores de Transcrição HES-1/metabolismo
4.
Nat Commun ; 10(1): 4123, 2019 09 11.
Artigo em Inglês | MEDLINE | ID: mdl-31511511

RESUMO

In adult epithelial stem cell lineages, the precise differentiation of daughter cells is critical to maintain tissue homeostasis. Notch signaling controls the choice between absorptive and entero-endocrine cell differentiation in both the mammalian small intestine and the Drosophila midgut, yet how Notch promotes lineage restriction remains unclear. Here, we describe a role for the transcription factor Klumpfuss (Klu) in restricting the fate of enteroblasts (EBs) in the Drosophila intestine. Klu is induced in Notch-positive EBs and its activity restricts cell fate towards the enterocyte (EC) lineage. Transcriptomics and DamID profiling show that Klu suppresses enteroendocrine (EE) fate by repressing the action of the proneural gene Scute, which is essential for EE differentiation. Loss of Klu results in differentiation of EBs into EE cells. Our findings provide mechanistic insight into how lineage commitment in progenitor cell differentiation can be ensured downstream of initial specification cues.


Assuntos
Linhagem da Célula , Proteínas de Ligação a DNA/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Enterócitos/citologia , Intestinos/citologia , Células-Tronco/citologia , Fatores de Transcrição/metabolismo , Animais , Carcinogênese/metabolismo , Carcinogênese/patologia , Diferenciação Celular , Proliferação de Células , Modelos Biológicos , Ligação Proteica , Receptores Notch/metabolismo , Transdução de Sinais , Células-Tronco/metabolismo
5.
Nature ; 574(7777): 254-258, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31534216

RESUMO

Group 3 innate lymphoid cells (ILC3s) are major regulators of inflammation, infection, microbiota composition and metabolism1. ILC3s and neuronal cells have been shown to interact at discrete mucosal locations to steer mucosal defence2,3. Nevertheless, it is unclear whether neuroimmune circuits operate at an organismal level, integrating extrinsic environmental signals to orchestrate ILC3 responses. Here we show that light-entrained and brain-tuned circadian circuits regulate enteric ILC3s, intestinal homeostasis, gut defence and host lipid metabolism in mice. We found that enteric ILC3s display circadian expression of clock genes and ILC3-related transcription factors. ILC3-autonomous ablation of the circadian regulator Arntl led to disrupted gut ILC3 homeostasis, impaired epithelial reactivity, a deregulated microbiome, increased susceptibility to bowel infection and disrupted lipid metabolism. Loss of ILC3-intrinsic Arntl shaped the gut 'postcode receptors' of ILC3s. Strikingly, light-dark cycles, feeding rhythms and microbial cues differentially regulated ILC3 clocks, with light signals being the major entraining cues of ILC3s. Accordingly, surgically or genetically induced deregulation of brain rhythmicity led to disrupted circadian ILC3 oscillations, a deregulated microbiome and altered lipid metabolism. Our work reveals a circadian circuitry that translates environmental light cues into enteric ILC3s, shaping intestinal health, metabolism and organismal homeostasis.


Assuntos
Encéfalo/efeitos da radiação , Ritmo Circadiano/efeitos da radiação , Homeostase/efeitos da radiação , Intestinos/imunologia , Intestinos/efeitos da radiação , Luz , Linfócitos/imunologia , Linfócitos/efeitos da radiação , Fatores de Transcrição ARNTL/deficiência , Fatores de Transcrição ARNTL/genética , Fatores de Transcrição ARNTL/metabolismo , Animais , Relógios Biológicos/genética , Relógios Biológicos/efeitos da radiação , Encéfalo/fisiologia , Ritmo Circadiano/genética , Ritmo Circadiano/imunologia , Ritmo Circadiano/fisiologia , Sinais (Psicologia) , Comportamento Alimentar/efeitos da radiação , Feminino , Microbioma Gastrointestinal/efeitos da radiação , Imunidade Inata/efeitos da radiação , Intestinos/citologia , Metabolismo dos Lipídeos , Linfócitos/metabolismo , Masculino , Camundongos , Fotoperíodo
6.
Nat Commun ; 10(1): 4066, 2019 09 06.
Artigo em Inglês | MEDLINE | ID: mdl-31492846

