Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 38.301
Filtrar
1.
Zhongguo Zhong Yao Za Zhi ; 46(3): 670-677, 2021 Feb.
Artículo en Chino | MEDLINE | ID: mdl-33645034

RESUMEN

This study aims to investigate the potential mechanism of curcumin in mediating interleukin-6(IL-6)/signal transducer and activator of transcription 3(STAT3) signaling pathway to repair intestinal mucosal injury induced by 5-fluorouracil(5-FU) chemotherapy for colon cancer. SD rats were intraperitoneally injected with 60 mg·kg~(-1)·d~(-1) 5-FU for 4 days to establish a model of intestinal mucosal injury. Then the rats were randomly divided into model group(equal volume of normal saline), curcumin low, medium and high dose groups(50, 100, 200 mg·kg~(-1)), and normal SD rats were used as control group(equal volume of normal saline). Each group received gavage administration for 4 consecutive days, and the changes of body weight and feces were recorded every day. After administration, blood was collected from the heart, and jejunum tissues were collected. The levels of serum interleukin-1ß(IL-1ß) and tumor necrosis factor-α(TNF-α) were detected by ELISA, and at the same time, the concentration of Evans blue(EB) in jejunum was measured. Hematoxylin-eosin(HE) staining was used to observe the pathological state of jejunum, and the length of jejunum villi and the depth of crypt were measured. The positive expression levels of claudin, occludin and ZO-1 were detected by immunohistochemistry. Western blot was used to detect the protein expression of IL-6, p-STAT3, E-cadherin, vimentin and N-cadherin in jejunum tissues. The results showed that, curcumin significantly increased body weight and fecal weight(P<0.05 or P<0.01), decreased fecal score, EB concentration, IL-1ß and TNF-α levels(P<0.05 or P<0.01) in rats. In addition, curcumin maintained the integrity of mucosal surface and villi structure of jejunum to a large extent, and reduced pathological changes in a dose-dependent manner. Meanwhile, curcumin could increase the positive expression of occludin, claudin and ZO-1(P<0.05 or P<0.01), repair intestinal barrier function, downregulate the protein expression of IL-6, p-STAT3, vimentin and N-cadherin in jejunum tissues(P<0.05 or P<0.01), and upregulate the protein expression of E-cadherin(P<0.05). Therefore, curcumin could repair the intestinal mucosal injury induced by 5-FU chemotherapy for colon cancer, and the mechanism may be related to the inhibition of IL-6/STAT3 signal and the inhibition of epithelial-mesenchymal transition(EMT) process.


Asunto(s)
Neoplasias del Colon , Curcumina , Animales , Neoplasias del Colon/tratamiento farmacológico , Fluorouracilo/toxicidad , Interleucina-6/genética , Mucosa Intestinal/metabolismo , Ratas , Ratas Sprague-Dawley , Factor de Transcripción STAT3/genética , Factor de Transcripción STAT3/metabolismo , Transducción de Señal
2.
Methods Mol Biol ; 2291: 273-283, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33704758

RESUMEN

The environment in the human intestine is low in oxygen. This affects virulence gene expression of enteropathogens including Shiga toxin-producing E. coli (STEC) and enables mucosal colonization by oxygen-sensitive commensal bacteria. To simulate the oxygen-restricted milieu at the intestinal epithelium, we have developed a vertical diffusion chamber model (VDC) which allows infection of polarized human intestinal epithelia under microaerobic conditions. In this chapter, we present a detailed protocol for performing STEC infections in the VDC system and subsequent analysis of STEC pathogenesis.


Asunto(s)
Mucosa Intestinal , Modelos Biológicos , Oxígeno/metabolismo , Escherichia coli Shiga-Toxigénica/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología
3.
Methods Mol Biol ; 2291: 285-296, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33704759

RESUMEN

Human intestinal organoid cultures established from crypt-derived stem cells truly revolutionized our approach to study intestinal epithelial physiology and pathologies as they can be propagated indefinitely and preserve the genetic signature of the donor and the gut segment specificity in culture. Here we describe human stem cell-derived colonoid monolayers as a reliable and reproducible model to study Shiga toxin-producing Escherichia coli (STEC) infection and STEC-caused pathologies of the whole colonic epithelium comprising a mixture of colonocytes, goblet, enteroendocrine, and other rare cells present in human colonic epithelial tissue.


