RESUMO
BACKGROUND: Obesity is a risk factor for colorectal cancer, yet metabolic distinctions between healthy right and left colon tissue, before cancer is diagnosed, remains largely unknown. This study compared right-ascending and left-descending colon tissue metabolomes to identify differences from the stool metabolome in normal weight, overweight, and obese adults. AIM: To examine right and left colon tissue metabolites according to body mass index that may serve as mechanistic targets for interventions and biomarkers for colon cancer risk. METHODS: Global, non-targeted metabolomics was applied to assess right-ascending and left-descending colon tissue collected from healthy adults undergoing screening colonoscopies to test the hypothesis that BMI differentially impacts colon tissue metabolite profiles. The colon tissue and stool metabolome of healthy adults (n = 24) was analyzed for metabolite signatures and metabolic pathway networks implicated in progression of colorectal cancer. RESULTS: Ascending and descending colon contained 504 host, food, and microbiota-derived metabolites from normal weight, overweight and obese adults grouped according to body mass index. Amino acids, lipids, and nucleotides were among the chemical types that further differentiated from the stool metabolite profiles. Normal weight adults had 46 significantly different metabolites between ascending and descending colon tissue locations, whereas there were 37 metabolite differences in overweight and 28 metabolite differences for obese adults (P < 0.05). Obese adults had trimethylamine N-oxide, endocannabinoids and monoacylglycerols with different relative abundances identified between ascending and descending colon. Primary and secondary bile acids, vitamins, and fatty acids also showed marked relative abundance differences in colon tissue from overweight/obese adults. CONCLUSION: There were metabolite profile differences between right-ascending and left-descending colon tissue in healthy adults. Colon lipids and other metabolites in obese and overweight adults were distinguished from normal weight participants and associated with gut inflammation, nutrient absorption, and products of microbiota metabolism.
Assuntos
Colo Ascendente/metabolismo , Colo Descendente/metabolismo , Metaboloma , Obesidade/metabolismo , Sobrepeso/metabolismo , Adulto , Biomarcadores Tumorais/metabolismo , Índice de Massa Corporal , Neoplasias Colorretais/etiologia , Fezes/química , Feminino , Microbioma Gastrointestinal/fisiologia , Voluntários Saudáveis , Humanos , Peso Corporal Ideal/fisiologia , Absorção Intestinal/fisiologia , Metabolismo dos Lipídeos , Lipídeos/análise , Masculino , Pessoa de Meia-Idade , Obesidade/complicações , Sobrepeso/complicações , Fatores de RiscoRESUMO
Cytochrome P450 (CYP) 3A4 and P-glycoprotein (P-gp) have broad substrate overlap and are involved in the metabolism and transport of nearly 50% of currently prescribed medications. In the intestine, CYP3A4 and P-gp are coexpressed in the enterocytes at the intestinal villous tip and act in a coordinated manner to limit drug and xenobiotic oral bioavailability prior to further metabolism and disposition in the liver. Crohn's disease (CD), a form of inflammatory bowel disease, introduces a transmural intestinal insult that disrupts the intestinal barrier function; it therefore has the potential to affect intestinal drug metabolism and transport. We hypothesized that individuals with CD have reduced intestinal expression of CYP3A4 and P-gp. We obtained intestinal biopsy samples from individuals with and without CD and quantified the expression of CYP3A4 and P-gp. When we carried out Western analysis for protein expression, we observed a significant reduction in ileal (45% decrease) and colonic (78% decrease) CYP3A4 protein expression in subjects with CD compared with those without. Similarly, an 85% reduction in colonic P-gp protein expression was seen in the CD patients. Our data highlight important and novel findings pertaining to CD-associated changes to the intestinal expression of CYP3A4 and P-gp that are of relevance to better predict substrate drug dosing for patients with CD.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Doença de Crohn/metabolismo , Citocromo P-450 CYP3A/metabolismo , Mucosa Intestinal/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Biópsia , Células CACO-2 , Colo Ascendente/metabolismo , Colo Ascendente/patologia , Doença de Crohn/patologia , Enterócitos/metabolismo , Feminino , Humanos , Íleo/metabolismo , Íleo/patologia , Masculino , Proteínas dos Microfilamentos/metabolismo , Pessoa de Meia-Idade , Adulto JovemRESUMO
Developing rapidly from the cecal diverticulum in a 5-week-old embryo, the cecum, which is developed from the caudal limb of the midgut loop, is different from the ascending colon. The aim of this study was to analyze the different clinicopathological and biological characteristics of patients with carcinoma of the cecum and ascending colon. We accessed data for 59,035 patients with adenocarcinomas of the cecum and ascending colon from the Surveillance, Epidemiology and End Results database to explore the potential associations between the clinicopathological characteristics and overall survival. Furthermore, we analyzed the differences in gene expression between the two segments in the Gene Expression Omnibus database. The results were validated in The Cancer Genome Atlas database, as well as with another independent dataset from the First Affiliated Hospital of Xi'an Jiaotong University. The results of this study revealed the potential prognostic differences between adenocarcinoma of the cecum and ascending colon, which may be caused by the differential expression levels of the SLCO1B3 gene. When including the expression levels of SLCO1B3 in intraoperatively examined lymph nodes, 8 factors were found able to predict the prognosis of patients with carcinomas of the cecum and ascending colon. As regards the surgical therapeutic strategies, the resection of >15 local lymph nodes is appropriate for improving the prognosis of patients.
