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Disbiosis/patología , Microbioma Gastrointestinal/fisiología , Mucosa Intestinal/patología , Quinasa de Cadena Ligera de Miosina/metabolismo , Uniones Estrechas/patología , Animales , Ansiedad/etiología , Ansiedad/psicología , Conducta Animal/fisiología , Modelos Animales de Enfermedad , Disbiosis/complicaciones , Disbiosis/microbiología , Disbiosis/psicología , Femenino , Humanos , Mucosa Intestinal/citología , Mucosa Intestinal/inervación , Mucosa Intestinal/microbiología , Masculino , Ratones Transgénicos , Quinasa de Cadena Ligera de Miosina/genética , Nocicepción/fisiología , PermeabilidadRESUMEN
The regeneration of intestinal epithelial are maintained by continuous differentiation and proliferation of intestinal stem cells (ISCs) under physiological and pathological conditions. However, little is known about the regulatory effect of intestinal microbiota on its recovery ability to repair damaged mucosal barrier. In this study, we established intestinal organoids and lamina propria lymphocytes (LPLs) co-cultured system, plus mice experiments, to explore the protective effect of Lactobacillus reuteri D8 on integrity of intestinal mucosa. We found that only live L. reuteri D8 was effective in protecting the morphology of intestinal organoids and normal proliferation of epithelial stained with EdU under TNF-α treatment, which was also further verified in mice experiments. L. reuteri D8 colonized in the intestinal mucosa and ameliorated intestinal mucosa damage caused by DSS treatment, including improvement of body weight, colon length, pathological change, and proliferation level. The repair process stimulated by L. reuteri D8 was also accompanied with increased numbers of Lgr5+ and lysozyme+ cells both in intestinal organoids and mice intestine. Furthermore, we demonstrated that D8 metabolite indole-3-aldehyde stimulated LPLs to secret IL-22 through aryl hydrocarbon receptor (AhR) and then induced phosphorylation of STAT3 to accelerate proliferation of intestinal epithelial, thus recovering damaged intestinal mucosa. Our findings indicate L. reuteri protects intestinal barrier and activates intestinal epithelial proliferation, which sheds light on treatment approaches for intestinal inflammation based on ISCs with probiotics Lactobacillus and daily probiotic consumption in heath foods.
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Interleucinas/metabolismo , Mucosa Intestinal/metabolismo , Lactobacillus/fisiología , Linfocitos/metabolismo , Factor de Transcripción STAT3/metabolismo , Animales , Técnicas de Cocultivo , Colitis/microbiología , Colitis/patología , Sulfato de Dextran/toxicidad , Indoles/farmacología , Mucosa Intestinal/citología , Mucosa Intestinal/efectos de los fármacos , Linfocitos/citología , Linfocitos/efectos de los fármacos , Ratones , Ratones Endogámicos C57BL , Membrana Mucosa/citología , Muramidasa/metabolismo , Fosforilación , Receptores de Hidrocarburo de Aril/metabolismo , Receptores Acoplados a Proteínas G/metabolismo , Transducción de Señal , Células Madre/citología , Células Madre/metabolismo , Factor de Necrosis Tumoral alfa/farmacología , Interleucina-22RESUMEN
Light-absorbing, atmospheric particles have gained greater attention in recent years because of their direct and indirect impacts on regional and global climate. Atmospheric black carbon (BC) aerosol is a leading climate warming agent, yet uncertainties in the global direct aerosol radiative forcing remain large. Based on a year of aerosol absorption measurements at seven wavelengths, BC concentrations were investigated in Dhanbad, the coal capital of India. Coal is routinely burned for cooking and residential heat as well as in small industries. The mean daily concentrations of ultraviolet-absorbing black carbon measured at 370nm (UVBC) and black carbon measured at 880nm (BC) were 9.8±5.7 and 6.5±3.8µgm-3, respectively. The difference between UVBC and BC, Delta-C, is an indicator of biomass or residential coal burning and averaged 3.29±4.61µgm-3. An alternative approach uses the Ǻngstrom Exponent (AE) to estimate the biomass/coal and traffic BC concentrations. Biomass/coal burning contributed ~87% and high temperature, fossil-fuel combustion contributed ~13% to the annual average BC concentration. The post-monsoon seasonal mean UVBC values were 10.9µgm-3 and BC of 7.2µgm-3. Potential source contribution function analysis showed that in the post-monsoon season, air masses came from the central and northwestern Indo-Gangetic Plains where there is extensive agricultural burning. The mean winter UVBC and BC concentrations were 15.0 and 10.1µgm-3, respectively. These higher values were largely produced by local sources under poor dispersion conditions. The direct radiative forcing (DRF) due to UVBC and BC at the surface (SUR) and the top of the atmosphere (TOA) were calculated. The mean atmospheric heating rates due to UVBC and BC were estimated to be 1.40°Kday-1 and 1.18°Kday-1, respectively. This high heating rate may affect the monsoon circulation in this region.