Asunto(s)
Colon , Células Epiteliales , Infecciones por Escherichia coli/metabolismo , Interacciones Huésped-Patógeno , Mucosa Intestinal , Modelos Biológicos , Escherichia coli Shiga-Toxigénica/fisiología , Colon/metabolismo , Colon/microbiología , Células Epiteliales/metabolismo , Células Epiteliales/microbiología , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología
4.
Methods Mol Biol ; 2291: 381-397, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33704765

RESUMEN

Shiga toxin-producing E. coli (STEC) is a common foodborne pathogen in developed countries. STEC generates "attaching and effacing" (AE) lesions on colonic epithelium, characterized by effacement of microvilli and the formation of actin "pedestals" beneath intimately attached bacteria. In addition, STEC are lysogenized with a phage that, upon induction, can produce potent Shiga toxins (Stx), potentially leading to both hemorrhagic colitis and hemolytic uremic syndrome. Investigation of the pathogenesis of this disease has been challenging because STEC does not readily colonize conventional mice.Citrobacter rodentium (CR) is a related mouse pathogen that also generates AE lesions. Whereas CR does not produce Stx, a murine model for STEC utilizes CR lysogenized with an E. coli-derived Stx phage, generating CR(Φstx), which both colonizes conventional mice and readily gives rise to systemic disease. We present here key methods for the use of CR(Φstx) infection as a highly predictable murine model for infection and disease by STEC. Importantly, we detail CR(Φstx) inoculation by feeding, determination of pathogen colonization, production of phage and toxin, and assessment of intestinal and renal pathology. These methods provide a framework for studying STEC-mediated systemic disease that may aid in the development of efficacious therapeutics.


Asunto(s)
Bacteriófagos , Citrobacter rodentium , Colitis , Hemorragia Gastrointestinal , Síndrome Hemolítico-Urémico , Mucosa Intestinal , Lisogenia , Toxinas Shiga , Escherichia coli Shiga-Toxigénica , Animales , Bacteriófagos/genética , Bacteriófagos/metabolismo , Citrobacter rodentium/genética , Citrobacter rodentium/metabolismo , Citrobacter rodentium/patogenicidad , Citrobacter rodentium/virología , Colitis/genética , Colitis/metabolismo , Colitis/microbiología , Modelos Animales de Enfermedad , Hemorragia Gastrointestinal/genética , Hemorragia Gastrointestinal/metabolismo , Hemorragia Gastrointestinal/microbiología , Síndrome Hemolítico-Urémico/genética , Síndrome Hemolítico-Urémico/metabolismo , Síndrome Hemolítico-Urémico/microbiología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiología , Ratones , Toxinas Shiga/biosíntesis , Toxinas Shiga/genética
5.
Nat Commun ; 12(1): 1067, 2021 02 16.
Artículo en Inglés | MEDLINE | ID: mdl-33594081

RESUMEN

Increases in adhesive and invasive commensal bacteria, such as Escherichia coli, and subsequent disruption of the epithelial barrier is implicated in the pathogenesis of inflammatory bowel disease (IBD). However, the protective systems against such barrier disruption are not fully understood. Here, we show that secretion of luminal glycoprotein 2 (GP2) from pancreatic acinar cells is induced in a TNF-dependent manner in mice with chemically induced colitis. Fecal GP2 concentration is also increased in Crohn's diease patients. Furthermore, pancreas-specific GP2-deficient colitis mice have more severe intestinal inflammation and a larger mucosal E. coli population than do intact mice, indicating that digestive-tract GP2 binds commensal E. coli, preventing epithelial attachment and penetration. Thus, the pancreas-intestinal barrier axis and pancreatic GP2 are important as a first line of defense against adhesive and invasive commensal bacteria during intestinal inflammation.


Asunto(s)
Inflamación/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Glicoproteínas de Membrana/metabolismo , Células Acinares/metabolismo , Células Acinares/patología , Animales , Colitis/metabolismo , Colitis/patología , Citocinas/metabolismo , Sulfato de Dextran , Escherichia coli/efectos de los fármacos , Escherichia coli/fisiología , Heces , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Inmunoglobulina A/metabolismo , Mucosa Intestinal/microbiología , Ratones Endogámicos C57BL , Páncreas/patología , Proteínas Recombinantes/farmacología , Factores de Transcripción/metabolismo , Regulación hacia Arriba/genética
6.
Nat Commun ; 12(1): 836, 2021 02 05.
Artículo en Inglés | MEDLINE | ID: mdl-33547321

RESUMEN

Dynamic regulation of intestinal cell differentiation is crucial for both homeostasis and the response to injury or inflammation. Sprouty2, an intracellular signaling regulator, controls pathways including PI3K and MAPKs that are implicated in differentiation and are dysregulated in inflammatory bowel disease. Here, we ask whether Sprouty2 controls secretory cell differentiation and the response to colitis. We report that colonic epithelial Sprouty2 deletion leads to expanded tuft and goblet cell populations. Sprouty2 loss induces PI3K/Akt signaling, leading to GSK3ß inhibition and epithelial interleukin (IL)-33 expression. In vivo, this results in increased stromal IL-13+ cells. IL-13 in turn induces tuft and goblet cell expansion in vitro and in vivo. Sprouty2 is downregulated by acute inflammation; this appears to be a protective response, as VillinCre;Sprouty2F/F mice are resistant to DSS colitis. In contrast, Sprouty2 is elevated in chronic colitis and in colons of inflammatory bowel disease patients, suggesting that this protective epithelial-stromal signaling mechanism is lost in disease.