Assuntos
Carcinoma/genética , Ceco/metabolismo , Colo Ascendente/metabolismo , Membro 1B3 da Família de Transportadores de Ânion Orgânico Carreador de Soluto/genética , Adulto , Idoso , Carcinoma/patologia , Ceco/patologia , Colo Ascendente/embriologia , Colo Ascendente/patologia , Intervalo Livre de Doença , Feminino , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Estimativa de Kaplan-Meier , Linfonodos/metabolismo , Linfonodos/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteoma/genéticaRESUMO
BACKGROUND: Survivin, a member of the inhibitor of apoptosis protein (IAP) family, regulates mitosis and chromosome segregation. The expression of survivin proceeds during embryonic development and in addition has already been demonstrated in cancer cells. However, there is also evidence of survivin expression in differentiated tissues, including the gastro-intestinal tract of adult rats. A study with human colon specimens exhibited survivin in most basal crypt epithelial cells of normal mucosa. There is rather limited information on survivin expression in the small intestine. In order to paint a more detailed and thus complete picture of survivin expression patterns in the gastrointestinal tract, we used an immunohistochemical approach in normal adult rat small intestinal and ascending colonic tissue. Moreover, to get deeper insights in the regulation of survivin expression after tissue damage, we also studied its expression in mesenteric ischemia-reperfusion (I/R) injury. METHODS: Mesenteric ischemia-reperfusion injury was induced in male Wistar rats (six animals/group) by occlusion of the superior mesenteric artery for 90 min and subsequent reperfusion for 120 min. Paraffin sections of untreated or ischemically treated tissue were assessed immunohistochemically by survivin and Ki-67 staining. RESULTS: Survivin could be detected in the small intestine and ascending colon of the normoxia group. It was expressed mainly in the epithelial cells of the crypts and only marginally in the villi. The individual small intestinal segments studied revealed comparable staining intensities. Likewise, expression of survivin was detected in the ischemically damaged small intestine and ascending colon. The expression pattern corresponded to the normoxic animals, as far as verifiable due to the existing tissue damage. Comparison of the expression pattern of Ki-67, a protein that acts as a cellular marker for proliferation, and survivin demonstrated a coincidental localization of the two proteins in the small intestinal and ascending colonic tissue. CONCLUSIONS: Survivin was expressed strongly in epithelial cells of small intestinal as well as ascending colonic tissue. Its expression was located in cells with a high proliferation rate and regenerative capacity. This further supports the decisive role of survivin in cell division. Surprisingly, the ischemically damaged small intestinal and ascending colonic tissue showed a comparably high expression level. These results suggest that there is already a maximal survivin expression under normal conditions. However, the intestine is able to maintain the regenerative capacity even in spite of an ischemic injury. These findings reflect the important relevance of an intact intestinal barrier.
Assuntos
Colo Ascendente/metabolismo , Intestino Delgado/metabolismo , Isquemia Mesentérica/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Traumatismo por Reperfusão/metabolismo , Animais , Colo Ascendente/irrigação sanguínea , Intestino Delgado/irrigação sanguínea , Masculino , Ratos , Ratos Wistar , SurvivinaRESUMO
BACKGROUND AND AIM: The microRNA (miRNA) expression profiles of the terminal ileum, sigmoid colon, and rectal mucosa of adult patients with active Crohn's disease (CD) have been previously reported. The purpose of this study was to identify dysregulated miRNAs in the mucosa of the ascending colon. METHODS: Biopsy tissue samples were taken from the mucosae of inflammatory (iCD) or noninflammatory (niCD) areas of the ascending colons of adult patients with active CD. miRNA and mRNA expression profiles were detected using microarray analyses. miRNAs and messenger RNAs (mRNAs) demonstrating significant differences were validated via quantitative real-time polymerase chain reaction. Luciferase reporter genes were used to measure two miRNAs inhibition of potential target genes in human 293T cells in vitro. RESULTS: Compared with the healthy control group, the ascending colon miRNA expression profiles revealed that 43 miRNAs were significantly upregulated and 35 were downregulated in the iCD group. The mRNA expression profiles indicated that 3370 transcripts were significantly differentially expressed in the ascending colon, with 2169 upregulated and 1201 downregulated mRNAs in the iCD group, and only 20 miRNAs demonstrated significant differential expression in the niCD group. In contrast, nearly 100 miRNAs significantly varied between the iCD and niCD groups. Finally, luciferase reporter gene assays showed that hsa-miR-16-1 directly regulated the human C10orf54 gene and that they were negatively correlated. CONCLUSIONS: Our results indicated that the differentially expressed miRNAs and mRNAs were related to immune inflammation and intestinal flora. The data provide preliminary evidence that the occurrence of CD involves the inhibition of C10orf54 expression by hsa-miR-16-1.
