RESUMEN
We investigated the potential of mouse embryonic stem (ES) cells to differentiate into hepatocytes in vitro. Differentiating ES cells expressed endodermal-specific genes, such as alpha-fetoprotein, transthyretin, alpha 1-anti-trypsin and albumin, when cultured without additional growth factors and late differential markers of hepatic development, such as tyrosine aminotransferase (TAT) and glucose-6-phosphatase (G6P), when cultured in the presence of growth factors critical for late embryonic liver development. Further, induction of TAT and G6P expression was induced regardless of expression of the functional SEK1 gene, which is thought to provide a survival signal for hepatocytes during an early stage of liver morphogenesis. The data indicate that the in vitro ES differentiation system has a potential to generate mature hepatocytes. The system has also been found useful in analyzing the role of growth factors and intracellular signaling molecules in hepatic development.
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Hepatocitos/citología , Hígado/citología , Hígado/embriología , MAP Quinasa Quinasa 4 , Células Madre/citología , Células Madre/metabolismo , Animales , Antígenos de Diferenciación/biosíntesis , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/fisiología , Línea Celular , Linaje de la Célula , Colágeno/metabolismo , Matriz Extracelular/metabolismo , Sustancias de Crecimiento/farmacología , Hepatocitos/efectos de los fármacos , Ratones , Quinasas de Proteína Quinasa Activadas por Mitógenos/deficiencia , Quinasas de Proteína Quinasa Activadas por Mitógenos/genética , Quinasas de Proteína Quinasa Activadas por Mitógenos/metabolismo , ARN Mensajero/metabolismo , Células Madre/efectos de los fármacosRESUMEN
BACKGROUND/PURPOSE: There are few long-term chronological reviews examining the incidence of total parenteral nutrition (TPN)-associated intrahepatic cholestasis (TPNAC) in infants. The authors therefore reviewed TPNAC in their 25-year series, and also looked at the current problems associated with TPN in infants. METHODS: Two hundred seventy-three surgical neonates who received TPN for more than 2 weeks were divided into 3 groups chronologically: group A (1971 through 1982, n = 77), group B (1983 through 1987, n = 72), and group C (1992 through 1996, n = 124). TPNAC was defined as serum direct bilirubin (DB) level greater than 2.0 mg/dL during the neonatal period. RESULTS: The incidence of TPNAC in groups A, B and C was 57%, 31%, and 25% (P< .01), respectively, and the mortality rate from TPN-associated complications was 13%, 3%, and 3% (P< .05), respectively. Over the last 5 years, severe TPNAC developed in 20 patients (16%). Four of 20 died of TPN-associated sepsis with hepatic failure; 2 had hypoganglionosis with intractable stagnant enteritis and subsequent sepsis, and 2 had fatal respiratory or cardiac disease. CONCLUSIONS: The incidence of TPNAC in surgical neonates and TPN-associated mortality rates have decreased significantly. The mortality rate, however, still remains at 3%. Two of 4 fatal cases had hypoganglionosis, which were totally dependent on TPN. In patients who require long-term TPN, TPN still has unsolved problems, and small bowel transplantation may be indicated.
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Colestasis Intrahepática/epidemiología , Nutrición Parenteral Total/efectos adversos , Colestasis Intrahepática/etiología , Humanos , Incidencia , Recién Nacido , Factores de TiempoRESUMEN
L-Asparaginase is widely used in the treatment of acute lymphoblastic leukemia. L-Asparaginase preparation derived from E. coli converts asparagine (Asn) and glutamine (Gln) to aspartate (Asp) and glutamate (Glu), respectively, and causes rapid depletion of Asn and Gln. It thus suppresses growth of malignant cells that are more dependent on an exogenous source of Asn and Gln than are normal cells. It remains unclear, however, which signaling events in leukemic cells are affected by L-asparaginase. Recently, amino acid sufficiency has been demonstrated to selectively regulate p70 S6 kinase (p70(s6k)) and eukaryotic initiation factor 4E-binding protein 1 (4E-BP1), both of which are targeted by the anti-proliferative drug rapamycin. Here we demonstrate that addition of L-asparaginase to human leukemic cells inhibits activity of p70(s6k) and phosphorylation of 4E-BP1, but not activities of other cell growth-related serine/threonine kinases. The rate and kinetics of p70(s6k) inhibition by L-asparaginase were comparable to those seen by deprivation of Asn and/or Gln from cell culture media, suggesting that the effect of L-asparaginase on p70(s6k) is explained by depletion of Asn and/or Gln. Moreover, L-Asparaginase as well as rapamycin selectively suppressed synthesis of ribosomal proteins at the level of mRNA translation. These data indicate that L-asparaginase and rapamycin target a common signaling pathway in leukemic cells.