RESUMO

Human enteroviruses (HEVs) of the family Picornaviridae, which comprises non-enveloped RNA viruses, are ubiquitous worldwide. The majority of EV proteins are derived from viral polyproteins encoded by a single open reading frame (ORF). Here, we characterize a second ORF in HEVs that is crucial for viral intestinal infection. Disruption of ORF2p expression decreases the replication capacity of EV-A71 in human intestinal epithelial cells (IECs). Ectopic expression of ORF2p proteins derived from diverse enteric enteroviruses sensitizes intestinal cells to the replication of ORF2p-defective EV-A71 and respiratory enterovirus EV-D68. We show that the highly conserved WIGHPV domain of ORF2p is important for ORF2p-dependent viral intestinal infection. ORF2p expression is required for EV-A71 particle release from IECs and can support productive EV-D68 infection in IECs by facilitating virus release. Our results indicate that ORF2p is a determining factor for enteric enterovirus replication in IECs.


Assuntos
Enterovirus/genética , Fases de Leitura Aberta/genética , Vírus de RNA/genética , Replicação Viral/genética , Sequência de Aminoácidos , Sequência de Bases , Enterovirus/fisiologia , Infecções por Enterovirus/transmissão , Infecções por Enterovirus/virologia , Células Epiteliais/virologia , Fezes/virologia , Células HT29 , Interações Hospedeiro-Patógeno/genética , Humanos , Intestinos/citologia , Intestinos/virologia , Vírus de RNA/fisiologia , Proteínas Virais/genética , Proteínas Virais/metabolismo
7.
Vet Microbiol ; 235: 270-279, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31383312

RESUMO

Lawsonia intracellularis is an obligate intracellular microorganism and the causative agent of porcine proliferative enteropathy. Due to its obligate intracellular nature, characterization of antigens and proteins involved in host-pathogen interaction and immune recognition have been difficult to achieve using conventional microbiological techniques. In this work, we used 2-dimensional gel electrophoresis coupled with Western-immunoblotting, mass spectrometry and bioinformatics to identify bacterial proteins that interact in vitro with pig intestinal cells (IPEC-1), have immunogenic properties and the potential to be used as subunit vaccine antigens. We detected eleven immunogenic bacterial proteins from which fliC (LI0710), LI1153 (annotated by NCBI as Putative protein N), and LI0649 (annotated as autotransporter) were predicted to be expressed on the outer membrane while LI0169 (oppA; annotated as ABC dipeptide transport system) was predicted to be periplasmic with a transmembrane domain forming a central pore through the plasma membrane. Genes coding for these four proteins were cloned and expressed in Escherichia coli and the corresponding recombinant proteins were purified using affinity chromatography. Porcine hyperimmune serum against whole Lawsonia lysate established that all four recombinant proteins were immunogenic. Further, rabbit hyperimmune sera generated against the vaccine strain of L. intracellularis and rabbit serum specific for each recombinant protein showed an inhibitory effect on the attachment and penetration of live, avirulent L. intracellularis, thus indicating that each protein is a potential neutralizing antibody target and a candidate for subunit vaccine formulation.


Assuntos
Anticorpos Antibacterianos/imunologia , Anticorpos Neutralizantes/imunologia , Proteínas de Bactérias/imunologia , Vacinas Bacterianas/imunologia , Infecções por Desulfovibrionaceae/veterinária , Lawsonia (Bactéria)/imunologia , Animais , Proteínas de Bactérias/genética , Western Blotting , Linhagem Celular , Biologia Computacional , Infecções por Desulfovibrionaceae/imunologia , Infecções por Desulfovibrionaceae/prevenção & controle , Feminino , Intestinos/citologia , Intestinos/microbiologia , Espectrometria de Massas , Proteômica , Coelhos , Proteínas Recombinantes/imunologia , Suínos , Doenças dos Suínos/imunologia , Doenças dos Suínos/microbiologia , Doenças dos Suínos/prevenção & controle , Vacinas de Subunidades/imunologia
8.
Molecules ; 24(17)2019 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-31438521