Assuntos
Colo Ascendente/metabolismo , Doença de Crohn/genética , Doença de Crohn/metabolismo , Mucosa Intestinal/metabolismo , MicroRNAs/genética , Transcriptoma/genética , Colo Ascendente/microbiologia , Microbioma Gastrointestinal , Células HEK293 , Humanos , Inflamação/genética , Inflamação/metabolismo , Mucosa Intestinal/microbiologia , MicroRNAs/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas Supressoras de TumorRESUMO
OBJECTIVE: Accurate differentiation between Crohn's disease (CD) and UC is important to ensure early and appropriate therapeutic intervention. We sought to identify proteins that enable differentiation between CD and UC in children with new onset IBD. DESIGN: Mucosal biopsies were obtained from children undergoing baseline diagnostic endoscopy prior to therapeutic interventions. Using a super-stable isotope labeling with amino acids in cell culture (SILAC)-based approach, the proteomes of 99 paediatric control and biopsies of patients with CD and UC were compared. Multivariate analysis of a subset of these (n=50) was applied to identify novel biomarkers, which were validated in a second subset (n=49). RESULTS: In the discovery cohort, a panel of five proteins was sufficient to distinguish control from IBD-affected tissue biopsies with an AUC of 1.0 (95% CI 0.99 to 1.0); a second panel of 12 proteins segregated inflamed CD from UC within an AUC of 0.95 (95% CI 0.86 to 1.0). Application of the two panels to the validation cohort resulted in accurate classification of 95.9% (IBD from control) and 80% (CD from UC) of patients. 116 proteins were identified to have correlation with the severity of disease, four of which were components of the two panels, including visfatin and metallothionein-2. CONCLUSIONS: This study has identified two panels of candidate biomarkers for the diagnosis of IBD and the differentiation of IBD subtypes to guide appropriate therapeutic interventions in paediatric patients.
Assuntos
Colite Ulcerativa , Colo Ascendente , Doença de Crohn , Subunidade beta da Proteína Mitocondrial Trifuncional/análise , Nicotinamida Fosforribosiltransferase/análise , Proteômica/métodos , Adolescente , Biomarcadores/análise , Biópsia/métodos , Canadá , Colite Ulcerativa/diagnóstico , Colite Ulcerativa/metabolismo , Colite Ulcerativa/patologia , Colo Ascendente/metabolismo , Colo Ascendente/patologia , Doença de Crohn/diagnóstico , Doença de Crohn/metabolismo , Doença de Crohn/patologia , Estudos Transversais , Diagnóstico Diferencial , Intervenção Médica Precoce , Feminino , Humanos , Masculino , Seleção de PacientesRESUMO
BACKGROUND: Colorectal cancer (CRC) is the most common form of cancer and the third leading cause of death in Taiwan. Serum alpha-fetoprotein (AFP) has been extensively used as a biomarker for hepatocellular carcinoma (HCC) and yolk sac tumors. CASE PRESENTATION: This case report presents a 90-year-old woman with right abdominal pain and poor appetite for 1 week. The computed tomography (CT) showed wall thickening in the proximal ascending colon with ruptured appendicitis. Preoperative serum AFP was high. There was no definite liver metastasis or other abnormal findings in the hepatobiliary systems. After initial empirical antibiotic treatment, we performed laparoscopic right hemicolectomy. The pathological assessment was poorly differentiated adenocarcinoma with neuroendocrine differentiation in the ascending colon. The tumor cells did not produce AFP. Amazingly, the follow-up serum AFP level 1 month after the surgery declined to normal range. The patient had an uneventful course after the surgery and was free of recurrence or metastasis within 5 months of follow-up. CONCLUSIONS: AFP may be a useful tumor marker in poorly differentiated colorectal cancer with neuroendocrine component patients and a prediction of early treatment response.