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Asparaginasa/farmacología , Proteínas Portadoras , Transducción de Señal/efectos de los fármacos , Sirolimus/antagonistas & inhibidores , Proteínas Adaptadoras Transductoras de Señales , Proteínas de Ciclo Celular , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Humanos , Fosfoproteínas/antagonistas & inhibidores , Fosforilación , Biosíntesis de Proteínas/efectos de los fármacos , Proteínas Serina-Treonina Quinasas/antagonistas & inhibidores , ARN Mensajero/genética , Proteínas Quinasas S6 Ribosómicas/antagonistas & inhibidores , Proteínas Quinasas S6 Ribosómicas/metabolismo , Proteínas Ribosómicas/genética , Sirolimus/farmacología , Células Tumorales CultivadasRESUMEN
BACKGROUND: The effects of N-acetylcysteine (NAC) as a mucolytic agent on the uptake of fluorescent polystyrene microparticles by Peyer's patches, on intestinal permeability, and on subsequent transport to mesenteric lymph nodes (MLNs) were investigated to establish the role of mucus gel layer in this process. METHODS: Twenty rats were divided into two groups: control (n = 10) and NAC (n = 10). Fluorescent polystyrene latex beads of 3.2+/-0.2 microm in diameter were used as a probe for measuring the previously mentioned parameters. The solution of latex beads (0.1 mL) was injected into a 2-cm length of ileal loop containing Peyer's patches, with 0.1 mL of saline (control group) or with 0.1 mL of NAC solution (NAC group) within 10 cm proximal from the ileocaecal valve. Intestinal loops, portal blood, and neighboring MLNs were taken within 1 hour of injection. Intestinal sections were stained by periodic acid-Schiff reagent. Peyer's patches and MLNs were analyzed for the count of particles by image analysis using a confocal laser scanning microscope. RESULTS: Morphologically, periodic acid-Schiff positive uniform mucus gel was present in front of Peyer's patches of the control group, and mucus gel layer was disrupted and noncontinuous in the NAC group. The number of particles within Peyer's patches and MLNs in the NAC group was significantly higher than that in the control group (p<.001). Intestinal permeability of latex beads in the NAC group was significantly higher than that in the control group (p<.001). CONCLUSIONS: These data suggest that the mucus gel layer located in front of Peyer's patches is one of the important factors for the uptake of noxious macromolecules, and this in turn plays a major role on small intestinal permeability and subsequent translocation to MLNs.
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Acetilcisteína/farmacología , Mucosa Intestinal/efectos de los fármacos , Ganglios Linfáticos Agregados/efectos de los fármacos , Animales , Permeabilidad de la Membrana Celular/efectos de los fármacos , Depuradores de Radicales Libres , Íleon/efectos de los fármacos , Íleon/metabolismo , Mucosa Intestinal/metabolismo , Ganglios Linfáticos/metabolismo , Masculino , Microesferas , Permeabilidad/efectos de los fármacos , Ganglios Linfáticos Agregados/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Effect of supplemental alanyl-glutamine in standard TPN (S-TPN) on luminal mucus gel and small intestinal permeability was investigated. METHODS: Thirty Sprague-Dawley rats were divided into group I (n = 10), receiving standard rat diet; group II (n = 10), receiving S-TPN; and group III (n = 10), receiving alanyl-glutamine-supplemented TPN for 1 week. After 1 week, fluorescein isothiocyanate (FITC)-dextran was injected into the small intestine of the rats, and they were killed. A small intestinal sample and portal blood were obtained for morphologic and functional analysis of mucus gel and intestinal permeability. RESULTS: In group II, thickness and optical density of mucus gel per millimeter serosal length of intestine were significantly lower than group I (p<.001) and were significantly higher in group III than in group II (p<.001). The number of goblet cells in the villi and in the crypt of the small intestine was significantly lower in group II than in group I (p<.001) and was significantly higher in group III than in group II (p<.001), with the exception of the villi of jejunum. Villous and crypt surface area per millimeter serosal length of intestine was significantly lower in group II than in group I (p<.001) and was significantly higher in group III than in group II (p<.001). Small intestinal permeability to FITC-dextran was significantly higher in group II than in group I (p<.001) and was significantly lower in group III than in group II (p<.001). Glucosamine synthetase level was significantly higher in group III than in group I and ileum of group II (p<.001). CONCLUSIONS: Alanyl-glutamine-supplemented TPN prevents a decrease in mucus gel and an increase in small intestinal permeability associated with S-TPN.
