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AIMS: Cardiovascular events are the leading cause of death among patients with non-alcoholic fatty liver disease (NAFLD), but their relationship remains unclear. This study examined the association between coronary atherosclerosis and liver fibrosis, represented by the coronary artery calcification (CAC) score and non-invasive fibrosis markers, respectively. METHODS: Among 698 patients with chest pain or electrocardiographic abnormalities who underwent coronary computed tomography (CT) between April 2006 and March 2010, those with known liver disorders or history of emergency coronary angioplasty were excluded, leaving 366 patients for this study. Diagnosis of NAFLD was based on abdominal CT and history of alcohol consumption. Subjects with CAC of 100 AU or more were categorized into the high-risk group for cardiovascular events. Patient records were examined for clinical parameters including CAC score and non-invasive fibrosis marker FIB-4 index. RESULTS: Ninety-four patients (25.7%) had NAFLD. In this group, univariate analysis identified old age, high diastolic blood pressure, high liver to spleen ratio and high FIB-4 index as risk factors for cardiovascular events and multivariate analysis identified age of 66 years or older and FIB-4 index of 2.09 or more as the significant risk factors. For the observation period until August 2014, the cumulative proportion of PCI performance was significantly higher in patients with FIB-4 of 2.09 or more than those with FIB-4 of less than 2.09. CONCLUSION: The progression of arteriosclerosis and that of liver fibrosis may be associated in NAFLD patients. The FIB-4 index can be easily determined and thus can be a useful marker for predicting cardiovascular events in NAFLD patients.
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BACKGROUND AND AIMS: Esophageal visceral hypersensitivity has been proposed to be a pathogenesis of heartburn in nonerosive reflux disease (NERD), but its further mechanisms are unclear. Recently, it has been suggested that nociceptors and neuropeptides control sensory and pain mechanisms. Therefore, the objective of the present study was to estimate expression of acid-sensitive nociceptors such as transient receptor potential vanilloid 1 (TRPV1) and acid-sensing ion channel 3, protease-activated receptor 2 (PAR2), neuropeptides such as substance P and calcitonin-gene-related peptide, and their receptors such as neurokinin 1 receptor (NK1R) and receptor activity-modifying protein 1 in the esophageal mucosa of NERD patients. METHODS: Biopsy samples were taken from NERD patients and healthy control subjects without heartburn. The expression level of nociceptors, neuropeptides, and their receptors were assessed by real-time RT-PCR and enzyme immunoassay. Localization of substance P and CGRP in the esophageal mucosa was determined by immunohistochemical staining. RESULTS: Expression of mRNA for TRPV1 and PAR2 was significantly elevated in the esophageal mucosa of NERD patients. Substance P protein level and its receptor NK1R mRNA also increased in NERD patients. A positive correlation between the substance P protein level and reflux symptoms was observed. Immunohistochemical study revealed the presence of substance P-positive nerves in the lamina propria of the esophagus. CONCLUSIONS: These findings suggest that visceral hypersensitivity in NERD patients is involved in neurogenic inflammation showing the increase in both substance P release and NK1R expression, which may be associated with the activation of TRPV1 and PAR2.
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Refluxo Gastroesofágico/patologia , Neuropeptídeos/metabolismo , Nociceptores/metabolismo , Dor/metabolismo , Idoso , Esôfago/metabolismo , Feminino , Regulação da Expressão Gênica/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa/metabolismo , Neuropeptídeos/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Helicobacter pylori infection, which is a common cause of atrophic gastritis, has been reported to represent a causal factor increasing the vascular damage and consequent albuminuria. On the other hand, decreased serum pepsinogen (PG) I/II ratio can be used to assess gastric mucosal atrophy. To the best of our knowledge, there are no studies investigating the correlation between PG I/II ratio and diabetic nephropathy. Therefore, we investigated a relationship between PG I/II ratio and degree of urinary albumin excretion (UAE) in patients with type 2 diabetes. We evaluated relationships between PG I/II ratio and degree of UAE or estimated glomerular filtration rate as well as various factors, including age, body mass index, blood pressure, hemoglobin A1c, serum lipid concentrations, uric acid or C-reactive protein in 333 consecutive patients with type 2 diabetes. PG I/II ratio correlated positively with logarithm of UAE in all patients (r = 0.174, P = 0.0016) and in patients without Helicobacter pylori infection (r = 0.352, P < 0.0001). Multiple regression analysis identified that PG I/II ratio correlated independently with logarithm of UAE in all patients (ß = 0.264, P = 0.0005) and in patients without Helicobacter pylori infection (ß = 0.295, P = 0.0022). These data suggest that serum PG I/II ratio is correlated with diabetic nephropathy.