Asunto(s)
Colitis/genética , Glucógeno Sintasa Quinasa 3 beta/genética , Homeostasis/genética , Interleucina-33/genética , Proteínas de la Membrana/genética , Proteínas Serina-Treonina Quinasas/genética , Animales , Recuento de Células , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Niño , Colitis/inducido químicamente , Colitis/metabolismo , Colitis/patología , Colon/efectos de los fármacos , Colon/metabolismo , Colon/patología , Femenino , Regulación de la Expresión Génica , Glucógeno Sintasa Quinasa 3 beta/metabolismo , Células Caliciformes/efectos de los fármacos , Células Caliciformes/metabolismo , Células Caliciformes/patología , Células HT29 , Homeostasis/efectos de los fármacos , Humanos , Interleucina-33/metabolismo , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Masculino , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Proteínas de Microfilamentos/genética , Proteínas de Microfilamentos/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Transducción de Señal , Dodecil Sulfato de Sodio/administración & dosificación
7.
Medicine (Baltimore) ; 100(4): e24078, 2021 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-33530201

RESUMEN

BACKGROUND: Ulcerative colitis (UC) is a chronic nonspecific intestinal inflammatory disease with unclear etiology occurring in the colonic mucosa. Its clinical manifestations are characterized by recurrent abdominal pain, diarrhea, mucous pus, and blood stool. The severity of the disease varies, and itis characterized by a high recurrence rate. Because of its long course of disease, easy to relapse, protracted and difficult to recover, seriously affect the quality of life, increase the economic burden of patients and society, and even the risk of developing cancer, it has become one of the hot issues of general concern in the medical field. Heat-sensitive moxibustion therapy has shown strong advantages in the treatment of UC, and the curative effect is accurate. therefore, this paper will carry out a systematic evaluation and meta analysis of the efficacy and safety of heat-sensitive moxibustion in the treatment of UC. METHODS: We will be searching 8 electronic databases, including PubMed, Embase, Web of Science, Cochrane Library, the China National Knowledge Infrastructure, Chinese Science and Technology Periodical Database, Wanfang Database, and Chinese Biomedical Literature Database. We will search above electronic databases from the beginning to December 2020, without any language restriction. Clinical efficacy, including total effective rate or cure rate, clinical symptom integral (abdominal pain, diarrhea, purulent stool), and recurrence rate will be accepted as the primary outcomes. The changes of cytokine Hs-CRP, IL-6, TNF-αlevels in serum, and improvement of colorectal mucosa will be used as secondary outcomes. RevMan 5.3 software will be used for statistical analysis. The result about the curative effect and safety of heat-sensitive moxibustion for UC will be presented as risk ratio for dichotomous data and mean differences with a 95% confidence interval for continuous data. RESULTS: When this research program is completed, the relevant results can be obtained. CONCLUSIONS: The results of this study will provide reliable evidence for the efficacy and safety of heat-sensitive moxibustion in the treatment of UC. INPLASY REGISTRATION NUMBER: INPLASY20201101034.


Asunto(s)
Colitis Ulcerosa/terapia , Moxibustión/métodos , Citocinas/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Moxibustión/efectos adversos , Calidad de Vida , Ensayos Clínicos Controlados Aleatorios como Asunto , Recurrencia , Proyectos de Investigación , Índice de Severidad de la Enfermedad , Temperatura
8.
Int J Mol Sci ; 22(2)2021 Jan 08.
Artículo en Inglés | MEDLINE | ID: mdl-33435615

RESUMEN

Celiac disease is an autoimmune disorder characterized by a heightened immune response to gluten proteins in the diet, leading to gastrointestinal symptoms and mucosal damage localized to the small intestine. Despite its prevalence, the only treatment currently available for celiac disease is complete avoidance of gluten proteins in the diet. Ongoing clinical trials have focused on targeting the immune response or gluten proteins through methods such as immunosuppression, enhanced protein degradation and protein sequestration. Recent studies suggest that polyphenols may elicit protective effects within the celiac disease milieu by disrupting the enzymatic hydrolysis of gluten proteins, sequestering gluten proteins from recognition by critical receptors in pathogenesis and exerting anti-inflammatory effects on the system as a whole. This review highlights mechanisms by which polyphenols can protect against celiac disease, takes a critical look at recent works and outlines future applications for this potential treatment method.