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Dipéptidos/farmacología , Mucosa Intestinal/efectos de los fármacos , Intestino Delgado/efectos de los fármacos , Nutrición Parenteral Total , Animales , Fluoresceína-5-Isotiocianato/metabolismo , Colorantes Fluorescentes/metabolismo , Intestino Delgado/metabolismo , Masculino , Permeabilidad/efectos de los fármacos , Ratas , Ratas Sprague-DawleyRESUMEN
In human T-lymphoblastoid cells, downstream signaling events of mammalian target of rapamycin (mTOR), including the activity of p70(s6k) and phosphorylation of eukaryotic initiation factor 4E-binding protein 1, were dependent on amino acid concentration in the culture media, whereas other growth-related protein kinases were not. Amino acid-induced p70(s6k) activation was completely inhibited by rapamycin but only partially inhibited by wortmannin. Moreover, amino acid concentration similarly affected the p70(s6k) activity, which was dependent on a rapamycin-resistant mutant (S2035I) of mTOR. These data indicate that mTOR is required for amino acid-dependent activation of p70(s6k). The mechanism by which amino acids regulate p70(s6k) activity was further explored: 1) amino acid alcohols, which inhibit aminoacylation of tRNA by their competitive binding to tRNA synthetases, suppressed p70(s6k) activity; 2) suppression of p70(s6k) by amino acid depletion was blocked by cycloheximide or puromycin, which inhibit utilization of aminoacylated tRNA in cells; and 3) in cells having a temperature-sensitive mutant of histidyl tRNA synthetase, p70(s6k) was suppressed by a transition of cells to a nonpermissible temperature, which was partially restored by addition of high concentrations of histidine. These results indicate that suppression of tRNA aminoacylation is able to inhibit p70(s6k) activity. Deacylated tRNA may be a factor negatively regulating p70(s6k).
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Aminoácidos/fisiología , Aminoacil-ARN de Transferencia/metabolismo , Proteínas Quinasas S6 Ribosómicas/metabolismo , Androstadienos/farmacología , Animales , Células CHO , Cricetinae , Activación Enzimática , Inhibidores Enzimáticos/farmacología , Humanos , Células Jurkat , Extensión de la Cadena Peptídica de Translación/efectos de los fármacos , Fosforilación , Proteínas Quinasas S6 Ribosómicas/antagonistas & inhibidores , Sirolimus/farmacología , WortmaninaRESUMEN
This study investigated whether interleukin-1alpha-induced metallothionein gene expression is affected by zinc deficiency. Weaning male rats were fed a zinc-deficient (ZD) diet (2 mg zinc/kg) or a zinc-supplemented diet [50.8 mg zinc/kg; controls for the diet included pair-fed (PF) and ad libitum consumption groups (AL)] for 4 wk. All rats except those that served as controls for interleukin-1alpha administration, (injected with vehicle and killed at 0 h) were then injected subcutaneously with interleukin-1alpha (2 x 10(7) units/kg body wt) and killed at 3, 6, 12, 24 and 72 h after the injection. Compared with AL and/or PF rats, zinc depletion significantly reduced zinc concentrations in plasma and liver but not in kidney or intestine, and significantly reduced hepatic, renal, and intestinal metallothionein-1 mRNA levels analyzed by competitive reverse transcription-polymerase chain reaction (RT-PCR). Interleukin-1alpha injection reduced plasma zinc concentration and enhanced liver zinc concentration, but did not affect zinc levels in kidney or intestine. Metallothionein-1 mRNA was significantly elevated by interleukin-1alpha in liver, kidney and intestine of all groups; the levels in liver and kidney of ZD rats 6 h after the injection were significantly higher than those of AL or PF rats. Liver metallothionein protein levels were enhanced after interleukin-1alpha injection in both AL and ZD rats. Semiquantitative RT-PCR revealed significantly higher hepatic levels of interleukin-1 receptor type-I mRNA in ZD rats than in AL and PF rats but no differences in renal or intestinal tissues among groups before interleukin-1alpha challenge. In conclusion, zinc deficiency induces upregulation of metallothionein-1 gene expression in response to interleukin-1alpha challenge in rats.