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Albuminúria/complicações , Diabetes Mellitus Tipo 2/complicações , Nefropatias Diabéticas/fisiopatologia , Rim/fisiopatologia , Pepsinogênio A/sangue , Pepsinogênio C/sangue , Idoso , Algoritmos , Anticorpos Antibacterianos/análise , Biomarcadores/sangue , Biomarcadores/urina , Estudos Transversais , Diabetes Mellitus Tipo 2/imunologia , Diabetes Mellitus Tipo 2/microbiologia , Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/epidemiologia , Nefropatias Diabéticas/urina , Feminino , Infecções por Helicobacter/complicações , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/imunologia , Humanos , Japão/epidemiologia , Rim/imunologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Índice de Gravidade de DoençaRESUMO
Atrophic gastritis is characterized by chronic inflammation of gastric mucosa by Helicobacter pylori infection and other factors. Helicobacter pylori infection has been linked to coronary artery disease. To our knowledge, however, no reports are available on the relationship between atrophic gastritis and coronary artery disease. In this study, we investigated the relationship between atrophic gastritis, which is diagnosed based on serum pepsinogen levels (pepsinogen I ≤ 70 ng/mL and pepsinogen I/II ratio ≤ 3.0), and the prevalence of coronary artery disease in general Japanese population. Among 2,633 study subjects, 531 subjects (20.2%) were diagnosed as atrophic gastritis. The prevalence of coronary artery disease was higher in the atrophic gastritis-positive group than that in the atrophic gastritis-negative group (5.8% vs 2.8%, p = 0.0005). Multiple logistic regression analysis demonstrated that atrophic gastritis was independently associated with coronary artery disease (odds ratio, 1.67; 95% confidence interval, 1.03-2.72), after adjustment for age, sex, obesity, hypertension, diabetes mellitus, dyslipidemia, hyperuricemia, and habits of smoking and drinking. These results suggest that atrophic gastritis is an independent risk factor for coronary artery disease. Chronic inflammation of gastric mucosa may be associated with the prevalence of coronary artery disease.
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OBJECTIVE: Under- and overnutrition are associated with a worse prognosis and constitute independent risk factors for morbidity and mortality. It is increasingly important to understand the factors that affect nutritional and metabolic statuses. The purpose of this study was to assess the relation between the pepsinogen I/II ratio and several biochemical markers. METHODS: A cross-sectional study was performed in 1985 subjects who underwent a health screening test. Subjects had no medications for hyperuricemia, dyslipidemia, diabetes mellitus, or hypertension. All subjects were classified into two groups. Subjects with a pepsinogen I/II ratio below 3 were defined as having atrophic gastritis. The relations between the pepsinogen I/II ratio and several biochemical markers, including total cholesterol, triacylglycerol, uric acid, cholinesterase, and glucose levels, were evaluated. RESULTS: The presence of atrophic gastritis was significantly associated with age, smoking status, alcohol consumption, body mass index, and triacylglycerol, uric acid, cholinesterase, and hemoglobin levels. Multiple linear regression analysis demonstrated that the pepsinogen I/II ratio was an independent determinant of glucose level (ß = 0.104, P < 0.0001), triacylglycerol level (ß = 0.072, P = 0.0014), uric acid level (ß = 0.048, P = 0.0138), and hemoglobin (ß = 0.037, P = 0.0429) after adjustments for age, sex, smoking status, alcohol consumption, and body mass index. CONCLUSION: The pepsinogen I/II ratio was related to glucose, triacylglycerol, and uric acid levels. Such an association fosters the idea that a decreased pepsinogen I/II ratio seems favorable for the prevention of overnutrition.