Asunto(s)
Enfermedades Autoinmunes/inmunología , Enfermedad Celíaca/inmunología , Gliadina/inmunología , Polifenoles/inmunología , Enfermedades Autoinmunes/metabolismo , Enfermedades Autoinmunes/prevención & control , Enfermedad Celíaca/metabolismo , Enfermedad Celíaca/prevención & control , Gliadina/metabolismo , Glútenes/inmunología , Glútenes/metabolismo , Humanos , Inmunosupresión/métodos , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Intestino Delgado/efectos de los fármacos , Intestino Delgado/inmunología , Intestino Delgado/metabolismo , Polifenoles/metabolismo , Polifenoles/uso terapéutico , Estudios Prospectivos
9.
Carbohydr Polym ; 256: 117504, 2021 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-33483027

RESUMEN

Oral administration of insulin (INS) would represent a revolution in the treatment of diabetes, considering that this route mimics the physiological dynamics of endogenous INS. Nano- and microencapsulation exploiting the advantageous polysaccharides properties has been considered an important technological strategy to protect INS against harsh conditions of gastrointestinal tract, in the same time that improve the permeability via transcellular and/or paracellular pathways, safety and in some cases even selectivity for targeting delivery of INS. In fact, some polysaccharides also give to the systems functional properties such as pH-responsiveness, mucoadhesiveness under specific physiological conditions and increased intestinal permeability. In general, all polysaccharides can be functionalized with specific molecules becoming more selective to the cells to which INS is delivered. The present review highlights the advances in the past 10 years on micro- and nanoencapsulation of INS exploiting the unique natural properties of polysaccharides, including chitosan, starch, alginate, pectin, and dextran, among others.


Asunto(s)
Diabetes Mellitus Experimental/tratamiento farmacológico , Diabetes Mellitus/tratamiento farmacológico , Sistemas de Liberación de Medicamentos/métodos , Hipoglucemiantes/farmacocinética , Insulina/farmacocinética , Nanopartículas/química , Administración Oral , Alginatos/química , Animales , Quitosano/química , Dextranos/química , Diabetes Mellitus/metabolismo , Diabetes Mellitus/patología , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/patología , Composición de Medicamentos/métodos , Liberación de Fármacos , Humanos , Hipoglucemiantes/metabolismo , Insulina/metabolismo , Mucosa Intestinal/metabolismo , Nanopartículas/administración & dosificación , Pectinas/química , Permeabilidad , Almidón/química
10.
Science ; 371(6524)2021 01 01.
Artículo en Inglés | MEDLINE | ID: mdl-33384352

RESUMEN

Tissue homeostasis is perturbed in a diversity of inflammatory pathologies. These changes can elicit endoplasmic reticulum (ER) stress, protein misfolding, and cell death. ER stress triggers the unfolded protein response (UPR), which can promote recovery of ER proteostasis and cell survival or trigger programmed cell death. Here, we leveraged single-cell RNA sequencing to define dynamic transcriptional states associated with the adaptive versus terminal UPR in the mouse intestinal epithelium. We integrated these transcriptional programs with genome-scale CRISPR screening to dissect the UPR pathway functionally. We identified QRICH1 as a key effector of the PERK-eIF2α axis of the UPR. QRICH1 controlled a transcriptional program associated with translation and secretory networks that were specifically up-regulated in inflammatory pathologies. Thus, QRICH1 dictates cell fate in response to pathological ER stress.


Asunto(s)
Proteínas de Unión al ADN/metabolismo , Estrés del Retículo Endoplásmico/genética , Regulación de la Expresión Génica , Inflamación/metabolismo , Proteostasis/genética , Factores de Transcripción/metabolismo , Respuesta de Proteína Desplegada/genética , Animales , Apoptosis , Células Cultivadas , Proteínas de Unión al ADN/genética , Factor 2 Eucariótico de Iniciación/metabolismo , Humanos , Inflamación/genética , Inflamación/patología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Ratones , Organoides , RNA-Seq , Análisis de la Célula Individual , Factores de Transcripción/genética , Transcripción Genética , eIF-2 Quinasa/metabolismo
11.
Nat Genet ; 53(1): 16-26, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33414552

RESUMEN

Oncogenic KRAS mutations and inactivation of the APC tumor suppressor co-occur in colorectal cancer (CRC). Despite efforts to target mutant KRAS directly, most therapeutic approaches focus on downstream pathways, albeit with limited efficacy. Moreover, mutant KRAS alters the basal metabolism of cancer cells, increasing glutamine utilization to support proliferation. We show that concomitant mutation of Apc and Kras in the mouse intestinal epithelium profoundly rewires metabolism, increasing glutamine consumption. Furthermore, SLC7A5, a glutamine antiporter, is critical for colorectal tumorigenesis in models of both early- and late-stage metastatic disease. Mechanistically, SLC7A5 maintains intracellular amino acid levels following KRAS activation through transcriptional and metabolic reprogramming. This supports the increased demand for bulk protein synthesis that underpins the enhanced proliferation of KRAS-mutant cells. Moreover, targeting protein synthesis, via inhibition of the mTORC1 regulator, together with Slc7a5 deletion abrogates the growth of established Kras-mutant tumors. Together, these data suggest SLC7A5 as an attractive target for therapy-resistant KRAS-mutant CRC.