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In the two inflammatory bowel diseases, ulcerative colitis (UC) and Crohn's disease (CD), altered expression of tight junction (TJ) proteins leads to an impaired epithelial barrier including increased uptake of luminal antigens supporting the inflammation. Here, we focused on regulation of tricellulin (Tric), a protein of the tricellular TJ essential for the barrier against macromolecules, and hypothesized a role in paracellular antigen uptake. We report that Tric is downregulated in UC, but not in CD, and that its reduction increases the passage of macromolecules. Using a novel visualization method, passage sites were identified at TJ regions usually sealed by Tric. We show that interleukin-13 (IL-13), beyond its known effect on claudin-2, downregulates Tric expression. These two effects of IL-13 are regulated by different signaling pathways: The IL-13 receptor α1 upregulates claudin-2, whereas IL-13 receptor α2 downregulates Tric. We suggest to target the α2 receptor in future developments of therapeutical IL-13-based biologicals.
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Colitis Ulcerosa/inmunología , Inflamación/inmunología , Subunidad alfa2 del Receptor de Interleucina-13/metabolismo , Mucosa Intestinal/fisiología , Proteína 2 con Dominio MARVEL/metabolismo , Uniones Estrechas/metabolismo , Adulto , Anciano , Antígenos/inmunología , Antígenos/metabolismo , Claudina-2/metabolismo , Enfermedad de Crohn/inmunología , Regulación hacia Abajo , Femenino , Células HT29 , Humanos , Interleucina-13/metabolismo , Subunidad alfa1 del Receptor de Interleucina-13/metabolismo , Sustancias Macromoleculares/inmunología , Sustancias Macromoleculares/metabolismo , Masculino , Persona de Mediana Edad , Terapia Molecular Dirigida , Transducción de Señal , Adulto JovenRESUMEN
Compromised intestinal barrier function is a prominent feature of inflammatory bowel disease (IBD). However, links between intestinal barrier loss and disease extend much further, including documented associations with celiac disease, type I diabetes, rheumatoid arthritis, and multiple sclerosis. Intestinal barrier loss has also been proposed to have a critical role in the pathogenesis of graft-versus-host disease (GVHD), a serious, potentially fatal consequence of hematopoietic stem cell transplantation. Experimental evidence has begun to support this view, as barrier loss and its role in initiating and establishing a pathogenic inflammatory cycle in GVHD is emerging. Here we discuss similarities between IBD and GVHD, mechanisms of intestinal barrier loss in these diseases, and the crosstalk between barrier loss and the immune system, with a special focus on natural killer (NK) cells. Unanswered questions and future research directions on the topic are discussed along with implications for treatment.
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Enfermedad Injerto contra Huésped/etiología , Enfermedades Inflamatorias del Intestino/etiología , Mucosa Intestinal/patología , Animales , Enfermedad Injerto contra Huésped/metabolismo , Enfermedad Injerto contra Huésped/patología , Enfermedad Injerto contra Huésped/terapia , Humanos , Enfermedades Inflamatorias del Intestino/metabolismo , Enfermedades Inflamatorias del Intestino/patología , Enfermedades Inflamatorias del Intestino/terapia , Mucosa Intestinal/inmunología , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologíaRESUMEN
Addiction to nicotine and the ability to quit smoking are influenced by genetic factors. We used functional genomic approaches (chromatin immunoprecipitation (ChIP) and whole-genome sequencing) to identify cAMP response element-binding protein (CREB) targets following chronic nicotine administration and withdrawal (WD) in rodents. We found that chronic nicotine and WD differentially modulate CREB binding to the gene for neuregulin 3 (NRG3). Quantitative analysis of saline, nicotine and nicotine WD in two biological replicates corroborate this finding, with NRG3 increases in both mRNA and protein following WD from chronic nicotine treatment. To translate these data for human relevance, single-nucleotide polymorphisms (SNPs) across NRG3 were examined for association with prospective smoking cessation among smokers of European ancestry treated with transdermal nicotine in two independent cohorts. Individual SNP and haplotype analysis support the association of NRG3 SNPs and smoking cessation success. NRG3 is a neural-enriched member of the epidermal growth factor family, and a specific ligand for the receptor tyrosine kinase ErbB4, which is also upregulated following nicotine treatment and WD. Mice with significantly reduced levels of NRG3 or pharmacological inhibition of ErbB4 show similar reductions in anxiety following nicotine WD compared with control animals, suggesting a role for NRG3 in nicotine dependence. Although the function of the SNP in NRG3 in humans is not known, these data suggest that Nrg3/ErbB4 signaling may be an important factor in nicotine dependence.