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Glicemia/metabolismo , Gastrite Atrófica/sangue , Obesidade/sangue , Pepsinogênio A/sangue , Pepsinogênio C/sangue , Triglicerídeos/sangue , Ácido Úrico/sangue , Adulto , Fatores Etários , Idoso , Consumo de Bebidas Alcoólicas , Biomarcadores/sangue , Índice de Massa Corporal , Colinesterases/sangue , Estudos Transversais , Feminino , Gastrite Atrófica/complicações , Nível de Saúde , Hemoglobinas/metabolismo , Humanos , Modelos Lineares , Masculino , Desnutrição , Programas de Rastreamento , Pessoa de Meia-Idade , Análise Multivariada , Obesidade/complicações , Obesidade/prevenção & controle , Prognóstico , Fatores de Risco , FumarRESUMO
We have investigated the flow birefringence property and assembly process of desmin, a muscle specific intermediate protein. Solution of non-polar desmin filaments showed birefringence when aligned in the sheared flow. The amount of birefringence of desmin filaments was considerably lower when compared with that of F-actin solution. Assembly of desmin from soluble state was followed by the birefringence measurements. At any desmin concentrations examined, the degree of flow birefringence increased rapidly just after the addition of the assembly buffer and reached a saturated level within 30 min. The time to reach half-maximal values of flow birefringence slightly but definitely depended on the initial soluble desmin concentrations. The plotting of the initial velocity of the assembly against the soluble desmin concentrations showed a slope of 1.4. This result suggested that the assembly process detected by flow birefringence measurements followed second-order kinetics, and the process corresponded to the second step of the three stage model for type III intermediate filament assembly proposed by Herrmann and his colleagues; the annealing of unit length filaments into filaments.
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Larval-to-adult myogenic conversion occurs in the dorsal muscle but not in the tail muscle during Xenopus laevis metamorphosis. To know the mechanism for tail-specific suppression of adult myogenesis, response character was compared between adult myogenic cells (Ad-cells) and larval tail myogenic cells (La-cells) to a Sonic hedgehog (Shh) inhibitor, notochord (Nc) cells, and spinal cord (SC) cells in vitro. Cyclopamine, an Shh inhibitor, suppressed the differentiation of cultured Ad (but not La) cells, suggesting the significance of Shh signaling in promoting adult myogenesis. To test the possibility that Shh-producing axial elements (notochord and spinal cord) regulate adult myogenesis, Ad-cells or La-cells were co-cultured with Nc or SC cells. The results showed that differentiation of Ad-cells were strongly inhibited by Nc cells but promoted by SC cells. If Ad-cells were "separately" co-cultured with Nc cells without direct cell-cell interactions, adult differentiation was not inhibited but rather promoted, suggesting that Nc cells have two roles, one is a short-range suppression and another is a long-range promotion for adult myogenesis. Immunohistochemical analysis showed both notochord and spinal cord express the N-terminal Shh fragment throughout metamorphosis. The "spinal cord-promotion" and long-range effect by Nc cells on adult myogenesis is thus involved in Shh signaling, while the signaling concerning the short-range "Nc suppression" will be determined by future studies. Interestingly, these effects, "Nc suppression" and "SC promotion" were not observed for La-cells. Situation where the spinal cord/notochord cross-sectional ratio is quite larger in tadpole trunk than in the tail seems to contribute to trunk-specific promotion and tail-specific suppression of adult myogenesis during Xenopus metamorphosis.
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Diferenciação Celular/fisiologia , Metamorfose Biológica/fisiologia , Desenvolvimento Muscular/fisiologia , Notocorda/citologia , Medula Espinal/citologia , Xenopus laevis/anatomia & histologia , Xenopus laevis/fisiologia , Animais , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Regulação da Expressão Gênica no Desenvolvimento , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Larva/anatomia & histologia , Larva/fisiologia , Desenvolvimento Muscular/efeitos dos fármacos , Transdução de Sinais/fisiologia , Cauda/citologia , Cauda/fisiologia , Alcaloides de Veratrum/farmacologia , Proteínas de Xenopus/genética , Proteínas de Xenopus/metabolismoRESUMO
N-terminal actin-binding domain of α-actinin is connected to central rod domain through flexible neck region that is susceptible to proteolysis. It is suggested that the neck region assumes variable orientations by actin binding. In order to examine the effect of actin binding to α-actinin, we carried out limited digestion of α-actinin by chymotrypsin in the presence and absence of F-actin. Although the cleavage process was retarded when bound to F-actin, digestion to 32 kDa-head and 55 kDa-rod domains occurred through the same intermediate products as the digestion in the absence of F-actin. N-terminal sequencing of 55 kDa-fragment showed the neck region was cleaved at 276-Leu. The cleavage site was not affected by binding to F-actin nor ionic strength of the solvent. It was also indicated that α-actinin was cleaved at 15-Tyr by chymotrypsin. Quantitation of the cleavage products by densitometry of the SDS-gels suggested the conformational change of α-actinin at domain-connecting regions by F-actin binding.