Asunto(s)
Neoplasias Colorrectales/genética , Transportador de Aminoácidos Neutros Grandes 1/metabolismo , Mutación/genética , Proteínas Proto-Oncogénicas p21(ras)/genética , Regiones no Traducidas 5'/genética , Sistema de Transporte de Aminoácidos ASC/metabolismo , Animales , Carcinogénesis/patología , Proliferación Celular , Neoplasias Colorrectales/patología , Regulación Neoplásica de la Expresión Génica , Glutamina/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Estimación de Kaplan-Meier , Diana Mecanicista del Complejo 1 de la Rapamicina/metabolismo , Ratones Endogámicos C57BL , Antígenos de Histocompatibilidad Menor/metabolismo , Metástasis de la Neoplasia , Oncogenes , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismo
12.
Virology ; 556: 1-8, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33515858

RESUMEN

Porcine deltacoronavirus (PDCoV) is one of the emerged coronaviruses posing a significant threat to the swine industry. Previous work showed the presence of a viral accessory protein NS6 in PDCoV-infected cells. In this study, we detected the expression of the NS6 protein in small intestinal tissues of PDCoV-infected piglets. In addition, SDS-PAGE and Western blot analysis of sucrose gradient-purified virions showed the presence of a 13-kDa NS6 protein. Further evidences of the presence of NS6 in the PDCoV virions were obtained by immunogold staining of purified virions with anti-NS6 antiserum, and by immunoprecipitation of NS6 from purified virions. Finally, the anti-NS6 antibody was not able to neutralize PDCoV in cultured cells. These data establish for the first time that the accessory protein NS6 is expressed during infection in vivo and incorporated into PDCoV virions.


Asunto(s)
Infecciones por Coronavirus/veterinaria , Enfermedades de los Porcinos/virología , Proteínas no Estructurales Virales/metabolismo , Virión/metabolismo , Animales , Anticuerpos Antivirales/inmunología , Línea Celular , Infecciones por Coronavirus/metabolismo , Infecciones por Coronavirus/virología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/virología , Ratones , Conejos , Porcinos , Enfermedades de los Porcinos/metabolismo , Proteínas no Estructurales Virales/inmunología
13.
Life Sci ; 267: 118974, 2021 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-33385407

RESUMEN

AIM: We aimed to determine whether the sodium/glucose cotransporter family member SGLT3, a proposed glucose sensor, is expressed in the intestine and/or kidney, and if its expression is altered in mouse models of obesity and in humans before and after weight-loss surgery. MAIN METHODS: We used in-situ hybridization and quantitative PCR to determine whether the Sglt3 isoforms 3a and 3b were expressed in the intestine and kidney of C57, leptin-deficient ob/ob, and diabetic BTBR ob/ob mice. Western blotting and immunohistochemistry were also used to assess SGLT3 protein levels in jejunal biopsies from obese patients before and after weight-loss Roux-en-Y gastric bypass surgery (RYGB), and in lean healthy controls. KEY FINDINGS: Sglt3a/3b mRNA was detected in the small intestine (duodenum, jejunum and ileum), but not in the large intestine or kidneys of mice. Both isoforms were detected in epithelial cells (confirmed using intestinal organoids). Expression of Sglt3a/3b mRNA in duodenum and jejunum was significantly lower in ob/ob and BTBR ob/ob mice than in normal-weight littermates. Jejunal SGLT3 protein levels in aged obese patients before RYGB were lower than in lean individuals, but substantially upregulated 6 months post-RYGB. SIGNIFICANCE: Our study shows that Sglt3a/3b is expressed primarily in epithelial cells of the small intestine in mice. Furthermore, we observed an association between intestinal mRNA Sglt3a/3b expression and obesity in mice, and between jejunal SGLT3 protein levels and obesity in humans. Further studies are required to determine the possible role of SGLT3 in obesity.