RESUMO

Many studies have shown that Orthosiphon stamineus extract (OE) has antioxidant activity, and we previously reported that OE protects the intestine against injury from a high-fat diet. However, the molecular mechanism underlying this protective effect of OE was unclear. Here, OE was separated according to polarity and molecular weight, and the antioxidant activity of each component was compared. The components with the highest antioxidant activity were analyzed by HPLC, which confirmed that rosmarinic acid (RA) was the main effective constituent in OE. OE and RA were then tested in a mouse high-fat diet-induced intestinal injury model. The antioxidant indices and morphological characteristics of the mouse jejunum were measured, and activation of the nuclear factor E2-related factor 2 (Nrf2) pathway and apoptosis of jejunal epithelial cells were analyzed. Of all the constituents in OE, RA contributed the most. Both RA and OE activated the Nrf2 pathway and increased downstream antioxidant enzyme activity. RA and OE protected the mouse intestine against high-fat diet-induced oxidative stress by preventing intestinal epithelial cell apoptosis via both extracellular and intracellular pathways. Thus, RA, the main effective constituent in OE, inhibits intestinal epithelial apoptosis by regulating the Nrf2 pathway in mice.


Assuntos
Cinamatos/farmacologia , Depsídeos/farmacologia , Fator 2 Relacionado a NF-E2/metabolismo , Orthosiphon/química , Animais , Apoptose/efeitos dos fármacos , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/metabolismo , Intestinos/citologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo
9.
Org Biomol Chem ; 17(32): 7497-7506, 2019 08 28.
Artigo em Inglês | MEDLINE | ID: mdl-31365007

RESUMO

The nature and coordination sites of the Schiff base 3,3'-(1E,1'E)-(1,3-phenylenebis(azan-1-yl-1-ylidene))bis(methan-1-yl-1-ylidene)dinaphthalen-2-ol (APHN) were tuned by its selective reduction to design a highly efficient fluorescent probe, 3,3'-(pyridine-2,6-diylbis(azanediyl))bis(methylene)dinaphthalen-2-ol (RAPHN). The structures of APHN, RAPHN, and the RAPHN-Fe3+ complex were satisfactorily modeled from the results of density functional theory (DFT) and time-dependent DFT (TD-DFT) calculations. RAPHN worked in pure aqueous medium as a turn on-off-on probe of Fe3+ and F-. The fluorescence nature of the probe in the presence and absence of Fe3+/F- was regulated by a set of mechanisms including -CH[double bond, length as m-dash]N isomerization and LMCT. A 2 : 1 (M : L) binding stoichiometry was established from a fluorescence Job's plot and further substantiated from HR-MS studies. The limits of detection of RAPHN for Fe3+ and RAPHN-Fe3+ for F- were found to be 2.49 × 10-7 M and 1.09 × 10-7 M, respectively. The RAPHN probe caused no cytotoxicity in gut tissue of Drosophila even at high concentrations. The probe displayed excellent bioimaging applications for detection of Fe3+ and F- in gut tissue of Drosophila. A combinatorial logic gate was constructed for the proper understanding of the working principle of RAPHN.


Assuntos
Complexos de Coordenação/química , Corantes Fluorescentes/química , Íons/análise , Bases de Schiff/química , Animais , Técnicas Biossensoriais , Linhagem Celular , Teoria da Densidade Funcional , Drosophila , Intestinos/citologia , Cinética , Ligantes , Modelos Moleculares , Naftalenos/química , Espectrometria de Fluorescência
10.
Nat Commun ; 10(1): 2988, 2019 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-31278345