Asunto(s)
Expresión Génica , Interleucina-1/farmacología , Metalotioneína/genética , Zinc/deficiencia , Animales , Western Blotting , Dieta , Mucosa Intestinal/metabolismo , Riñón/metabolismo , Hígado/metabolismo , Masculino , Metalotioneína/biosíntesis , Metalotioneína/metabolismo , Reacción en Cadena de la Polimerasa , ARN Mensajero/metabolismo , ADN Polimerasa Dirigida por ARN , Ratas , Ratas Sprague-Dawley , Zinc/administración & dosificación , Zinc/metabolismoRESUMEN
Synthesis of inducible nitric oxide synthase (iNOS) in the intestine may result in local tissue damage. We investigated whether a challenge with interleukin-1alpha could give rise to intestinal iNOS expression and diarrhea in rats of differing zinc status. Weaning male rats were fed a zinc-deficient (ZD) diet (2 mg zinc/kg) for 4 wk to induce zinc deficiency or a zinc-supplemented diet [50.8 mg zinc/kg; controls, including pair-fed (PF ) and ad libitum (AL) consumption groups], and then subcutaneously injected with interleukin-1alpha (2 x 10(7) units/kg body wt). Without the interleukin-1alpha challenge, ZD rats had significantly lower plasma zinc concentration than the other groups. Intestinal metallothionein-1 mRNA abundance was lower in ZD rats than in AL rats. iNOS was expressed in the intestine of ZD rats but not in the others. None of the rats experienced diarrhea during the feeding period. Interleukin-1alpha led to a reduction in plasma zinc concentration, enhancement in intestinal metallothionein-1 mRNA levels, and expression of the intestinal iNOS gene in all groups. However, the abundance of iNOS mRNA was significantly higher in ZD rats than in the other groups. The presence of iNOS protein was demonstrated by immunohistochemical staining in the intestine of ZD rats that had been treated with interleukin-1alpha 12 h earlier. In addition, diarrhea occurred in most of the ZD rats and some of the PF rats but not in AL rats after interleukin-1alpha treatment. We conclude that ZD rats respond to interleukin-1alpha challenge more severely than controls, reflected by a more marked and prolonged iNOS expression and a greater incidence of diarrhea.
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Diarrea/etiología , Interleucina-1/farmacología , Mucosa Intestinal/metabolismo , Intestinos/efectos de los fármacos , Óxido Nítrico Sintasa/biosíntesis , Zinc/deficiencia , Animales , Dieta , Regulación Enzimológica de la Expresión Génica , Interleucina-1/efectos adversos , Intestinos/patología , Masculino , Metalotioneína/metabolismo , Óxido Nítrico Sintasa/genética , Ratas , Ratas Sprague-Dawley , Zinc/sangreRESUMEN
Occlusion of central venous access devices (CVADs) is not an uncommon problem duringlong-term parenteral nutrition. A number of techniques have been developed to deal with obstructed CVADs. This study investigated the effectiveness of the sodium hydroxide (NaOH) lock method for gradual CVAD occlusion. When a progressively declining flow was noticed, 0.1 N NaOH solution was injected into the CVAD and locked. Nineteen CVAD occlusions in 11 home parenteral nutrition patients were treated Sixteen of 19 trials cleared the occlusions, whereas 3 of 19 failed. One of the failures was due to a mechanical occlusion and the other two were able to be restored by using ethanol. There were no significant complications. The benefits of this method are: (1) a shorter treatment time and a lower dose than NaOH infusion therapy, (2) it does not require hospital admission and (3) it does not result in bursting of the catheter.