Asunto(s)
Obesidad/metabolismo , Proteínas de Transporte de Sodio-Glucosa/genética , Adulto , Animales , Modelos Animales de Enfermedad , Regulación hacia Abajo , Femenino , Derivación Gástrica , Expresión Génica , Humanos , Insulina/metabolismo , Resistencia a la Insulina , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Yeyuno/metabolismo , Leptina/deficiencia , Leptina/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Obesidad/genética , Isoformas de Proteínas , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteínas de Transporte de Sodio-Glucosa/biosíntesis , Proteínas de Transporte de Sodio-Glucosa/metabolismo , Transcriptoma , Pérdida de Peso
14.
Medicina (Kaunas) ; 57(1)2021 Jan 09.
Artículo en Inglés | MEDLINE | ID: mdl-33435405

RESUMEN

Nowadays, humanity faces one of the most serious health crises, the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. The severity of coronavirus disease 2019 (COVID-19) pandemic is related to the high rate of interhuman transmission of the virus, variability of clinical presentation, and the absence of specific therapeutic methods. COVID-19 can manifest with non-specific symptoms and signs, especially among the elderly. In some cases, the clinical manifestations of hyponatremia may be the first to appear. The pathophysiological mechanisms of hyponatremia among patients with COVID-19 are diverse, including syndrome of inappropriate antidiuretic hormone secretion (SIADH), digestive loss of sodium ions, reduced sodium ion intake or use of diuretic therapy. Hyponatremia may also be considered a negative prognostic factor in patients diagnosed with COVID-19. We need further studies to evaluate the etiology and therapeutic management of hyponatremia in patients with COVID-19.


Asunto(s)
/metabolismo , Hiponatremia/metabolismo , Síndrome de Secreción Inadecuada de ADH/metabolismo , /complicaciones , Diuréticos/efectos adversos , Fluidoterapia/métodos , Humanos , Hiponatremia/epidemiología , Hiponatremia/etiología , Hiponatremia/terapia , Síndrome de Secreción Inadecuada de ADH/etiología , Incidencia , Interleucina-6/metabolismo , Mucosa Intestinal/metabolismo , Pronóstico , Solución Salina Hipertónica/uso terapéutico , Sodio en la Dieta
15.
Mol Immunol ; 131: 23-32, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33465592

RESUMEN

Necrotizing enterocolitis (NEC) is a devastating neonatal gastrointestinal emergency. Fucosylated glycans on intestinal epithelial cells (IECs) play a central role in the maintenance of intestinal homeostasis. Nevertheless, its association with necrotizing enterocolitis is not clear. We examined paraffin-embedded intestinal specimens from participants and found that the NEC patients showed lower intestinal epithelial fucosylation levels than the control patients. In the mouse model of NEC, the percentage of fucosylated epithelial cells (F-ECs) and ILC3s was decreased. Also, the expression levels of IL-22 and Fut2 were reduced. Moreover, the critical role of epithelial fucosylation in NEC was further confirmed by administering the anti-IL-22 antibody, which caused an increase in histological damage, body weight loss, intestinal permeability and proinflammatory cytokine release correlated with a reduction of F-ECs. Overall, intestinal fucosylation deficiency led to increased susceptibility and severity of NEC. Further studies are needed to determine whether modification of intestinal fucosylation affects the development of NEC.


Asunto(s)
Enterocolitis Necrotizante/patología , Células Epiteliales/patología , Mucosa Intestinal/patología , Intestinos/patología , Animales , Preescolar , Citocinas/metabolismo , Modelos Animales de Enfermedad , Enterocolitis Necrotizante/metabolismo , Células Epiteliales/metabolismo , Femenino , Fucosiltransferasas/metabolismo , Humanos , Inflamación/metabolismo , Inflamación/patología , Interleucinas/metabolismo , Mucosa Intestinal/metabolismo , Masculino , Ratones
16.
Vet Res ; 52(1): 2, 2021 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-33397461

RESUMEN

Porcine epidemic diarrhea (PED) is a coronavirus disease characterized by the rapid spread of severe diarrhea among pigs. PED virus (PEDV) infects and replicates mainly in the epithelial cells of the duodenum, jejunum, ileum and colon. Serum or mucosal IgA antibody levels have been used to predict both vaccine efficacy and the level of protective immunity to enteric infectious diseases in individuals or herds. Details of the B-cell immune response upon PEDV infection, such as the systemic and mucosal PEDV IgA antibody response, the distribution of IgA antibody-secreting cells (ASCs), and their role in virus clearance are not yet clear. In this experimental infection study, we observed similar fluctuations in PEDV IgA antibody levels in serum and intestinal contents of the upper and lower jejunum and ileum, but not fecal samples, over the 4-week experimental course. ASCs that actively secrete PEDV IgA antibody without in vitro stimulation were distributed mainly in the upper jejunum, whereas memory B cells that showed enhanced PEDV IgA antibody production upon in vitro stimulation were observed in mesenteric lymph nodes and the ileum. Our findings will contribute to the development of effective vaccines and diagnostic methods for PEDV.


Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Coronavirus/veterinaria , Virus de la Diarrea Epidémica Porcina , Enfermedades de los Porcinos/virología , Animales , Chlorocebus aethiops , Infecciones por Coronavirus/sangre , Infecciones por Coronavirus/inmunología , Infecciones por Coronavirus/virología , Heces/química , Heces/virología , Inmunoglobulina A/sangre , Inmunoglobulina A/química , Inmunoglobulina A/metabolismo , Inmunoglobulina G/sangre , Mucosa Intestinal/metabolismo , ARN Viral , Porcinos , Enfermedades de los Porcinos/sangre , Enfermedades de los Porcinos/inmunología , Células Vero
17.
Nat Commun ; 12(1): 134, 2021 01 08.
Artículo en Inglés | MEDLINE | ID: mdl-33420022

RESUMEN

Understanding the factors that contribute to efficient SARS-CoV-2 infection of human cells may provide insights on SARS-CoV-2 transmissibility and pathogenesis, and reveal targets of intervention. Here, we analyze host and viral determinants essential for efficient SARS-CoV-2 infection in both human lung epithelial cells and ex vivo human lung tissues. We identify heparan sulfate as an important attachment factor for SARS-CoV-2 infection. Next, we show that sialic acids present on ACE2 prevent efficient spike/ACE2-interaction. While SARS-CoV infection is substantially limited by the sialic acid-mediated restriction in both human lung epithelial cells and ex vivo human lung tissues, infection by SARS-CoV-2 is limited to a lesser extent. We further demonstrate that the furin-like cleavage site in SARS-CoV-2 spike is required for efficient virus replication in human lung but not intestinal tissues. These findings provide insights on the efficient SARS-CoV-2 infection of human lungs.


Asunto(s)
/metabolismo , /transmisión , Ácidos Siálicos/metabolismo , Glicoproteína de la Espiga del Coronavirus/metabolismo , Acoplamiento Viral , Animales , Células CACO-2 , Línea Celular Tumoral , Chlorocebus aethiops , Cricetinae , Furina/metabolismo , Células HEK293 , Heparitina Sulfato/metabolismo , Humanos , Mucosa Intestinal/metabolismo , Intestinos/virología , Pulmón/patología , Pulmón/virología , Síndrome Respiratorio Agudo Grave/patología , Células Vero , Internalización del Virus , Replicación Viral/fisiología
18.
Int J Nanomedicine ; 16: 345-357, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33488076

RESUMEN

Background: Our previous study found that deletion of Sorting nexin 10 (SNX10) can protect against colonic inflammation and pathological damage induced by dextran sulfate sodium (DSS). This inspired us that modulation of SNX10 expression in colonic epithelial cells might represent a promising therapeutic strategy for inflammatory bowel disease (IBD). Methods: Effective delivery of siRNA/shRNA to silence genes is a highly sought-after means in the treatment of multiple diseases. Here, we encapsulated SNX10-shRNA plasmids (SRP) with polylactide-polyglycolide (PLGA) to make oral nanoparticles (NPs), and then applied them to acute and chronic IBD mice model, respectively. The characteristics of the nanoparticles were assayed and the effects of SRP-NPs on mouse IBD were evaluated. Results: High-efficiency SNX10-shRNA plasmids were successfully constructed and coated with PLGA to obtain nanoparticles, with a particle size of 275.2 ± 11.4mm, uniform PDI distribution, entrapment efficiency of 87.6 ± 2.5%, and drug loading of 13.11 ± 1.38%, displayed dominant efficiency of SNX10 RNA interference in the colon. In both acute and chronic IBD models, SRP-NPs could effectively reduce the loss of mice body weight, relieve the intestinal mucosal damage and inflammatory infiltration, inhibit the expression of inflammatory cytokines IL-1ß, IL-23, TNF-α, and down-regulate the expression of toll-like receptors (TLRs) 2 and 4. Conclusion: Oral nanoparticles of SNX10-shRNA plasmid displayed dominant efficiency of SNX10 RNA interference in the colon and ameliorate mouse colitis via TLR signaling pathway. SNX10 is a new target for IBD treatment and nanoparticles of SNX10-shRNA plasmid might be a promising treatment option for IBD.


Asunto(s)
Colitis/terapia , Portadores de Fármacos/química , Terapia Genética/métodos , Nanopartículas/química , Plásmidos/genética , ARN Interferente Pequeño/administración & dosificación , ARN Interferente Pequeño/genética , Administración Oral , Animales , Colitis/inducido químicamente , Colitis/genética , Colitis/metabolismo , Citocinas/metabolismo , Modelos Animales de Enfermedad , Mucosa Intestinal/metabolismo , Ratones , Interferencia de ARN , ARN Interferente Pequeño/química , Transducción de Señal/genética , Factor de Necrosis Tumoral alfa/metabolismo
19.
Nat Cell Biol ; 23(1): 23-31, 2021 01.
Artículo en Inglés | MEDLINE | ID: mdl-33398177