RESUMO

Precise control of stem cell (SC) proliferation ensures tissue homeostasis. In the Drosophila intestine, injury-induced regeneration involves initial activation of intestinal SC (ISC) proliferation and subsequent return to quiescence. These two phases of the regenerative response are controlled by differential availability of the BMP type I receptor Thickveins (Tkv), yet how its expression is dynamically regulated remains unclear. Here we show that during homeostasis, the E3 ubiquitin ligase Highwire and the ubiquitin-proteasome system maintain low Tkv protein expression. After ISC activation, Tkv is stabilized by proteasome inhibition and undergoes endocytosis due to the induction of the nucleoside diphosphate kinase Abnormal Wing Disc (AWD). Tkv internalization is required for the activation of the Smad protein Mad, and for the return to quiescence after a regenerative episode. Our data provide insight into the mechanisms ensuring tissue homeostasis by dynamic control of somatic stem cell activity.


Assuntos
Proteínas Morfogenéticas Ósseas/metabolismo , Proteínas de Drosophila/metabolismo , Núcleosídeo-Difosfato Quinase/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Receptores de Superfície Celular/metabolismo , Transdução de Sinais/fisiologia , Células-Tronco/metabolismo , Animais , Proteínas de Ligação a DNA/metabolismo , Drosophila melanogaster , Feminino , Homeostase/fisiologia , Intestinos/citologia , Modelos Animais , Proteínas do Tecido Nervoso/metabolismo , Regeneração , Fatores de Transcrição/metabolismo
11.
Lett Appl Microbiol ; 69(3): 148-154, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31278768

RESUMO

Consumption of probiotics has an important role in colorectal cancer prevention. In this study, we aimed to explore that the cell wall protein fractions from Lactobacillus paracasei could induce apoptosis on Caco-2 cell line. The cell wall proteins from L. paracasei were fractionated by gel filtration chromatography (F1, F2 and F3) and characterized by polyacrylamide gel electrophoresis (SDS-PAGE). The anticancer properties were evaluated using MTT assay and Annexin V-FITC/PI staining. Administration of L. paracasei increased a significant concentration- and time-dependent anti-proliferative effect on Caco-2 cell line, determined by cell viability assays. However, a dramatic decrease in cell viability of Caco-2 cells was observed at the concentration of 100 µg ml-1 of F1 L. paracasei for 72 h (58% cell viability, P < 0·05) The results showed that F1 L. paracasei could induce apoptosis in Caco-2 cancer cell line by increased in annexin V and propidium iodide staining for 72 h (up to 90·6%, P < 0·001). These results indicated the importance of the anticancer effects of cell wall protein fractions of L. paracasei in human colon carcinoma Caco-2 cell line. Thus, cell wall protein fractions of L. paracasei can be a potential chemotherapeutic agent against Caco-2 cell lines. SIGNIFICANCE AND IMPACT OF THE STUDY: Significance and Impact of the Study: Our findings revealed that the newly identified cell wall protein fractions from probiotic Lactobacillus paracasei inhibit the cell growth of human colon carcinoma cell line (Caco-2), and the results indicated that the cell wall proteins from L. paracasei can be a potential chemotherapeutic agent against Caco-2 cell lines.


Assuntos
Antineoplásicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Parede Celular/metabolismo , Neoplasias do Colo/tratamento farmacológico , Lactobacillus paracasei/metabolismo , Proteínas de Membrana/farmacologia , Apoptose/efeitos dos fármacos , Células CACO-2 , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Intestinos/citologia , Probióticos/farmacologia
12.
Med Sci (Paris) ; 35(6-7): 549-555, 2019.
Artigo em Francês | MEDLINE | ID: mdl-31274085

RESUMO

The study of gut diseases is often limited by the access to human biological tissues and animal models that do not faithfully mimic the human pathologies. In this context, the development of intestinal organoids from human pluripotent stem cells is paving the way of gastrointestinal physiology and digestive disease study. In this review, we recall the embryonic development of the digestive tract and its translation to human pluripotent stem cell differentiation. We also present the different types of intestinal organoids that can be generated, as well as their applications in research.