Assuntos
Adenocarcinoma/patologia , Carcinoma Neuroendócrino/patologia , Diferenciação Celular , Colo Ascendente/patologia , Neoplasias do Colo/patologia , alfa-Fetoproteínas/análise , Adenocarcinoma/sangue , Adenocarcinoma/cirurgia , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/sangue , Carcinoma Neuroendócrino/sangue , Carcinoma Neuroendócrino/cirurgia , Colo Ascendente/metabolismo , Colo Ascendente/cirurgia , Neoplasias do Colo/sangue , Neoplasias do Colo/cirurgia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Prognóstico , Tomografia Computadorizada por Raios XRESUMO
AIM: The present study was designed to investigate the function and clinical implications of stool short-chain fatty acids (SCFAs) in patients with mixed refractory constipation. METHOD: Ascending colon specimens obtained from 30 patients with ascending colon cancer were regarded as the control group. Ascending colon specimens obtained from patients with mixed refractory constipation were regarded as the experimental group and were divided into three subgroups, according to Wexner scores [A constipation scoring system to simplify evaluation and management of constipated patients. Dis Colon Rectum 1996; 39: 681-5] of 16-20, 21-25 and 26-30, with 30 patients in each group. The stool SCFAs were extracted and quantitatively analysed using gas chromatography-mass spectrometry (GC-MS). The expression of G protein-coupled receptor 43 (GPR43) and of choline acetyltransferase (ChAT) were detected by immunofluorescence, reverse transcription-polymerase chain reaction (RT-PCR) and western blotting of colon samples. RESULTS: The levels of acetate, propionate and butyrate were significantly lower in the experimental group than in the control group (P < 0.05). Compared with the control group, the densitometric quantification and mean density of GPR43 and ChAT proteins, and expression of GPR43 and CHAT genes, were significantly decreased in the patients with mixed refractory constipation (P < 0.05). CONCLUSION: In the patients with mixed refractory constipation, the levels of stool SCFAs, including acetate, propionate and butyrate, as well as the levels of GPR43 and ChAT expressed in the colon, which were all negatively correlated with the Wexner score, were decreased and may be associated with the pathogenesis of mixed refractory constipation.
Assuntos
Colina O-Acetiltransferase/genética , Colo Ascendente/metabolismo , Constipação Intestinal/metabolismo , Ácidos Graxos Voláteis/metabolismo , Receptores de Superfície Celular/genética , Acetatos/metabolismo , Adolescente , Adulto , Western Blotting , Butiratos/metabolismo , Estudos de Casos e Controles , Colina O-Acetiltransferase/metabolismo , Neoplasias do Colo/genética , Neoplasias do Colo/metabolismo , Constipação Intestinal/genética , Fezes/química , Feminino , Imunofluorescência , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Masculino , Pessoa de Meia-Idade , Propionatos/metabolismo , Receptores de Superfície Celular/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto JovemRESUMO
G protein-coupled receptor 43/free fatty acid receptor 2 (GPR43/FFAR2) is essential for polymorphonuclear (PMN) recruitment. We investigated the expression of GPR43/FFAR2 in the colon from Crohn's disease patients and whether dietary fiber in enteral nutrition increases GPR43+ polymorphonuclear infiltration in mucosa. Segments of ascending colon and white blood cells from peripheral blood were obtained from 46 Crohn's disease patients and 10 colon cancer patients. The Crohn's disease patients were grouped by the activity of disease (active or remission) and enteral nutrition with or without dietary fiber. Histological feature, expression and location of GPR43/FFAR2 and level of tumor necrosis factor-α (TNF-α), interleukine-6 (IL-6) and myeloperoxidase were assessed. The results of hematoxylin-eosin and immunohistochemistry staining revealed that the infiltration of immune cells, including GPR43+ PMN, was more severe in active Crohn's disease patients who consumed normal food or enteral nutrition with dietary fiber than in remission patients and colon cancer patients. This finding was supported by the results of GPR43 and myeloperoxidase expression. Active Crohn's disease (CD) patients who consumed enteral nutrition without dietary fiber exhibited severe immune cell infiltration similar to the other active CD patients, but GPR43+ PMNs were rarely observed. The level of TNF-α mRNA in active Crohn's disease patients was higher than those of the other patients. In conclusion, the use of dietary fiber in enteral nutrition by active Crohn's disease patients might increase GPR43+ PMNs infiltration in colon mucosa. This effect was not observed in Crohn's disease patients in remission.