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Isoforms of myosin heavy chain and tropomyosin convert during metamorphosis of Xenopus laevis with larval-to-adult remodeling of dorsal muscle (Nishikawa and Hayashi, 1994 , Dev. Biol. 165: 86-94). In the present study, other markers for muscle remodeling during metamorphosis were determined by SDS-PAGE analysis. The amounts of twelve muscle proteins changed remarkably during metamorphosis. Among these, a 54-kDa molecule was found to be desmin, and the relative content/total proteins decreased dramatically through metamorphosis. In hindlimb muscle, desmin content increased fourfold during prometamorphosis, when myoblast proliferation and fusion occurred. With further myotube maturation, this content decreased by 1/2 while that of muscle actin continued to increase. Thus, desmin up- and down-regulation in hindlimbs mark early and late phases of myogenesis, respectively. In tall muscle, the desmin content decreased continuously to (1/8) before and during metamorphosis, due to tall muscle growth and maturation. In dorsal muscle, three desmin changes occurred: a pre-metamorphic decrease, a transient increase at prometamorphosis, and a rapid decrease at the climax stage. Immunohistochemical analysis showed desmin+ cells to be present between young (adult-type) myotubes and replicating (PCNA+) cells in dorsal muscles, correlating the transient desmin upregulation in dorsal muscle with the initiation of adult-type myogenesis. After the upregulation, dorsal muscle desmin decreased to (1/8). This rapid down-regulation was replicated by administration of triiodothyronine (T3) to tadpoles, suggesting a significant role for T3 in dorsal muscle remodeling during metamorphosis. Collectively, these results show that analysis of desmin expression and PCNA-immunohistochemistry are good tools for determining the sites and timing of larval-to-adult muscle remodeling during Xenopus laevis metamorphosis.
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Desmina/metabolismo , Metamorfose Biológica/fisiologia , Desenvolvimento Muscular/fisiologia , Músculo Esquelético/crescimento & desenvolvimento , Xenopus laevis/crescimento & desenvolvimento , Animais , Anticorpos , Desmina/genética , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Regulação da Expressão Gênica no Desenvolvimento/fisiologia , Larva/crescimento & desenvolvimento , Larva/metabolismo , Tiroxina/farmacologiaRESUMO
BACKGROUND AND AIM: Induction of inducible nitric oxide synthase (iNOS) may be involved in carcinogenesis of the stomach, because nitric oxide (NO) derived from iNOS can exert DNA damage and post-transcriptional modification of target proteins. In the present study, we investigated the correlation between endoscopic findings and iNOS mRNA expression/NO-modified proteins in the gastric mucosa. METHODS: Fifty patients were prospectively selected from subjects who underwent upper gastrointestinal chromoendoscopy screening for abdominal complaints. The Helicobacter pylori (H. pylori) status of patients was determined by anti-H. pylori IgG antibody levels. We classified the mucosal area of the fundus as F0, fine small granules; F1, edematous large granules without a sulcus between granules; F2, reduced-size granules with a sulcus between granules; and F3, irregular-sized granules with extended sulcus between granules. Gastritis was graded using the visual analog scale of the Updated Sydney System. The expression of interleukin (IL)-8 and iNOS mRNA was assayed in gastric biopsy specimens by reverse transcription-polymerase chain reaction. NO-modified proteins were analyzed by Western blotting using novel monoclonal antibodies against nitrotyrosine. RESULTS: A total of 91.7% (11/12) of the F0 group was H. pylori-negative, whereas 94.7% (36/38) of the F1-3 groups was H. pylori-positive. Spearman's analysis showed good correlation between the endoscopic grading and the score of chronic inflammation (r=0.764) and glandular atrophy (r=0.751). The expression of IL-8 mRNA was significantly increased in F1, F2, and F3 cases compared with the F0 group, with no significant differences among them. iNOS mRNA was significantly increased in the F3 group compared with the other groups, with increased nitration of tyrosine residues of proteins. CONCLUSION: The proposed classification by chromoendoscopy is useful for screening patients for atrophic and iNOS-expressing gastric mucosa with NO-modified proteins in H. pylori-associated atrophic gastric mucosa.