Assuntos
Intestinos/citologia , Organoides/citologia , Células-Tronco Pluripotentes/citologia , Animais , Diferenciação Celular/fisiologia , Células Cultivadas , Avaliação Pré-Clínica de Medicamentos/métodos , Gastroenteropatias/patologia , Gastroenteropatias/terapia , Trato Gastrointestinal/citologia , Trato Gastrointestinal/crescimento & desenvolvimento , Trato Gastrointestinal/fisiologia , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Células-Tronco Pluripotentes Induzidas/fisiologia , Intestinos/fisiologia , Organoides/fisiologia , Células-Tronco Pluripotentes/fisiologia , Regeneração/fisiologia , Técnicas de Cultura de Tecidos
13.
J Agric Food Chem ; 67(30): 8370-8381, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31271280

RESUMO

Naturally occurring dietary peptides derived from gastrointestinal digestates of common bean milk and yogurt were studied for their bioaccessibility, bioavailability, and anti-inflammatory activity in both Caco-2 mono- and Caco-2/EA.hy926 co-culture cell models. Anti-inflammatory activities of these peptide extracts were found to be strongly associated with cellular uptake by the intestinal epithelial cells. Mechanisms underlying the cellular uptake were studied by examining the role of peptide transporter 1 and calcium sensing reporter. Three peptides, including γ-glutamyl-S-methylcysteine, γ-glutamyl-leucine, and leucine-leucine-valine, were found to be transported across the Caco-2 cell monolayer and detected by liquid chromatography-tandem mass spectrometry. A strong anti-inflammatory effect was observed in the basolateral EA.hy926 cells (co-culture model), as shown in their inhibition of tumor necrosis factor α-induced pro-inflammatory mediators of the nuclear factor κB and mitogen-activated protein kinase signal cascades. The results suggest that these peptides can be absorbed and possibly have systemic inhibition on inflammatory responses in vascular endothelial cells, indicating potential preventive effects on vascular diseases.


Assuntos
Anti-Inflamatórios/metabolismo , Células Endoteliais/metabolismo , Peptídeos/metabolismo , Phaseolus/química , Extratos Vegetais/metabolismo , Iogurte/análise , Transporte Biológico , Células CACO-2 , Técnicas de Cocultura , Células Epiteliais/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Intestinos/citologia , NF-kappa B/genética , NF-kappa B/metabolismo , Transportador 1 de Peptídeos/genética , Transportador 1 de Peptídeos/metabolismo , Peptídeos/química , Phaseolus/metabolismo , Extratos Vegetais/química
14.
Cells ; 8(6)2019 06 15.
Artigo em Inglês | MEDLINE | ID: mdl-31208048

RESUMO

Na-K-ATPase on the basolateral membrane provides the favorable transcellular Na gradient for the proper functioning of Na-dependent nutrient co-transporters on the brush border membrane (BBM) of enterocytes. As cells mature from crypts to villus, Na-K-ATPase activity doubles, to accommodate for the increased BBM Na-dependent nutrient absorption. However, the mechanism of increased Na-K-ATPase activity during the maturation of enterocytes is not known. Therefore, this study aimed to determine the mechanisms involved in the functional transition of Na-K-ATPase during the maturation of crypts to villus cells. Na-K-ATPase activity gradually increased as IEC-18 cells matured in vitro from day 0 (crypts) through day 4 (villus) of post-confluence. mRNA abundance and Western blot studies showed no change in the levels of Na-K-ATPase subunits α1 and ß1 from 0 to 4 days post-confluent cells. However, Na-K-ATPase α1 phosphorylation levels on serine and tyrosine, but not threonine, residues gradually increased. These data indicate that as enterocytes mature from crypt-like to villus-like in culture, the functional activity of Na-K-ATPase increases secondary to altered affinity of the α1 subunit to extracellular K+, in order to accommodate the functional preference of the intestinal cell type. This altered affinity is likely due to increased phosphorylation of the α1 subunit, specifically at serine and tyrosine residues.