Assuntos
Doença de Crohn/imunologia , Fibras na Dieta/administração & dosagem , Mucosa Intestinal/imunologia , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Receptores de Superfície Celular/metabolismo , Adulto , Colo Ascendente/imunologia , Colo Ascendente/metabolismo , Neoplasias do Colo/sangue , Neoplasias do Colo/dietoterapia , Neoplasias do Colo/imunologia , Doença de Crohn/sangue , Doença de Crohn/dietoterapia , Nutrição Enteral/métodos , Feminino , Humanos , Interleucina-6/metabolismo , Mucosa Intestinal/metabolismo , Masculino , Pessoa de Meia-Idade , Peroxidase/metabolismo , Receptores de Superfície Celular/imunologia , Indução de Remissão , Estudos Retrospectivos , Fator de Necrose Tumoral alfa/metabolismoRESUMO
PURPOSE: Characterize the contents of distal ileum and cecum in healthy adults under conditions simulating the bioavailability/bioequivelance studies of drug products in fasted and fed state. METHODS: Twelve males participated in a two-phase crossover study. Phase I: subjects remained fasted overnight plus 4.5 h in the morning prior to colonoscopy. Phase II: subjects remained fasted overnight, consumed breakfast in the morning, and abstain from food until colonoscopy, 4.5 h after breakfast. Upon sampling, volume, pH and buffer capacity were measured; after ultracentrifugation, supernatant was physicochemically characterized and non-liquid particles diameter was measured. RESULTS: In distal ileum, pH is ~8.1 and size of non-liquid particles is ~200 µm, regardless of dosing conditions; in fed state, liquid fraction was lower whereas osmolality and carbohydrate content were higher. In cecum, the environment was similar with previously characterized environment in the ascending colon; in fasted state, size of non-liquid particles is smaller than in distal ileum (~70 µm). Fluid composition in distal ileum is different from cecum, especially in fasted state. CONCLUSION: Differences in luminal environment between distal ileum and cecum may impact the performance of orally administered products which deliver drug during residence in lower intestine. Dosing conditions affect cecal environment more than in distal ileum.
Assuntos
Ceco/metabolismo , Íleo/metabolismo , Preparações Farmacêuticas/metabolismo , Administração Oral , Adulto , Disponibilidade Biológica , Soluções Tampão , Colo Ascendente/metabolismo , Estudos Cross-Over , Jejum/metabolismo , Alimentos , Voluntários Saudáveis , Humanos , Concentração de Íons de Hidrogênio , Masculino , Concentração Osmolar , Equivalência Terapêutica , Adulto JovemRESUMO
BACKGROUND: Pannexin-2 (Panx2) is a member of the novel group of membrane spanning protein channels present in the central nervous system. Limited studies have examined Panx2 in the intestine, where it may have important physiological roles. The present study characterized Panx2 expression and localization in the human colon in health and disease states. METHODS: Immunofluorescence determined Panx2 localization and co-localization, and quantitative real-time PCR and Western blot determined gene and protein expression in ulcerative colitis (UC), Crohn's disease (CD), and control human colon. KEY RESULTS: Panx2 was widely expressed in myenteric and submucosal ganglia, particularly in the cytoplasm of neurons. Panx2 was also expressed on smooth muscle of the muscularis and blood vessels, some non-lymphoid leukocytes, mast cells, and mucosal epithelial cells. Co-localization of Panx2 occurred with ß-tubulin, neuronal nitric oxide synthase, substance P, vesicular acetylcholine transporter, and calcitonin gene-related peptide, indicating widespread Panx2 expression in extrinsic and intrinsic neurons. Molecular studies revealed a 3.4-fold higher level of Panx2 mRNA in ascending compared to sigmoid muscularis (p < 0.05), despite similar protein levels. Similarly, UC muscularis showed a 35-fold up-regulation in Panx2 mRNA, but not in protein (p < 0.05). CONCLUSIONS & INFERENCES: Here, we demonstrated the dense expression of Panx2 in the enteric nervous system and the co-localization of Panx2 with a spectrum of neuronal markers, indicating that Panx2 may be involved in mediating neurotransmission in the colon. The substantial increase in Panx2 mRNA in UC muscle but not protein suggests that the Panx2 translation process may be disrupted in UC.