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Predisposición Genética a la Enfermedad/genética , Péptidos y Proteínas de Señalización Intracelular/genética , Neurregulinas/genética , Tabaquismo/genética , Adolescente , Adulto , Afatinib , Anciano , Animales , Conducta Animal/efectos de los fármacos , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , ADN/metabolismo , Femenino , Estudio de Asociación del Genoma Completo , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Humanos , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Masculino , Ratones , Persona de Mediana Edad , Mutación , Neurregulinas/metabolismo , Nicotina/farmacología , Polimorfismo de Nucleótido Simple/genética , Quinazolinas/farmacología , Ratas , Receptor ErbB-4/antagonistas & inhibidores , Fumar/tratamiento farmacológico , Fumar/genética , Síndrome de Abstinencia a Sustancias/genética , Dispositivos para Dejar de Fumar Tabaco , Población Blanca/genética , Adulto JovenRESUMEN
The Chagos Archipelago was designated a no-take marine protected area (MPA) in 2010; it covers 550 000 km2, with more than 60 000 km2 shallow limestone platform and reefs. This has doubled the global cover of such MPAs.It contains 25-50% of the Indian Ocean reef area remaining in excellent condition, as well as the world's largest contiguous undamaged reef area. It has suffered from warming episodes, but after the most severe mortality event of 1998, coral cover was restored after 10 years.Coral reef fishes are orders of magnitude more abundant than in other Indian Ocean locations, regardless of whether the latter are fished or protected.Coral diseases are extremely low, and no invasive marine species are known.Genetically, Chagos marine species are part of the Western Indian Ocean, and Chagos serves as a 'stepping-stone' in the ocean.The no-take MPA extends to the 200 nm boundary, and. includes 86 unfished seamounts and 243 deep knolls as well as encompassing important pelagic species.On the larger islands, native plants, coconut crabs, bird and turtle colonies were largely destroyed in plantation times, but several smaller islands are in relatively undamaged state.There are now 10 'important bird areas', coconut crab density is high and numbers of green and hawksbill turtles are recovering.Diego Garcia atoll contains a military facility; this atoll contains one Ramsar site and several 'strict nature reserves'. Pollutant monitoring shows it to be the least polluted inhabited atoll in the world. Today, strict environmental regulations are enforced.Shoreline erosion is significant in many places. Its economic cost in the inhabited part of Diego Garcia is very high, but all islands are vulnerable.Chagos is ideally situated for several monitoring programmes, and use is increasingly being made of the archipelago for this purpose.
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The prevalence of obesity, a major risk factor for many chronic diseases, has risen in most developed countries over the past several decades. The economic burden for both public and private health care systems is substantial. Although certain non-pharmaceutical interventions have been proven efficacious in specific populations, the lack of scalability has caused many of these programmes to fail in sustainably decreasing the percent of patients who are overweight or obese. The benefits of other interventions, such as pharmaceutical agents, medical devices and surgery, should therefore be carefully considered: this article focuses on the first of these strategies. Various pharmaceutical products have been plagued with safety concerns or patient non-adherence because of unpleasant side effects. Therefore, the need for additional antiobesity drugs that are both safe and effective is considerable. This article discusses the regulatory landscape for the development of new antiobesity compounds in the United States and Europe and considers the ramifications of greater or lesser regulatory burdens.