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Mucosa Gástrica/enzimologia , Gastrite Atrófica/enzimologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/patogenicidade , Óxido Nítrico Sintase Tipo II/análise , Tirosina/análogos & derivados , Anticorpos Antibacterianos/sangue , Atrofia , Biomarcadores/análise , Mucosa Gástrica/microbiologia , Mucosa Gástrica/patologia , Gastrite Atrófica/microbiologia , Gastrite Atrófica/patologia , Gastroscopia/métodos , Infecções por Helicobacter/enzimologia , Infecções por Helicobacter/patologia , Helicobacter pylori/imunologia , Humanos , Interleucina-8/análise , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/genética , Estudos Prospectivos , Proteínas/metabolismo , RNA Mensageiro/análise , Índice de Gravidade de Doença , Tirosina/análiseRESUMO
We have reported that rebamipide, a gastroprotective drug, suppresses indomethacin-induced gastric mucosal injury in humans and rats. However, the mechanisms of the cytoprotective actions of rebamipide have not been fully addressed. In the present study, we determined mRNA expression profile of the gastric mucosa treated with indomethacin in rats, and investigated the cytoprotective effects of rebamipide against indomethacin-induced injury with a high-density oligonucleotide array (Rat Toxicology U34 GeneChip array). Gastric epithelial cells were obtained by laser-assisted microdissection. Data analysis was performed with a GeneChip Operating Software, GeneSpring software 7.0, and Ingenuity Pathway Analysis. Among 1,031 probes, the expression of 160 probes (15.5%) showed at least 2.0-fold up-regulation (158 probes) and down-regulation (2 probes) 2 h after indomethacin administration in comparison with the vehicle-treated rats. The pathway analysis of the up-regulated 123 probes identified the network with a highly significant score, which consisted of known clusters of cell death, cancer, and endocrine system disorders. We succeeded in listing 10 genes that were up-regulated by the treatment with indomethacin and that were down-regulated by rebamipide, including growth arrest and DNA damage-induced 45alpha. In conclusion, we demonstrated that cell death, especially apoptosis, pathway is involved in the pathogenesis of indomethacin-induced gastric mucosal injury, and that inhibition of apoptosis-related genes is possibly important for the cytoprotective effect of rebamipide against this injury.
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Recent studies have shown that esophageal mucosal inflammatory response is involved in the pathophysiology of gastro-esophageal reflux disease. The aim of the present study was to identify specific gene expression profiles of the esophageal mucosa in a rat model of combined-type chronic reflux esophagitis. Esophagogastroduodenal anastomosis was carried out in male Wistar rats by anastomosing the jejunum to the gastroesophageal junction under diethyl-ether inhalation anesthesia. Esophageal epithelial cells were obtained from esophagi of rats by laser capture microdissection. Preparation of cRNA and target hybridization were performed according to the Affymetrix GeneChip eukaryotic small sample target labeling assay protocol. The gene expression profile was evaluated by the rat toxicology U34 GeneChip. Array data analysis was carried out using Affymetrix GeneChip operating software, ingenuity pathway analysis software, and Gene Springs software. A comparison between esophagitis and sham-operated rats 2 weeks after the operation revealed that 368 probes (36%) were significantly affected, i.e. 185 probes were up-regulated, and 183 probes were down-regulated, both at levels of at least 1.5-fold in the esophagitis rats. Ingenuity signal analysis of 207 affected probes revealed the interleukin-6 signaling pathway as the most significantly affected caronical pathway. In addition, the expression of many genes associated with cytokine and transcription factor was enhanced in the esophagitis rats. This transcriptome approach provided insight into genes and putative genetic pathways thought to be affected by stimulation with gastroduodenal refluxates.