Assuntos
Colite Ulcerativa/genética , Colo/metabolismo , Conexinas/genética , Doença de Crohn/genética , Sistema Nervoso Entérico/metabolismo , Neurônios/metabolismo , RNA Mensageiro/metabolismo , Adulto , Idoso , Western Blotting , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Estudos de Casos e Controles , Colite Ulcerativa/metabolismo , Colo Ascendente/metabolismo , Colo Sigmoide/metabolismo , Conexinas/metabolismo , Doença de Crohn/metabolismo , Células Epiteliais/metabolismo , Feminino , Humanos , Mucosa Intestinal/metabolismo , Leucócitos/metabolismo , Masculino , Mastócitos/metabolismo , Pessoa de Meia-Idade , Músculo Liso/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Substância P/metabolismo , Proteínas Vesiculares de Transporte de Acetilcolina/metabolismo , Adulto JovemRESUMO
Effective antiretroviral therapy (ART) dramatically reduces AIDS-related complications, yet the life expectancy of long-term ART-treated HIV-infected patients remains shortened compared to that of uninfected controls, due to increased risk of non-AIDS related morbidities. Many propose that these complications result from translocated microbial products from the gut that stimulate systemic inflammation--a consequence of increased intestinal paracellular permeability that persists in this population. Concurrent intestinal immunodeficiency and structural barrier deterioration are postulated to drive microbial translocation, and direct evidence of intestinal epithelial breakdown has been reported in untreated pathogenic SIV infection of rhesus macaques. To assess and characterize the extent of epithelial cell damage in virally-suppressed HIV-infected patients, we analyzed intestinal biopsy tissues for changes in the epithelium at the cellular and molecular level. The intestinal epithelium in the HIV gut is grossly intact, exhibiting no decreases in the relative abundance and packing of intestinal epithelial cells. We found no evidence for structural and subcellular localization changes in intestinal epithelial tight junctions (TJ), but observed significant decreases in the colonic, but not terminal ileal, transcript levels of TJ components in the HIV+ cohort. This result is confirmed by a reduction in TJ proteins in the descending colon of HIV+ patients. In the HIV+ cohort, colonic TJ transcript levels progressively decreased along the proximal-to-distal axis. In contrast, expression levels of the same TJ transcripts stayed unchanged, or progressively increased, from the proximal-to-distal gut in the healthy controls. Non-TJ intestinal epithelial cell-specific mRNAs reveal differing patterns of HIV-associated transcriptional alteration, arguing for an overall change in intestinal epithelial transcriptional regulation in the HIV colon. These findings suggest that persistent intestinal epithelial dysregulation involving a reduction in TJ expression is a mechanism driving increases in colonic permeability and microbial translocation in the ART-treated HIV-infected patient, and a possible immunopathogenic factor for non-AIDS related complications.
Assuntos
Fármacos Anti-HIV/efeitos adversos , Colo/efeitos dos fármacos , Regulação para Baixo/efeitos dos fármacos , Infecções por HIV/tratamento farmacológico , Mucosa Intestinal/efeitos dos fármacos , Proteínas de Junções Íntimas/antagonistas & inibidores , Junções Íntimas/efeitos dos fármacos , Centros Médicos Acadêmicos , Fármacos Anti-HIV/uso terapêutico , Estudos de Coortes , Colo/metabolismo , Colo/patologia , Colo/virologia , Colo Ascendente/efeitos dos fármacos , Colo Ascendente/metabolismo , Colo Ascendente/patologia , Colo Ascendente/virologia , Colo Descendente/efeitos dos fármacos , Colo Descendente/metabolismo , Colo Descendente/patologia , Colo Descendente/virologia , Colo Transverso/efeitos dos fármacos , Colo Transverso/metabolismo , Colo Transverso/patologia , Colo Transverso/virologia , Feminino , Infecções por HIV/metabolismo , Infecções por HIV/patologia , Infecções por HIV/virologia , Humanos , Íleo/efeitos dos fármacos , Íleo/metabolismo , Íleo/patologia , Íleo/virologia , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Mucosa Intestinal/virologia , Masculino , Pessoa de Meia-Idade , Ohio , Especificidade de Órgãos , Permeabilidade/efeitos dos fármacos , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Junções Íntimas/metabolismo , Junções Íntimas/patologia , Junções Íntimas/virologiaRESUMO
Lactoferrin (LF) is a multifunctional immune protein found at high concentrations in human milk. Herein, the effect of dietary bovine LF (bLF) on mucosal and systemic immune development was investigated. Colostrum-deprived piglets were fed formula containing 130 [control (Ctrl)], 367 (LF1), or 1300 (LF3) mg of bLF/(kg body weight · d). To provide passive immunity, sow serum was provided orally during the first 36 h of life. Blood, spleen, mesenteric lymph node (MLN), and ascending colon (Asc) contents were collected on day 7 (n = 10-14/group) and day 14 (n = 10-12/group). Immune cell populations were quantified by flow cytometry and immunoglobulins (Igs) were measured by ELISA. Additionally, immune cells were isolated from spleen and MLNs (n = 7/group) on day 7 and stimulated ex vivo with phytohemagglutinin or lipopolysaccharide (LPS) ± LF for 72 h. Secreted cytokine concentrations were quantified by multiplex assay. Lymphocyte populations [cluster determinant (CD)4, CD8, and natural killer cells] developed normally and were unaffected by dietary bLF. LF3 piglets tended to have 1.4 to 2 times more serum IgG than Ctrl piglets (P = 0.07) or LF1 piglets (P = 0.03), but IgA in Asc contents was unaffected by bLF. Asc IgA was 4 times higher on day 14 than day 7. Spleen cells from LF3 piglets produced 2 times more interleukin (IL)-10 and tumor necrosis factor (TNF)-α ex vivo than those from Ctrl or LF1 piglets. MLN cells from LF1 and LF3 piglets produced 40% more IL-10 and tended to produce 40% more IL-6 (P = 0.05) than those from Ctrl piglets. However, ex vivo bLF did not affect the cytokine response of spleen or MLN cells to LPS. In summary, dietary bLF alters the capacity of MLN and spleen immune cells to respond to stimulation, supporting a role for LF in the initiation of protective immune responses in these immunologically challenged neonates.