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Fármacos Antiobesidad/uso terapéutico , Depresores del Apetito/uso terapéutico , Obesidad/tratamiento farmacológico , Fármacos Antiobesidad/economía , Depresores del Apetito/economía , Análisis Costo-Beneficio , Femenino , Humanos , Masculino , Obesidad/complicaciones , Obesidad/economía , Salud Pública , Factores de Riesgo , Conducta de Reducción del RiesgoRESUMEN
The Cl/HCO(3) exchanger downregulated in adenoma (DRA) mediates electroneutral NaCl absorption in the intestine together with the apical Na/H exchanger NHE3. Lipid rafts (LR) modulate transport activity and are involved in phosphatidylinositol 3-kinase (PI3-kinase)-dependent trafficking of NHE3. Although DRA and NHE3 interact via PDZ adaptor proteins of the NHERF family, the role of LR and PDZ proteins in the regulation of DRA is unknown. We examined the association of DRA with LR using the nonionic detergent Triton X-100. DRA cofractionated with LR independently of its PDZ binding motif. Furthermore, DRA interacts with PDZK1, E3KARP, and IKEPP in LR, although their localization within lipid rafts is independent of DRA. Disruption of LR integrity resulted in the disappearance of DRA from LR, in a decrease of its surface expression and in a reduction of its activity. In HEK cells the inhibition of DRA by LR disruption was entirely dependent on the presence of the PDZ interaction motif. In addition, in Caco-2/BBE cells the inhibition by LR disruption was more pronounced in wild-type DRA than in mutated DRA (DRA-ETKFminus; lacking the PDZ binding motif)-expressing cells. Inhibition of PI3-kinase decreased the activity and the cell surface expression of wild-type DRA but not of DRA-ETKFminus; the partitioning into LR was unaffected. Furthermore, simultaneous inhibition of PI3-kinase and disruption of LR did not further decrease DRA activity and cell surface expression compared with LR disruption only. These results suggest that the activity of DRA depends on its LR association, on its PDZ interaction, and on PI3-kinase activity.
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Antiportadores/metabolismo , Microdominios de Membrana/fisiología , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfoproteínas/metabolismo , Transporte de Proteínas/fisiología , Intercambiadores de Sodio-Hidrógeno/metabolismo , Animales , Antiportadores/genética , Línea Celular , Antiportadores de Cloruro-Bicarbonato , Colesterol/metabolismo , Regulación Neoplásica de la Expresión Génica/fisiología , Humanos , Familia de Multigenes , Fosfoproteínas/genética , Intercambiadores de Sodio-Hidrógeno/genética , Transportadores de SulfatoRESUMEN
We investigate neutral evolution during range shifts in a strategic model of a metapopulation occupying a climate gradient. Using heritable, neutral markers, we track the spatio-temporal fate of lineages. Owing to iterated founder effects ('mutation surfing'), survival of lineages derived from the leading range limit is enhanced. At trailing limits, where habitat suitability decreases, survival is reduced (mutations 'wipe out'). These processes alter (i) the spatial spread of mutations, (ii) origins of persisting mutations and (iii) the generation of diversity. We show that large changes in neutral evolution can be a direct consequence of range shifting.
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Evolución Biológica , Demografía , Efecto Invernadero , Modelos Biológicos , Extinción Biológica , Variación Genética , MutaciónRESUMEN
The decline in industrial melanism over the last quarter century constitutes an exceptional case of an evolutionary change, varying in both time and space, and between species. In Biston betularia and Odontoptera bidentata, the change in melanic frequency is closely replicated at two sites 0.5 km apart. Between seven sites 50-100 km apart, there is heterogeneity in both the speed and timing of change. At sites that were heavily industrialized, the change is faster, from an initially higher frequency, and starts later than at sites which are more rural.We propose a method for estimating systematic change during sigmoid declines in melanic frequencies. This fails to show any significant change over time in selective coefficients. It is concluded that the overall pattern of change has been driven largely by events in the most polluted and industrialized parts of the country. Although migration may contribute to the estimated selective values, natural selection is the only credible explanation for the overall decline.
