RESUMO
BACKGROUND/AIMS: Previous studies from our laboratory have revealed impaired intestinal absorption of D-galactose in lipopolysaccharide-treated rabbits. The aim of the present work was to examine the effect of LPS on D-galactose intestinal absorption in vitro. METHODS: D-galactose intestinal transport was assessed employing three techniques: sugar uptake in rings of everted jejunum, transepithelial flux in Ussing-type chambers and transport assays in brush border membrane vesicles. The level of expression of the Na(+)/D-galactose cotransporter (SGLT1) was analyzed by Western blot. RESULTS: LPS decreased the mucosal D-galactose transport in rabbit jejunum but a preexposition to the endotoxin was required. LPS affected the Na(+)-dependent transport system by increasing the apparent Km value without affecting the Vmax. It also decreased the Na(+), K(+)-ATPase activity. However, it did not inhibit neither the uptake of D-galactose by brush border membrane vesicles nor modified the SGLT1 protein levels in the brush border, suggesting an indirect endotoxin effect. This inhibitory effect, was reduced by selective inhibitors of Ca(2+)-calmodulin (W13), protein kinase C (GF 109203X), p38 mitogen-activated protein kinase (SB 203580), c-Jun N-terminal kinase (SP 600125) and mitogen extracellular kinase (U 0126). CONCLUSION: LPS inhibits the mucosal Na(+)-dependent D-galactose intestinal absorption and the Na(+), K(+)-ATPase activity when it is added to the tissue. Intracellular processes related to protein kinases seem to be implicated in the endotoxin effect.
Assuntos
Galactose/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/efeitos dos fármacos , Lipopolissacarídeos/farmacologia , Animais , Transporte Biológico/efeitos dos fármacos , Cálcio/metabolismo , Calmodulina/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Relação Dose-Resposta a Droga , Técnicas In Vitro , Absorção Intestinal/efeitos dos fármacos , Espaço Intracelular/efeitos dos fármacos , Espaço Intracelular/metabolismo , Jejuno/efeitos dos fármacos , Jejuno/enzimologia , Cinética , Proteína Quinase C/metabolismo , Coelhos , Transportador 1 de Glucose-Sódio/metabolismoRESUMO
A case of myositis ossificans circumscripta in an eight-year-old, male, Weimaraner dog, is described. The animal was originally submitted with presumptive diagnosis of hernia. The lesion was a firm, subcutaneous nodule, 5 cm long by 3 cm width, localized in the right perineum muscles. After the radiologic study the lesion was interpreted as a neoplasm from coccygeal vertebrae. The surgical procedure to eliminate the neoplastic tissue confirmed its hardness. Excision was not complete and the sample was submitted to pathology. The mass was hard, well circumscribed, vascularized and surrounded by muscle tissue. The histopathological study revealed a heterotopic ossification with zones of atrophy, degeneration and muscle necrosis, alternating with zones of trabecular ossification toward the periphery, limited by abundant fibrous connective tissue. These findings are characteristic of myositis ossificans circumscripta. This case represents the first report in a dog in Mexico, of a condition resembling the myositis ossificans circumscripta in humans.
Se describe un caso de miositis osificante localizada en un perro de raza Weimaraner, macho, de ocho años de edad. El animal fue originalmente remitido con diagnóstico presuntivo de hernia. La lesión era un nódulo subcutáneo firme con 5 cm de largo por 3 cm de ancho, localizada en los músculos del perineo derecho. Después del estudio radiológico la lesión fue interpretada como una neoplasia originada de vértebras coccígeas. Su dureza se constató durante el procedimiento quirúrgico para eliminar la neoformación. La lesión se extirpó parcialmente y la muestra se remitió a patología. La masa era dura, bien delimitada, vascularizada y con tejido muscular a su alrededor. El estudio histopatológico reveló osificación heterotópica con zonas de atrofia, degeneración y necrosis muscular, alternando con zonas de osificación trabecular en la periferia, limitadas por abundante tejido conectivo fibroso. Estos hallazgos son característicos de la miositis osificante localizada. Este caso constituye el primer informe de esta condición patológica en un perro en México, similar a la miositis osificante localizada en humanos.
RESUMO
Lipopolysaccharide (LPS) endotoxin is a causative agent of sepsis. The aim of this study was to examine LPS effects on intestinal fructose absorption and to decipher mechanisms. Sepsis was induced by intravenous injection of LPS in rabbits. The ultrastructural study and DNA fragmentation patterns were identical in the intestine of LPS and sham animals. LPS treatment reduced fructose absorption altering both mucosal-to-serosal transepithelial fluxes and uptake into brush border membrane vesicles (BBMVs). Cytochalasin B was ineffective on fructose uptake, indicating that GLUT5, but not GLUT2, transport activity was targeted. GLUT5 protein levels in BBMvs were lower in LPS than in sham-injected rabbits. Thus lower fructose transport resulted from lower levels of GLUT5 protein. LPS treatment decreased GLUT5 levels by proteasome-dependent degradation. Specific inhibitors of PKC, PKA, and MAP kinases (p38MAPK, JNK, MEK1/2) protected fructose uptake from adverse LPS effect. Moreover, a TNF-alpha antagonist blocked LPS action on fructose uptake. We conclude that intestinal fructose transport inhibition by LPS is associated with diminished GLUT5 numbers in the brush border membrane of enterocytes triggered by activation of several interrelated signaling cascades and proteasome degradation.