RESUMEN
BACKGROUND: Growth hormone (GH) improves net protein anabolism and stimulates wound healing. Although GH is also known to exert the trophic effect on the intestinal tract, its role in the healing of intestinal ulceration is not known. The aim of this study was to evaluate the effects of exogenous GH coinfused with parenteral nutrition (PN) in an experimental model of inflammatory bowel disease in rats. METHODS: All rats underwent central venous cannulation and were randomized to two groups after induction of small intestinal ulceration with indomethacin. Both groups received the same PN formula. In addition, the GH group (n = 10) received subcutaneous injections of human GH at a dose of 1.0 IU/kg daily for 4 days, whereas the control group (n = 10) received injections of normal saline solution. Nitrogen balance, macroscopic inflammation score, intestinal myeloperoxidase activity, DNA content, and mucosal permeability were determined for each rat. Insulin-like growth factor-I (IGF-I) mRNA was detected by reverse transcription and polymerase chain reaction. RESULTS: Administration of GH significantly improved the cumulative nitrogen balance, ameliorated the gross inflammation score, and decreased intestinal myeloperoxidase activity. Similarly, intestinal permeability was significantly decreased in the GH group as compared with the control group. GH treatment resulted in increased plasma concentration of IGF-I and IGF-I mRNA expressions in both the liver and the small intestine compared with those in the control group. CONCLUSIONS: Exogenous GH plays an important role in accelerating intestinal healing in an experimental model of small bowel ulceration in rats. The mechanisms may include the stimulated IGF-I production, which thereafter augments intestinal epithelial cell growth.
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Hormona de Crecimiento Humana/uso terapéutico , Enfermedades Inflamatorias del Intestino/terapia , Nutrición Parenteral , Animales , Gliceraldehído-3-Fosfato Deshidrogenasas/genética , Humanos , Enfermedades Inflamatorias del Intestino/metabolismo , Factor I del Crecimiento Similar a la Insulina/análisis , Factor I del Crecimiento Similar a la Insulina/genética , Mucosa Intestinal/metabolismo , Masculino , Nitrógeno/metabolismo , ARN Mensajero/análisis , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Dysfunction of the intestinal barrier, as evidenced by increased intestinal permeability and bacterial translocation, has been reported under total parenteral nutrition (TPN). However, the role of Peyer's patches on the intestinal barrier in TPN is not well understood. We investigated whether TPN alters the uptake of microparticles by the follicle-associated epithelium of Peyer's patches. METHODS: Twenty rats were divided into two groups, a control group and a TPN group. Fluorescent polystyrene latex beads, 3.2 +/- 0.2 microns in diameter, were used as a probe for measuring the uptake by Peyer's patches. After 1 week of consuming either the control or TPN diet, rats were killed. On the day of killing, 0.1 mL of latex beads solution was injected into a 1-cm length of ileal loop, within 10 cm of the ileocecal valve. Samples were taken after 30 minutes of injection, sectioned by cryostat, and then viewed under a fluorescent microscope. Follicle-associated epithelial length and particles were counted using a confocal laser scanning microscope. The number of particles within each compartment was standardized per unit length of epithelium of Peyer's patches. RESULTS: Particle numbers within Peyer's patch dome of the TPN group were significantly increased compared with those of the control group (p < .01). CONCLUSIONS: These data suggest that dysfunction of the intestinal barrier in TPN might be associated with a change of uptake by Peyer's patches.
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Nutrición Parenteral Total , Ganglios Linfáticos Agregados/metabolismo , Animales , Masculino , Microesferas , Ratas , Ratas Sprague-DawleyRESUMEN
The purpose of this study was to investigate the intestinal hemodynamics and gut glutamine metabolism during endotoxemia, and their correlation with altered intestinal absorptive capacity and permeability. Seventeen Sprague-Dawley rats were used in the study. The endotoxin group (ENDO) received endotoxin (10 mg/kg intraperitoneally, n = 9), while the control group (CONT, n = 8) received saline injection. Twelve hours later, D-xylose (0.5 g/kg) and fluorescein isothiocyanate-dextran (FITC-dextran, 750 mg/kg) were given by oral gavage. One hour later abdominal aortic (AA) blood flow, superior mesenteric venous (SMV) flow, mean arterial pressure (MAP), central venous pressure (CVP), and SMV pressure (SMVP) were also measured. The MAP, AA, and SMV blood flow decreased (P < 0.05), while the CVP and SMVP increased (P < 0.05) in the ENDO group as compared with the CONT group. The ENDO group showed significant decreases for both intestinal glutaminase activity and net intestinal glutamine uptake (P < 0.05). The D-xylose concentration in SMV decreased significantly (P < 0.05) in the ENDO group as compared with the CONT group. However, the plasma FITC-dextran concentration showed no significant difference between the groups. Endotoxin produced a hypodynamic effect in rats 12 h after intraperitoneal administration in association with both a decreased intestinal glutamine metabolism and an absorptive capacity.