Assuntos
Diferenciação Celular , Células Epiteliais/citologia , Células Epiteliais/enzimologia , Intestinos/citologia , ATPase Trocadora de Sódio-Potássio/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Linhagem Celular , Proliferação de Células , Glucose/metabolismo , Cinética , Fosforilação , Subunidades Proteicas/genética , Subunidades Proteicas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Sódio/metabolismo , ATPase Trocadora de Sódio-Potássio/genética
15.
Biol Pharm Bull ; 42(6): 989-995, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31155596

RESUMO

An Intact form of lactoferrin (LF) is known to be absorbed from the small intestine and transported into the blood circulation. We reevaluated the cellular uptake and release of LF using an enterocyte model of human small intestinal cells derived from the Caco-2 cell line. In contrast to a previous report, we observed that intact bovine LF was taken up into seven and 21 d-cultured Caco-2 cells and successfully released back into the culture medium, even though the human intestinal LF receptor, intelectin-1, was not immunochemically detectable. Similar observations were made for human LF and its derivatives (the N-terminal half of LF designated N-lobe and Fc fusions). These observations regarding the uptake and release of intact LF in Caco-2 cells were consistent with in vivo observations. Therefore, we propose that the uptake and release of intact LF by Caco-2 cells should be assessed as a potential in vitro model of in vivo LF absorption in human intestines.


Assuntos
Enterócitos/efeitos dos fármacos , Intestinos/citologia , Lactoferrina/farmacologia , Animais , Células CHO , Células CACO-2 , Cricetulus , Enterócitos/metabolismo , Humanos , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Proteínas Recombinantes/metabolismo
16.
Nat Microbiol ; 4(10): 1737-1749, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31182797

RESUMO

Products derived from bacterial members of the gut microbiota evoke immune signalling pathways of the host that promote immunity and barrier function in the intestine. How immune reactions to enteric viruses support intestinal homeostasis is unknown. We recently demonstrated that infection by murine norovirus (MNV) reverses intestinal abnormalities following depletion of bacteria, indicating that an intestinal animal virus can provide cues to the host that are typically attributed to the microbiota. Here, we elucidate mechanisms by which MNV evokes protective responses from the host. We identify an important role for the viral protein NS1/2 in establishing local replication and a type I interferon (IFN-I) response in the colon. We further show that IFN-I acts on intestinal epithelial cells to increase the proportion of CCR2-dependent macrophages and interleukin (IL)-22-producing innate lymphoid cells, which in turn promote pSTAT3 signalling in intestinal epithelial cells and protection from intestinal injury. In addition, we demonstrate that MNV provides a striking IL-22-dependent protection against early-life lethal infection by Citrobacter rodentium. These findings demonstrate novel ways in which a viral member of the microbiota fortifies the intestinal barrier during chemical injury and infectious challenges.


Assuntos
Microbioma Gastrointestinal/imunologia , Interferon Tipo I/metabolismo , Interleucinas/metabolismo , Intestinos/imunologia , Intestinos/virologia , Animais , Antibacterianos/toxicidade , Proliferação de Células , Citrobacter rodentium/fisiologia , Colo/citologia , Colo/imunologia , Colo/metabolismo , Colo/virologia , Sulfato de Dextrana/toxicidade , Infecções por Enterobacteriaceae/prevenção & controle , Interleucinas/genética , Mucosa Intestinal/citologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/imunologia , Mucosa Intestinal/metabolismo , Intestinos/citologia , Intestinos/efeitos dos fármacos , Linfócitos/citologia , Linfócitos/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Mutação , Norovirus/imunologia , Norovirus/fisiologia , Transdução de Sinais/genética , Organismos Livres de Patógenos Específicos , Proteínas não Estruturais Virais/genética , Replicação Viral
17.
Zhonghua Yu Fang Yi Xue Za Zhi ; 53(6): 638-644, 2019 Jun 06.
Artigo em Chinês | MEDLINE | ID: mdl-31177765

RESUMO

Intestinal microbes play an important role in human health. The development of various clinical diseases, such as obesity, diabetes and cardiovascular disease, is closely related to the imbalance of intestinal microflora. With the development of high-throughput sequencing technology, there has been a breakthrough in the understanding of intestinal microorganism. The interaction between intestinal epithelial cells and intestinal microbes has become one of the hotspots and difficulties of current research. Because of the constraints of ethical review and experimental cost, people are more interested in the development of interaction models between the intestinal microflora and the host cells. In this paper, interaction models between intestinal microflora and host cells, and its working principle and application prospect are reviewed, hoping to provide new techniques and new ideas for studying functions of intestinal microbes.