Assuntos
Imunidade Inata , Imunidade nas Mucosas , Imunomodulação , Fórmulas Infantis , Lactoferrina/metabolismo , Leucócitos Mononucleares/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Animais Recém-Nascidos , Bovinos , Células Cultivadas , Colo Ascendente/citologia , Colo Ascendente/crescimento & desenvolvimento , Colo Ascendente/imunologia , Colo Ascendente/metabolismo , Citocinas/metabolismo , Feminino , Humanos , Imunoglobulinas/análise , Lactente , Lactoferrina/administração & dosagem , Lactoferrina/efeitos adversos , Lactoferrina/sangue , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/metabolismo , Linfonodos/citologia , Linfonodos/crescimento & desenvolvimento , Linfonodos/imunologia , Linfonodos/metabolismo , Masculino , Baço/citologia , Baço/crescimento & desenvolvimento , Baço/imunologia , Baço/metabolismo , Sus scrofa , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/metabolismo , Regulação para CimaRESUMO
BACKGROUND: Kynurenic acid (KYNA), a tryptophan metabolite, was found in human saliva, gastric juice, bile, pancreatic juice and mucus of rat small intestine. METHODS: KYNA content in mucus aspirated from human caecum or colon ascendens and KYNA production in colon epithelial and cancer cells were determined using HPLC. Moreover, biological properties of KYNA and kynurenine aminotransferases (KATs) expression in colon epithelial and colon cancer cells were studied. RESULTS: Considerably higher KYNA concentration was detected in samples from patients diagnosed with colon carcinoma (269.40 ± 107.00 pmol/ml, N = 4), Adenoma tubulovillosum (200.50 ± 36.72, N = 10) or Adenoma tubulare (243.50 ± 38.09, N = 9) than in control group (82.22 ± 7.61 pmol/ml, N = 30). Moreover, colon epithelium CCD 841 CoTr cells actively synthesized KYNA in a concentration- and time-dependent manner. This process was decreased by aminooxyacetic acid and L-glutamate in opposite to 4-aminopyridine treatment. Interestingly, KYNA production in colon cancer cells (HT-29 1.39 ± 0.27, LS-180 1.18 ± 0.15 and Caco-2 4.21 ± 0.30 pmol/1 x 10(5) cells/2 h) was considerably higher in comparison to normal colon epithelial cells (0.70 ± 0.07 pmol/1 x 10(5) cells/2 h). However, KATs I and II were expressed at similar level in both colon epithelium and cancer cells. Furthermore, KYNA exerted an antiproliferative effect at higher micro- and millimolar concentrations against colon cancer cells with the IC(50) of 0.9, 0.2 and 1.2 mM for HT-29, LS-180 and Caco-2 cells, respectively. CONCLUSION: Summarizing, this is the first report presenting KYNA synthesis and KAT expression in colon derived normal and cancer cells.
Assuntos
Adenocarcinoma Mucinoso/metabolismo , Adenoma/metabolismo , Neoplasias do Colo/metabolismo , Ácido Cinurênico/metabolismo , Muco/metabolismo , Transaminases/metabolismo , Células CACO-2 , Ceco/enzimologia , Ceco/metabolismo , Proliferação de Células , Colo Ascendente/enzimologia , Colo Ascendente/metabolismo , Pólipos do Colo/metabolismo , Feminino , Células HT29 , Humanos , Ácido Cinurênico/análise , Masculino , Pessoa de Meia-Idade , Muco/químicaRESUMO
OBJECTIVE: This study aimed at investigating whether a differential estrogen receptor beta (ER-beta) expression between the colonic subsites could correspond to a modification in proliferation, apoptosis, and adhesion of the normal colonocytes. METHODS: ER-beta, Ki-67, Bcl-2, and E-cadherin expressions were investigated immunohistochemically, in normal epithelium biopsies from the ascending and the descending colon of 53 individuals, who underwent colonoscopy for the investigation of anemia and in whom no local pathology was identified. RESULTS: ER-beta immunoreactivity has been shown to be stronger at the superficial epithelium than the crypts' base, the difference being important only for the ascending colon. In addition, ER-beta expression was higher in the superficial epithelium of the ascending colon than that of the descending colon. The variations of ER-beta expression did not correspond to the alterations in Ki-67, Bcl-2, and E-cadherin expression. CONCLUSION: A subsite-specific variation of ER-beta expression has been shown in the normal colonic epithelium. This modulation of ER-beta might account for some well established specificities of colorectal cancer epidemiology like the right-sided predominance of the neoplasm in women and its gradual shift to more proximal sites over time.