RESUMEN

A detailed understanding of intestinal stem cell (ISC) self-renewal and differentiation is required to treat chronic intestinal diseases. However, the different models of ISC lineage hierarchy1-6 and segregation7-12 are subject to debate. Here, we have discovered non-canonical Wnt/planar cell polarity (PCP)-activated ISCs that are primed towards the enteroendocrine or Paneth cell lineage. Strikingly, integration of time-resolved lineage labelling with single-cell gene expression analysis revealed that both lineages are directly recruited from ISCs via unipotent transition states, challenging the existence of formerly predicted bi- or multipotent secretory progenitors7-12. Transitory cells that mature into Paneth cells are quiescent and express both stem cell and secretory lineage genes, indicating that these cells are the previously described Lgr5+ label-retaining cells7. Finally, Wnt/PCP-activated Lgr5+ ISCs are molecularly indistinguishable from Wnt/ß-catenin-activated Lgr5+ ISCs, suggesting that lineage priming and cell-cycle exit is triggered at the post-transcriptional level by polarity cues and a switch from canonical to non-canonical Wnt/PCP signalling. Taken together, we redefine the mechanisms underlying ISC lineage hierarchy and identify the Wnt/PCP pathway as a new niche signal preceding lateral inhibition in ISC lineage priming and segregation.


Asunto(s)
Linaje de la Célula , Polaridad Celular , Células Enteroendocrinas/citología , Mucosa Intestinal/citología , Células de Paneth/citología , Células Madre/citología , Proteínas Wnt/metabolismo , Animales , Autorrenovación de las Células , Células Enteroendocrinas/metabolismo , Femenino , Perfilación de la Expresión Génica , Mucosa Intestinal/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Células de Paneth/metabolismo , Receptores Acoplados a Proteínas G/fisiología , Análisis de la Célula Individual , Células Madre/metabolismo , beta Catenina/metabolismo
20.
Am J Physiol Gastrointest Liver Physiol ; 320(3): G396-G410, 2021 03 01.
Artículo en Inglés | MEDLINE | ID: mdl-33355506

RESUMEN

Poor translatability of animal disease models has hampered the development of new inflammatory bowel disorder (IBD) therapeutics. We describe a preclinical, ex vivo system using freshly obtained and well-characterized human colorectal tissue from patients with ulcerative colitis (UC) and healthy control (HC) participants to test potential therapeutics for efficacy and target engagement, using the JAK/STAT inhibitor tofacitinib (TOFA) as a model therapeutic. Colorectal biopsies from HC participants and patients with UC were cultured and stimulated with multiple mitogens ± TOFA. Soluble biomarkers were detected using a 29-analyte multiplex ELISA. Target engagement in CD3+CD4+ and CD3+CD8+ T-cells was determined by flow cytometry in peripheral blood mononuclear cells (PBMCs) and isolated mucosal mononuclear cells (MMCs) following the activation of STAT1/3 phosphorylation. Data were analyzed using linear mixed-effects modeling, t test, and analysis of variance. Biomarker selection was performed using penalized and Bayesian logistic regression modeling, with results visualized using uniform manifold approximation and projection. Under baseline conditions, 27 of 29 biomarkers from patients with UC were increased versus HC participants. Explant stimulation increased biomarker release magnitude, expanding the dynamic range for efficacy and target engagement studies. Logistic regression analyses identified the most representative UC baseline and stimulated biomarkers. TOFA inhibited biomarkers dependent on JAK/STAT signaling. STAT1/3 phosphorylation in T-cells revealed compartmental differences between PBMCs and MMCs. Immunogen stimulation increases biomarker release in similar patterns for HC participants and patients with UC, while enhancing the dynamic range for pharmacological effects. This work demonstrates the power of ex vivo human colorectal tissue as preclinical tools for evaluating target engagement and downstream effects of new IBD therapeutic agents.NEW & NOTEWORTHY Using colorectal biopsy material from healthy volunteers and patients with clinically defined IBD supports translational research by informing the evaluation of therapeutic efficacy and target engagement for the development of new therapeutic entities. Combining experimental readouts from intact and dissociated tissue enhances our understanding of the tissue-resident immune system that contribute to disease pathology. Bayesian logistic regression modeling is an effective tool for predicting ex vivo explant biomarker release patterns.


Asunto(s)
Colitis Ulcerosa/metabolismo , Citocinas/metabolismo , Mucosa Intestinal/efectos de los fármacos , Piperidinas/farmacología , Inhibidores de Proteínas Quinasas/farmacología , Pirimidinas/farmacología , Linfocitos T/efectos de los fármacos , Teorema de Bayes , Biomarcadores , Colitis Ulcerosa/patología , Citocinas/antagonistas & inhibidores , Citocinas/genética , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Quinasas Janus/genética , Quinasas Janus/metabolismo , Factor de Transcripción STAT1 , Factor de Transcripción STAT3 , Linfocitos T/metabolismo
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...