Assuntos
Diabetes Mellitus , Microbioma Gastrointestinal , Intestinos , Obesidade , Diabetes Mellitus/microbiologia , Células Epiteliais , Humanos , Intestinos/citologia , Intestinos/microbiologia , Obesidade/microbiologia
18.
J Dairy Sci ; 102(8): 6802-6819, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31202650

RESUMO

The process of fermentation contributes to the organoleptic properties, preservation, and nutritional benefits of food. Fermented food may interfere with pathogen infections through a variety of mechanisms, including competitive exclusion or improving intestinal barrier integrity. In this study, the effect of milk fermented with Lactococcus lactis ssp. cremoris JFR1 on Salmonella invasion of intestinal epithelial cell cultures was investigated. Epithelial cells (HT29-MTX, Caco-2, and cocultures of the 2) were treated for 1 h with Lactococcus lactis ssp. cremoris JFR1 fermented milk before infection with Salmonella enterica ssp. enterica Typhimurium. Treatment with fermented milk resulted in increased transepithelial electrical resistance, which remained constant for the duration of infection (up to 3 h), illustrating a protective effect. After gentamicin treatment to remove adhered bacterial cells, enumeration revealed a reduction in numbers of intracellular Salmonella. Quantitative reverse-transcription PCR data indicated a downregulation of Salmonella virulence genes hilA, invA, and sopD after treatment with fermented milk. Fermented milk treatment of epithelial cells also exhibited an immunomodulatory effect reducing the production of proinflammatory IL-8. In contrast, chemically acidified milk (glucono delta-lactone) failed to show the same effect on monolayer integrity, Salmonella Typhimurium invasion, and gene expression as well as immune modulation. Furthermore, an oppA knockout mutant of Salmonella Typhimurium infecting treated epithelial cells did not show suppressed virulence gene expression. Collectively, these results suggest that milk fermented with Lactococcus lactis ssp. cremoris JFR1 is effective in vitro in the reduction of Salmonella invasion into intestinal epithelial cells. A functional OppA permease in Salmonella is required to obtain the antivirulence effect of fermented milk.


Assuntos
Produtos Fermentados do Leite , Fermentação , Intestinos/microbiologia , Lactococcus lactis/metabolismo , Leite/fisiologia , Salmonella typhimurium/fisiologia , Animais , Reatores Biológicos , Células CACO-2 , Células Epiteliais/microbiologia , Expressão Gênica , Humanos , Fatores Imunológicos , Intestinos/citologia , Ácido Láctico/metabolismo , Leite/microbiologia , Salmonella typhimurium/genética , Salmonella typhimurium/patogenicidade , Fatores de Virulência/genética
19.
J Agric Food Chem ; 67(20): 5772-5781, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31046268

RESUMO

This study aimed to purify and identify antioxidant peptides from the low-molecular-weight fraction (SPH-I, MW < 3 kDa) of Alcalase-hydrolyzed soybean ( Glycine max L.) hydrolysate and further evaluate the cytoprotective effects of synthesized peptides against oxidative stress in human intestinal Caco-2 cells. After purification by gel filtration chromatography and reversed-phase HPLC, four major peptides were sequenced by nano-LC-ESI-MS/MS as VVFVDRL (847 Da, SPH-IA), VIYVVDLR (976 Da, SPH-IB), IYVVDLR (877 Da, SPH-IC), and IYVFVR (795 Da, SPH-ID). The antioxidant peptides were synthesized and displayed desirable DPPH radical-scavenging activity (from 16.5 ± 0.5 to 20.3 ± 1.0 µM Trolox equivalent (TE)/µM), ABTS•+ radical-scavenging activity (from 3.42 ± 0.2 to 4.24 ± 0.4 mM TE/µM), ORAC (from 143 ± 2.1 to 171 ± 4.8 µM TE/µM), and FRAP (from 54.7 ± 1.2 to 79.0 ± 0.6 mM Fe2+/µM). Moreover, the synthesized peptides protected Caco-2 cells against H2O2-induced oxidative damage via significantly downregulating intracellular ROS generation and lipid peroxidation ( p < 0.05). Additionally, SPH-IC and SPH-ID statistically upregulated total reduced glutathione synthesis, enhanced activities of catalase and glutathione reductase, and suppressed ROS-mediated inflammatory responses via inhibiting interleukin-8 secretion ( p < 0.05).