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Melaninas/genética , Mariposas Nocturnas/genética , Animales , Genética de Población , Industrias , Pigmentación , Selección GenéticaAsunto(s)
Enfermedades Inflamatorias del Intestino/etiología , Mucosa Intestinal/fisiopatología , Permeabilidad de la Membrana Celular/fisiología , Colitis Ulcerosa/etiología , Colitis Ulcerosa/fisiopatología , Enfermedad de Crohn/etiología , Enfermedad de Crohn/fisiopatología , Epitelio/fisiopatología , Humanos , Enfermedades Inflamatorias del Intestino/fisiopatología , Interferón gamma/metabolismo , Proteínas de la Membrana/metabolismo , Modelos Biológicos , Quinasa de Cadena Ligera de Miosina/metabolismo , Uniones Estrechas/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
BACKGROUND: It has previously been reported that the risk of ventricular arrhythmias is positively associated with ambient air pollution among patients with implantable cardioverter defibrillators (ICD) in Boston. AIMS: To assess the association of community exposures to air pollution with ventricular arrhythmias in a cohort of ICD patients in metropolitan St Louis, Missouri. METHODS: ICD detected episodes reported during clinical follow up were abstracted and reviewed by an electrophysiologist to identify ventricular arrhythmias. A total of 139 ventricular arrhythmias were identified among 56 patients. A case-crossover design was used with control periods matched on weekday and hour of the day within the same calendar month. Conditional logistic regression models were adjusted for temperature, barometric pressure, and relative humidity in the 24 hours preceding the event. RESULTS: There was a significant (24%, 95% CI 7% to 44%) increase in risk of ventricular arrhythmias associated with each 5 ppb increase in mean sulphur dioxide and non-significantly increased risk (22%, 95% CI -6% to 60%; and 18%, 95% CI -7% to 50%) associated with increases in nitrogen dioxide (6 ppb) and elemental carbon (0.5 microg/m3), respectively in the 24 hours before the arrhythmia. CONCLUSIONS: These results provide evidence of an association between ventricular arrhythmias and ambient air pollutants in St Louis. This is consistent with previous results from Boston, although the pollutants responsible for the increased risk are different.
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Contaminantes Atmosféricos/toxicidad , Contaminación del Aire/efectos adversos , Arritmias Cardíacas/epidemiología , Desfibriladores Implantables , Adulto , Anciano , Anciano de 80 o más Años , Contaminantes Atmosféricos/análisis , Estudios de Cohortes , Estudios Cruzados , Femenino , Humanos , Masculino , Persona de Mediana Edad , Missouri/epidemiología , Dióxido de Nitrógeno/efectos adversos , Dióxido de Nitrógeno/análisis , Tamaño de la Partícula , Dióxido de Azufre/análisis , Dióxido de Azufre/toxicidadRESUMEN
We previously developed a swine animal model in which natural host resistance to Campylobacter jejuni is altered by experimental infection with low numbers of the nematode Trichuris suis. Pigs naturally colonized with C. jejuni experience colitis because of the invasion of the bacterium approximately 21 days after exposure to T. suis. To better understand the mechanism of T. suis-dependent C. jejuni colitis, we evaluated the effects of T. suis excretory-secretory products (ESPs) on intestinal epithelial cells (IECs) and the influence of ESP on C. jejuni invasion in IECs under in vitro conditions. Viability assays revealed a dose-dependent cytotoxic response in ESP-treated IECs, particularly IPEC-1 and INT407 cells. Transepithelial electrical resistance dropped significantly in IPEC-1 cells treated on apical and basolateral surfaces, but not in those treated only on apical surfaces. Using the gentamicin-killing assay, reduced numbers of intracellular C. jejuni were recovered from IECs treated with ESP at 1 mg protein/ml concentration. This observation can be at least partially explained by a novel antibacterial activity in ESP. Contrary to our hypothesis, ESP at subtoxic concentrations did not enhance invasion. In addition to mechanical damage from worms, these results suggest that soluble products released by T. suis contribute to IEC damage at the site of worm attachment.