Assuntos
Colo Ascendente/metabolismo , Colo Descendente/metabolismo , Neoplasias Colorretais/metabolismo , Receptor beta de Estrogênio/metabolismo , Mucosa Intestinal/metabolismo , Adulto , Idoso , Apoptose/fisiologia , Biópsia , Adesão Celular/fisiologia , Divisão Celular/fisiologia , Colo Ascendente/patologia , Colo Descendente/patologia , Colonoscopia , Neoplasias Colorretais/patologia , Feminino , Humanos , Imuno-Histoquímica , Mucosa Intestinal/patologia , Masculino , Pessoa de Meia-Idade , Valores de ReferênciaRESUMO
Plasma and tissue profiles of gastrointestinal hormones ghrelin and peptide YY (PYY) were investigated in different female rat reproductive states. Neither plasma nor tissue ghrelin concentrations were suppressed during pregnancy despite elevated leptin. The highest concentrations of stomach ghrelin were measured in late pregnancy. PYY concentrations in plasma, descending colon and rectum tissues were increased (P<0.001) throughout pregnancy and lactation. PYY peaked at day 5 of lactation in plasma, as well as descending colon and rectum tissues (proestrus vs day 5 of lactation: 25+/-3.0 pmol/l vs 55+/-8.0 pmol/l; 85+/-4.5 pmol/g wwt vs 418+/-45.0 pmol/g wwt; 23+/-3.0 pmol/g wwt vs 78+/-9.1 pmol/g wwt). This PYY peak was temporally associated with the luteinizing hormone peak on day 1 of lactation. Following weaning, dam adiposity and plasma leptin increased whereas ghrelin stomach peptide decreased. Relative PYY concentrations in the tissues of the gut varied in the different states suggesting regional alterations taking place in the colon. The ascending colon produced the highest concentrations in non-pregnant rats, the descending colon the highest concentrations during lactation with the pregnant rats and the dams postweaning in a transition state between. It is unclear what role the increased PYY in various tissues observed has during pregnancy and lactation as it would be expected to be reduced in these states of greatly increased appetite. PYY may have an influence on maternal dietary adaptation, intestinal hypertrophy and weight gain during pregnancy and lactation although it is still unclear precisely how it acts.
Assuntos
Mucosa Gástrica/metabolismo , Grelina/sangue , Grelina/metabolismo , Lactação/sangue , Lactação/metabolismo , Peptídeo YY/sangue , Peptídeo YY/metabolismo , Animais , Colo Ascendente/metabolismo , Colo Descendente/metabolismo , Feminino , Íleo/metabolismo , Leptina/sangue , Hormônio Luteinizante/sangue , Gravidez , Progesterona/sangue , Prolactina/sangue , Ratos , Reto/metabolismoRESUMO
OBJECTIVE: To study the effects of gut-derived endotoxin translocation and NF-kappaB activation on the aggravating mechanism of severe acute pancreatitis (SAP) and of treatment with pyrrolidine dithiocarbamate (PDTC) on rats with SAP. METHODS: SD rats were randomly divided into sham operation group (SO), SAP group, SAP + lipopolysaccharide(LPS) group, pyrrolidine dithiocarbamate (PDTC) treatment group and LPS group. Biochemical parameters and cytokines were examined in the serum. Multiple organs pathological slices were examined. Expression of NF-kappaB mRNA in the liver tissue was detected by RT-PCR. Activation of NF-kappaB by the method of streptomycin avidin-peroxidase (SP) and expression of NF-kappaB p65 protein and its binding activity were analyzed by Western blot and electrophoretic mobidity shift assay (EMSA). RESULTS: Compared with sham operation group, the concentration of TNF-alpha, alanine aminotransferase (ALT), and diamine oxidase (DAO) in serum significantly increased in SAP + LPS group (P < 0.05). Pathological changes were markedly observed in tissues and the expression of NF-kappaB mRNA in the liver significantly increased (P < 0.05) also, the activation of NF-kappaB and binding activity of NF-kappaB p65 protein in the liver markedly increased (P < 0.01) in SAP + LPS group. Treatment with PDTC markedly reduced concentration of ALT, DAO and TNF-alpha, and the expression of NF-kappaB, and the pathologic scores, as well as significantly decreased the expression of NF-kappaB p65 protein. CONCLUSION: The activation and overexpression of NF-kappaB may participate in the aggravating mechanism of SAP. Treatment with PDTC has a protective effect on multiple organs damage in SAP.