Assuntos
Antioxidantes/química , Antioxidantes/farmacologia , Peptídeos/química , Peptídeos/farmacologia , Substâncias Protetoras/química , Substâncias Protetoras/farmacologia , Soja/química , Antioxidantes/isolamento & purificação , Biocatálise , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Glutationa/metabolismo , Humanos , Peróxido de Hidrogênio/toxicidade , Hidrólise , Intestinos/citologia , Intestinos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Peptídeos/isolamento & purificação , Substâncias Protetoras/isolamento & purificação , Subtilisinas/química
20.
Artigo em Inglês | MEDLINE | ID: mdl-31059784

RESUMO

This study evaluated the efficacy of replacing dietary fish oil (FO) with vegetable oils (virgin coconut and corn oil) on enzyme activities (glycolytic, oxidative and lipid metabolites), mRNA expression of lipid metabolic genes and histomorphology of liver and intestine in O. niloticus. O. niloticus (6.07 ±â€¯0.07 g) was fed six experimental diets where fish oil (FO) served as the control diet, and then was supplemented by dietary oils; virgin coconut oil (VCO) {3%FO + 3%VCO; 3FVCO}, and corn oil (CO) {3%FO + 3%CO; 3FCO}, 6%VCO (VCO), 6%CO (CO) and 6%VO {3%VCO + 3%CO; VO}. Growth performances measured indicated fish fed diet 3FCO had higher weight gain (WG) and specific growth rate (SGR). Fish fed diet 3FCO recorded the highest activities in lactate dehydrogenase (LDH), pyruvate kinase (PK), citrate synthase (CS), cytochrome coxidase (COX), malic enzymes (ME) and lipoprotein lipase (LPL) respectively. Stearoyl-CoA desaturase (SCD1) was upregulated in groups fed diets 3FVCO and 3FCO. Also, groups fed diet VCO and CO expressed highly in LPL, whereas, elongase of very long-chain fatty acids (ELOVL-5) was not influenced by the lipid sources. Histological representations in the liver were highly impacted in vegetable diets where lipid accumulation was higher except those fed VCO. However, in the digestive tract from distal to middle and posterior, the same group (VCO) exhibited altered morphological structure as those fed diet 3FCO were similar to FO. The study shows that, corn oil in diets relates positively to growth and enzymatic activities which becomes evident in their depositions in liver and functional intestinal tracts. This study indicates dietary alternatives may cause alterations in lipid metabolic pathways (LPL and SCD1) involved in fatty acid transport. As such, polyunsaturated fatty acid (PUFA) rich diets (CO) based on this study results increases metabolic activities involving especially the production, distribution and consumption of adenosine triphosphate (ATP) in O. niloticus.


Assuntos
Ciclídeos/fisiologia , Gorduras Insaturadas na Dieta/farmacologia , Proteínas de Peixes/genética , Fígado/enzimologia , Músculos/enzimologia , Trifosfato de Adenosina/metabolismo , Ração Animal , Animais , Peso Corporal/efeitos dos fármacos , Ciclídeos/genética , Ciclídeos/crescimento & desenvolvimento , Óleo de Coco/farmacologia , Óleo de Milho/farmacologia , Enzimas/metabolismo , Óleos de Peixe/farmacologia , Proteínas de Peixes/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Intestinos/citologia , Intestinos/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Metabolismo dos Lipídeos/genética , Fígado/citologia , Fígado/efeitos dos fármacos , Músculos/efeitos dos fármacos
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