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Campylobacter jejuni/patogenicidad , Mucosa Intestinal/microbiología , Mucosa Intestinal/parasitología , Trichuris/fisiología , Animales , Campylobacter jejuni/efectos de los fármacos , Campylobacter jejuni/crecimiento & desarrollo , Línea Celular , Supervivencia Celular/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Células Epiteliales/microbiología , Células Epiteliales/parasitología , Humanos , Mucosa Intestinal/efectos de los fármacos , Modelos Animales , Porcinos , Extractos de Tejidos/farmacología , Trichuris/metabolismoRESUMEN
BACKGROUND: Early enteral nutrition in patients following traumatic injury is an important intervention. However, after shock-resuscitation, intestinal hypoperfusion persists despite adequate systemic resuscitation. Our previous in vivo rat studies indicate that hypoperfusion impairs mucosal function in the small intestine. Therefore, the current study sought to improve previous in vitro models by the following means: (1) We used Caco-2 monolayers stably transfected with the brush-border sodium-glucose co-transporter (SGLT-1); and (2) we created an environment that mimicked the physiologic enterocyte environment. We hypothesized that hypoxic alterations of epithelial function in an in vitro model are comparable to those of an in vivo rat model. METHODS: After 21 days, monolayers were randomized to receive 24 hours of incubation in a normoxic or hypoxic environment. Cells were further randomized to receive 1 of 4 nutrient treatments: mannitol (an osmotic control), glucose (uses SGLT-1 and is metabolized), 3-O-methylglucose (3-O-mg; uses SGLT-1 and is not metabolized), or fructose (does not use SGLT-1 but can be metabolized). RESULTS: Transepithelial resistance (p = .007) and short-circuit current (p = .05) were lower in hypoxic groups. When compared with normoxic groups, hypoxic groups had significantly impaired glucose (p < .001) but not glutamine transport, irrespective of nutrient treatment. Additionally, adenosine triphosphate/adenosine diphosphate ratio was reduced (p = .01) and lactate concentration was increased (p < .001) during hypoxia. CONCLUSIONS: In summary, results from this in vitro study using Caco-2BBe cells stably transfected with SGLT-1 correspond to results obtained in the in vivo rat model. Therefore, this is an appropriate in vitro model in which to study cellular alterations caused by the hypoxic small intestine, with the goal of ensuring safe early enteral nutrition following traumatic injury.
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Hipoxia de la Célula , Nutrición Enteral , Enterocitos/fisiología , Modelos Biológicos , 3-O-Metilglucosa/administración & dosificación , Adenosina Difosfato/análisis , Adenosina Trifosfato/análisis , Animales , Transporte Biológico/efectos de los fármacos , Células CACO-2 , Conductividad Eléctrica , Impedancia Eléctrica , Enterocitos/química , Epitelio/fisiología , Fructosa/administración & dosificación , Glucosa/administración & dosificación , Glucosa/metabolismo , Glutamina/metabolismo , Humanos , Ácido Láctico/análisis , Manitol/administración & dosificación , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/fisiología , Proteínas de Transporte de Monosacáridos/genética , Proteínas de Transporte de Monosacáridos/fisiología , Ratas , Sodio/farmacología , Transportador 1 de Sodio-Glucosa , TransfecciónRESUMEN
Na(+)- nutrient cotransport-dependent regulation of paracellular permeability has been demonstrated in rodent intestine and human intestinal epithelial cell lines. In cell lines this regulation is associated with phosphorylation of myosin II regulatory light chain (MLC). However, the subcellular localization of phosphorylated MLC during this regulation has not been studied and regulation of paracellular permeability and MLC phosphorylation has not been studied in isolated human intestine. To evaluate these events in human jejunum, isolated mucosa was mounted in Ussing chambers, characterized electrophysiologically, and then immunostained using anti-phosphorylated MLC and anti-total MLC antisera. MLC phosphorylation was assessed by calculating the ratio of anti-phosphorylated MLC signal to anti-total MLC signal within defined regions. Transmucosal resistance of mucosae without active Na(+)-glucose cotransport was 37 +/- 3% greater than that of mucosae with active Na(+)-glucose cotransport within 15 min. Quantitative double-label immunofluorescence showed that the phosphorylated MLC-to-total MLC ratio increased by 45 +/- 4% within the perijunctional actomyosin ring when Na(+)-glucose cotransport was active. Thus regulation of transmucosal resistance by Na(+)-glucose cotransport is accompanied by increased MLC phosphorylation within the perijunctional actomyosin ring. These data support the proposed critical role of the perijunctional cytoskeleton in physiological regulation of human small intestinal paracellular permeability.