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Endotoxemia/fisiopatología , Glutamina/metabolismo , Intestinos/fisiología , Animales , Permeabilidad de la Membrana Celular , Técnica de Dilución de Colorante , Tránsito Gastrointestinal , Glutaminasa/metabolismo , Hemodinámica , Absorción Intestinal/fisiología , Intestinos/irrigación sanguínea , Intestinos/enzimología , Masculino , Ratas , Ratas Sprague-Dawley , Flujo Sanguíneo RegionalRESUMEN
We describe a case of successful laparoscopic resection of a left adrenal neuroblastoma (NB) detected by mass screening (MS) in an 8-month-old boy. Cases with MS NBs are supposed to be potential candidates for laparoscopic surgery in the pediatric age group.
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Neoplasias de las Glándulas Suprarrenales/cirugía , Laparoscopía , Neuroblastoma/cirugía , Humanos , Lactante , Laparoscopía/métodos , Masculino , Tamizaje Masivo , PuncionesRESUMEN
The number of elderly patients receiving home parenteral nutrition (HPN) has been increasing. This could result from several factors such as advances in HPN therapy and the natural aging of long-term older patients on HPN. Our experience shows that elderly patients receiving HPN are likely to be highly dependent on family members to supervise their HPN therapy. Therefore, indepth education of family members is essential before HPN is initiated. The important points to manage the elderly HPN patients are as follows: 1) It takes a long time for the elderly to learn the technique. 2) They often make technical mistakes. 3) In many cases, patients are not living with their children. Therefore, it is often difficult for them to support the patient's HPN therapy. 4) Nurses play an important role in giving instructions on catheter care and HPN techniques to the patient or family members.
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Nutrición Parenteral en el Domicilio/normas , Cuidado Terminal , Anciano , Anciano de 80 o más Años , Cuidadores , Femenino , Humanos , MasculinoRESUMEN
The intraluminal mucous gel layer across the small intestine of rats aged 3 days, 1, 2, 4 and 6 weeks (n=10x5) was studied to investigate its postnatal development. Celloidin stabilisation of dried cryostat sections of small intestine, with the luminal contents, preserved the intraluminal mucous gel layer for staining by the periodic acid-Schiff reaction. Morphological differences in the mucous gel, between the villi of the small intestine, in rats of several postnatal ages were observed, most notably after the age of 2 weeks. The adhesive mucous gel layer, covering the intestinal epithelium in the small intestine, appears to undergo rapid development after weaning.
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BACKGROUND: From the developmental aspects, the distribution of fluorescein isothiocyanate dextran 70,000 (FTTC-dextran) and mucous gel across the lumen of small intestine was observed as an investigation into the role of mucous gel on intestinal permeability. Furthermore, the effect of N-acetyl cysteine (NAC), a mucolytic agent, on intestinal permeability was examined. METHODS: In suckling and weaned rats, FTTC-dextran (750 mg/kg body wt) was gavage-fed. After 3 hours, blood samples were taken by cardiac puncture to analyze plasma FTTC-dextran by fluorescence spectrometry. Samples of small intestine with luminal contents were frozen and sectioned in a cryostat for fluorescence microscopy; the same sections were placed in a 0.2% celloidin solution to preserve mucous gel and were stained by periodic acid-Schiff reaction for light microscopy. In weaned rats, intestinal permeability was examined with different concentrations of intraluminally instilled NAC. RESULTS: The plasma level of FTTC-dextran showed a significant increase (p < .01) in suckling rats compared with the weaned rats. Morphologic findings were similar in both the jejunum and ileum: The spaces between villi were not entirely filled with mucus but filled with FTTC-dextran in suckling rats, whereas the spaces were filled with mucus and not filled with FTTC-dextran in weaned rats. Intestinal permeability in groups with NAC were significantly higher (p < .01) than that in group without NAC. CONCLUSIONS: These results suggest that an increase in the mucous gel layer that coats the epithelial lining according to the maturation of the gastrointestinal tract is one of the most important factors for a restriction in intestinal permeability.
