RESUMO
Inflammatory bowel diseases (IBD) are chronic and relapsing inflammatory conditions of the gastrointestinal tract including Crohn's disease (CD) and ulcerative colitis (UC). Galectins, defined by shared consensus amino acid sequence and affinity for ß-galactosides, are critical modulators of the inflammatory response. However, the relevance of the galectin network in the pathogenesis of human IBD has not yet been explored. Here, we analyzed the expression of relevant members of the galectin family in intestinal biopsies, and identified their contribution as novel mucosal markers in IBD. Colonic biopsies were obtained from 59 IBD patients (22 CD and 37 UC), 9 patients with gut rejection after transplantation, 8 adult celiac patients, and 32 non-IBD donors. Galectin mRNA expression was analyzed by RT-PCR and qPCR using specific primers for individual galectins. A linear discriminant analysis (LDA) was used to analyze galectin expression in individual intestinal samples. Expression of common mucosal-associated galectins (Gal-1, -3, -4, -9) is dysregulated in inflamed tissues of IBD patients compared with non-inflamed IBD or control samples. LDA discriminated between different inflammation grades in active IBD and showed that remission IBD samples were clusterized with control samples. Galectin profiling could not distinguish CD and UC. Furthermore, inflamed IBD was discriminated from inflamed tissue of rejected gut in transplanted patients and duodenum of celiac patients, which could not be distinguished from control duodenum samples. The integrative analysis of galectins discriminated IBD from other intestinal inflammatory conditions and could be used as potential mucosal biomarker.
Assuntos
Colite Ulcerativa/genética , Doença de Crohn/genética , Galectina 3/biossíntese , Galectina 4/biossíntese , Galectinas/biossíntese , Inflamação/genética , Tirosina/análogos & derivados , Adulto , Benzamidas , Biópsia , Colite Ulcerativa/patologia , Doença de Crohn/patologia , Feminino , Galectina 3/genética , Galectina 4/genética , Galectinas/genética , Trato Gastrointestinal/metabolismo , Trato Gastrointestinal/patologia , Regulação da Expressão Gênica , Humanos , Inflamação/patologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Intestinos/patologia , Masculino , RNA Mensageiro/biossíntese , Tirosina/biossíntese , Tirosina/genéticaRESUMO
BACKGROUND: Fabry disease is an X-linked disorder that results from the deficiency of the lysosomal enzyme alpha-galactosidase A. The defect leads to the accumulation of globotriaosylceramide (Gb3). The detection of Gb3 accumulated in different tissues may help in the diagnosis and enzyme replacement therapy monitoring. For this reason, we developed a simple method available to clinical laboratories to measure this analyte. METHODS: Gb3 excretion was determined by the incubation of urine sediment glycolipids from Fabry patients with agalsidase alpha and subsequent determination of galactose produced. RESULTS: The amount of urinary Gb3 in Fabry hemizygotes was significantly higher (p = 0.00001) than the amount in normal controls. Patients undergoing enzyme replacement therapy with agalsidase alpha showed a significantly lower content of Gb3 in urine sediment. This method showed a good recovery and comparability with a previously validated method. CONCLUSIONS: We developed an easy method for quantification of Gb3 in urine samples from Fabry patients, by the use of the specific recombinant enzyme for this glycolipid, that does not require complex infrastructure. Urinary Gb3 as measured by this enzymatic method could be useful for the diagnosis and monitoring of treatment in Fabry patients.