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Glucosa/metabolismo , Mucosa Intestinal/fisiología , Glicoproteínas de Membrana/fisiología , Proteínas de Transporte de Monosacáridos/fisiología , Cadenas Ligeras de Miosina/metabolismo , Sodio/metabolismo , Actomiosina , Transporte Biológico , Permeabilidad de la Membrana Celular , Impedancia Eléctrica , Técnica del Anticuerpo Fluorescente , Humanos , Mucosa Intestinal/ultraestructura , Yeyuno/fisiología , Microscopía Fluorescente , Fosforilación , Transportador 1 de Sodio-Glucosa , Uniones EstrechasRESUMEN
Cytoplasmic pH (pH(i)) was evaluated during Na(+)-glucose cotransport in Caco-2 intestinal epithelial cell monolayers. The pH(i) increased by 0.069 +/- 0.002 within 150 s after initiation of Na(+)-glucose cotransport. This increase occurred in parallel with glucose uptake and required expression of the intestinal Na(+)-glucose cotransporter SGLT1. S-3226, a preferential inhibitor of Na(+)/H(+) exchanger (NHE) isoform 3 (NHE3), prevented cytoplasmic alkalinization after initiation of Na(+)-glucose cotransport with an ED(50) of 0.35 microM, consistent with inhibition of NHE3, but not NHE1 or NHE2. In contrast, HOE-694, a poor NHE3 inhibitor, failed to significantly inhibit pH(i) increases at <500 microM. Na(+)-glucose cotransport was also associated with activation of p38 mitogen-activated protein (MAP) kinase, and the p38 MAP kinase inhibitors PD-169316 and SB-202190 prevented pH(i) increases by 100 +/- 0.1 and 86 +/- 0.1%, respectively. Conversely, activation of p38 MAP kinase with anisomycin induced NHE3-dependent cytoplasmic alkalinization in the absence of Na(+)-glucose cotransport. These data show that NHE3-dependent cytoplasmic alkalinization occurs after initiation of SGLT1-mediated Na(+)-glucose cotransport and that the mechanism of this NHE3 activation requires p38 MAP kinase activity. This coordinated regulation of glucose (SGLT1) and Na(+) (NHE3) absorptive processes may represent a functional activation of absorptive enterocytes by luminal nutrients.
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Citoplasma/enzimología , Mucosa Intestinal/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Transporte de Monosacáridos/metabolismo , Intercambiadores de Sodio-Hidrógeno/metabolismo , Actomiosina/metabolismo , Western Blotting , Células CACO-2 , Citoplasma/efectos de los fármacos , Inhibidores Enzimáticos/farmacología , Células Epiteliales/efectos de los fármacos , Células Epiteliales/metabolismo , Epitelio/efectos de los fármacos , Epitelio/metabolismo , Humanos , Concentración de Iones de Hidrógeno , Mucosa Intestinal/citología , Mucosa Intestinal/efectos de los fármacos , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Transportador 1 de Sodio-Glucosa , Intercambiador 3 de Sodio-Hidrógeno , Proteínas Quinasas p38 Activadas por MitógenosRESUMEN
African Americans as a group have a higher incidence of chronic hepatitis C (CHC) than Caucasians but are often under-represented in clinical trials used to define response rates to interferon therapy. The aim of this study was to compare African Americans with Caucasians with respect to end-of-treatment response to interferon. This retrospective study had 61 African Americans and 49 Caucasians with CHC. All patients were treated for at least 12 weeks with interferon-alpha2b (Intron A) thrice weekly. End-of-treatment response was defined as three consecutive nondetectable HCV RNA measurements at least 1 month apart. Sustained response was defined as a negative serum HCV RNA 6 months after end of treatment. Of the 110 patients, 19 achieved an end-of-treatment response (17%) but only four achieved a sustained response (4/110=4%). Of the patients achieving a sustained response, one was genotype 1 (male Caucasian), three were genotype 2/3 with four patients having no follow-up information. The end-of-treatment response was 7% for patients with genotype 1 and 71% for genotype non-1 (P < 0.005 for genotype non-1). The end-of-treatment response was significantly higher in Caucasians (14/49=31%) compared with African Americans (5/61=8%; P < 0.05). A lower response rate in African Americans with genotype 1 in contrast to Caucasians was the primary reason for the difference in end-of-treatment response (1/45=2% vs. 5/33=15%, P < 0.05). Hence, interferon treatment resulted in a poor sustained response rate in the group of patients representative of the urban populations with the highest prevalence of hepatitis C. A genotype other than type 1 was the strongest predictor of end-of-treatment response in patients treated but over 86% of patients in this urban clinic were genotype 1. Caucasians were more likely to respond than African Americans, especially in patients with genotype 1.