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Mucosa Intestinal/crecimiento & desarrollo , Intestino Delgado/crecimiento & desarrollo , Moco/metabolismo , Acetilcisteína/farmacología , Animales , Dextranos/metabolismo , Epitelio/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Geles , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Intestino Delgado/efectos de los fármacos , Intestino Delgado/metabolismo , Masculino , Permeabilidad , Ratas , Ratas Sprague-Dawley , Espectrometría de FluorescenciaRESUMEN
OBJECTIVE: The authors determined the effects of alanyl-glutamine-supplemented total parenteral nutrition (TPN) on mucosal metabolism, integrity, and permeability of the small intestine in rats. METHODS: Male Sprague-Dawley rats were randomized to receive TPN supplemented with a conventional amino acids mixture (STD group) or the same solution supplemented with alanyl-glutamine; both solutions were isocaloric and isonitrogenous. On the seventh day of TPN, D-xylose and fluorescein isothiocyanate (FITC)-dextran were administered orally. One hour later, superior mesenteric vein (SMV) D-xylose and plasma FITC-dextran concentration were measured. Intestinal blood flow and calculated intestinal substrates flux were measured with ultrasonic transit time flowmetery. RESULTS: Plasma FITC-dextran increased significantly in the STD group. Intestinal blood flow and SMV D-xylose concentration did not differ between the groups. Mucosa weight, villus height, mucosal wall thickness, mucosal protein, and DNA and RNA content in jejunal mucosa were significantly increased in the alanyl-glutamine group. Jejunal mucosal glutaminase activity and net intestinal uptake of glutamine (glutamine flux) were significantly higher in the alanyl-glutamine group as compared with the STD group. CONCLUSION: Addition of alanyl-glutamine dipeptide to the TPN solution improves intestinal glutamine metabolism and prevents mucosal atrophy and deterioration of permeability.
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Dipéptidos/uso terapéutico , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/metabolismo , Nutrición Parenteral Total/métodos , Animales , Velocidad del Flujo Sanguíneo , Permeabilidad de la Membrana Celular/efectos de los fármacos , Dipéptidos/farmacología , Evaluación Preclínica de Medicamentos , Tránsito Gastrointestinal/efectos de los fármacos , Absorción Intestinal/efectos de los fármacos , Mucosa Intestinal/irrigación sanguínea , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
BACKGROUND: Although it has been reported that total parenteral nutrition induces an increased intestinal permeability and a decreased mucous gel layer covering the intestinal epithelium, the role of mucous gel on intestinal permeability has not been well understood. We examined the in vivo effects of N-acetyl cysteine (NAC) as mucolytic agent and colchicine as suppressant of the mucus production on the intestinal transmission of fluorescein isothiocyanate dextran 70,000 (FITC-dextran). METHODS: Rats were divided into four groups. In each group, FITC-dextran (750 mg/kg) with or without NAC (3000 mg/kg) was injected into the small intestinal lumen 3 hours after intraperitoneal injection of saline or colchicine (Col, 10 mg/kg). Thirty minutes after injection of FITC-dextran, blood samples were taken from portal vein to analyze plasma fluorescein concentration by fluorescence spectrometry. Samples of small intestine were sectioned in a cryostat for fluorescence microscopy, and the identical sections were stained by periodic acid-Schiff reaction. RESULTS: Plasma FITC-dextran level in NAC group was higher than that in control group (p < .01), that in Col + NAC group was higher than that in Col group (p < .01) and that in Col + NAC group was higher than that in NAC group (p < .05). The spaces between villi were filled with mucous gel in the control and Col groups, whereas those were not entirely filled with mucous gel in NAC and Col + NAC groups. FITC-dextran and mucous gel showed complementary distribution in all rats. The villous interstitial edema was recognized in NAC group and the villi were disrupted in Col + NAC group. CONCLUSIONS: These results suggest that intestinal permeability is possibly affected not only by the mucous gel covering the intestinal epithelium but also by mucus release from goblet cells of the small intestine.
Asunto(s)
Acetilcisteína/farmacología , Colchicina/farmacología , Mucosa Intestinal/fisiología , Moco/metabolismo , Adhesividad , Animales , Permeabilidad de la Membrana Celular/efectos de los fármacos , Dextranos/metabolismo , Fluoresceína-5-Isotiocianato/análogos & derivados , Fluoresceína-5-Isotiocianato/metabolismo , Geles , Mucosa Intestinal/efectos de los fármacos , Intestino Delgado/anatomía & histología , Intestino Delgado/fisiología , Masculino , Microscopía Fluorescente , Reacción del Ácido Peryódico de Schiff , Ratas , Ratas Sprague-DawleyRESUMEN
Glutamine (Gln)-supplemented total parenteral nutrition (TPN) has been shown to improve mucosal adaptation after massive small bowel resection (SBR); however, its influences on intestinal amino acid metabolism remain unknown. In this study, intestinal amino acid flux, circulating plasma aminogram, mucosal glutaminase activity and protein, and DNA content were measured 7 days after massive SBR in rats receiving either standard (Std) or Gln-supplemented TPN. Sham-operated rats and rats fed chow after enterectomy served as controls. The uptake of Gln and the release of citrulline (Cit) by the remaining intestine was significantly decreased, with reduced mucosal glutaminase activity after SBR in the Chow and Std-TPN groups. Glutamine supplementation resulted in significantly increased gut Gln uptake compared with Std-TPN (P < 0.01). Mucosal glutaminase activity, mucosal protein, and DNA content was also increased by Gln; however, the gut release of Cit remained unchanged (P > 0.05). The subsequent decrease in circulating arginine (Arg) in the Gln-TPN group compared with the Std-TPN group (P < 0.05) was attributed to an insufficient exogenous supply. These findings show that Gln-supplemented TPN improves mucosal growth and gut Gln uptake after SBR. However, the intestinal production of Cit, which remained low in both TPN groups, may lead to an insufficiency of endogenous Arg synthesis. Thus, both Gln and Arg may be essential amino acids after SBR.
Asunto(s)
Aminoácidos/metabolismo , Glutamina/administración & dosificación , Intestino Delgado/metabolismo , Intestino Delgado/cirugía , Nutrición Parenteral Total , Síndrome del Intestino Corto/terapia , Animales , Arginina/fisiología , Citrulina/metabolismo , Glutamina/fisiología , Mucosa Intestinal/metabolismo , Masculino , Ratas , Ratas Sprague-Dawley , Síndrome del Intestino Corto/metabolismoRESUMEN
The distribution of fluorescein isothiocyanate dextran 70,000 (FITC-dextran) and mucous gel across the lumen of small intestine was observed as an investigation into the role of mucous gel on permeability in total parenteral nutrition (TPN). Thirty-two rats were randomly divided into two groups fed with either TPN or oral rat food. On day 4 or 7, FITC-dextran (750 mg/kg body weight) was given through the gastroduodenal tube. After 1 hour, blood samples were taken by aortic puncture to analyze plasma FITC-dextran by fluorescence spectrometry. Samples of small intestine with luminal contents were frozen and sectioned in a cryostat for fluorescence microscopy; the same sections were placed in a 0.2% celloidin solution for 3 minutes to preserve mucous gel and stained by periodic acid-Schiff reaction for light microscopy. The plasma level of FITC-dextran after 1 hour of this marker injection showed a significant increase (p < .01) in the TPN group compared with the rat food group on days 4 and 7. Morphologic findings on days 4 and 7 were similar in both the jejunum and ileum: The mucous gel filled the spaces between villi and FITC-dextran centered in the lumen in the rat food group, whereas the mucous gel decreased and FITC-dextran filled the spaces between villi in the TPN group. FITC-dextran and mucous gel showed complementary distributions in both groups. These data suggest that TPN decreases luminal mucous gel and increases permeability of small intestine in rats.