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Esofagite/genética , Refluxo Gastroesofágico/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Animais , Epitélio/metabolismo , Epitélio/patologia , Esôfago/metabolismo , Esôfago/patologia , Masculino , Mucosa/metabolismo , Mucosa/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Ratos , Ratos Wistar , Transcrição GênicaRESUMO
We have demonstrated that astaxanthin reduces glomerular oxidative stress as well as inhibits the increase in urinary albumin in diabetic db/db mice. The aim of the present study was to determine the gene expression patterns in the glomerular cells of the diabetic mouse kidney, and to investigate the effects of astaxanthin on the expression of these genes using a high-density DNA microarray. The diet administered to the astaxanthin-supplementation group was prepared by mixing a control powder with astaxanthin at a concentration of 0.02%. Glomerular cells were obtained from the kidneys of mice by laser capture microdissection. Preparation of cRNA and target hybridization were performed according to the Affymetrix GeneChip eukaryotic small sample target labeling assay protocol. The gene expression profile was evaluated by the mouse expression set 430A GeneChip. Array data analysis was carried out using Affymetrix GeneChip operating and Ingenuity Pathway analysis software. Comparison between diabetic db/db and non-diabetic db/m mice revealed that 779 probes (3.1%) were significantly affected, i.e. 550 probes were up-regulated, and 229 probes were down-regulated, both at levels of >/=1.5-fold in the diabetic mice. Ingenuity signal analysis of 550 up-regulated probes revealed the mitochondrial oxidative phosphorylation pathway as the most significantly affected caronical pathway. The affected genes were associated with complexes I, III, and IV located on the mitochondrial inner membrane, and the expression levels of these genes were decreased in mice treated with astaxanthin as compared to the levels in the control mice. In addition, the expression of many genes associated with oxidative stress, collagen synthesis, and transforming growth factor-beta signaling was enhanced in the diabetic mice, and this enhancement was slightly inhibited in the astaxanthin-treated mice. In conclusion, this genome-wide nutrigenomics approach provided insight into genes and putative genetic pathways that are thought to be affected by stimulation by high-glucose concentrations. In addition, the present approach may help us gain a better understanding of the genes and pathways involved in the anti-diabetic mechanism of astaxanthin.
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Diabetes Mellitus Tipo 2/tratamento farmacológico , Perfilação da Expressão Gênica , Glomérulos Renais/efeitos dos fármacos , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Animais , Diabetes Mellitus Tipo 2/genética , Feminino , Expressão Gênica/efeitos dos fármacos , Expressão Gênica/genética , Glomérulos Renais/metabolismo , Células Mesangiais/efeitos dos fármacos , Células Mesangiais/metabolismo , Camundongos , Camundongos Obesos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Xantofilas/administração & dosagem , Xantofilas/farmacologia , Xantofilas/uso terapêuticoRESUMO
OBJECTIVE: Mast cell tryptase has been proposed to be involved in the pathogenesis of human inflammatory bowel disease (IBD). Recently, it was reported that a low dose of nafamostat mesilate (NM), a serine protease inhibitor that is widely used to treat disseminated intravascular coagulation (DIC) and acute pancreatitis, can selectively inhibit human tryptase activity. The aim of this study was to investigate the anti-inflammatory effects of NM on experimental colitis in rats. MATERIAL AND METHODS: Colitis was induced in male Wistar rats using an enema of trinitrobenzene sulfonic acid (TNBS) dissolved in 50% ethanol. NM or 5-aminosalicylic acid (5-ASA), foundation therapy for mild-to-moderate IBD, was administered via the anus once a day on each of the 6 days after administration of TNBS. Colonic inflammation was assessed 1 week after TNBS administration. RESULTS: Intracolonic administration of TNBS resulted in the infiltration of numerous tryptase-positive cells in the colonic mucosa. The colonic mucosal injury induced by TNBS was significantly decreased by treatment with NM or 5-ASA. The increases in thiobarbituric acid-reactive substances (TBA-RS), myeloperoxidase (MPO) activity, tumor necrosis factor (TNF)-alpha and cytokine-induced neutrophil chemoattractants-1 (CINC-1) in the colonic mucosa were inhibited in the NM group and the 5-ASA group, without significant differences between them. CONCLUSIONS: These results indicate that a low dose of NM can inhibit the colonic mucosal inflammation induced by TNBS in rats, which suggests that anti-tryptase therapy using low doses of NM has excellent potential to become a new therapeutic strategy for IBD.
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Anti-Inflamatórios não Esteroides/uso terapêutico , Colite/tratamento farmacológico , Guanidinas/uso terapêutico , Inibidores de Serina Proteinase/uso terapêutico , Triptases/antagonistas & inibidores , Ácidos Aminossalicílicos/uso terapêutico , Animais , Benzamidinas , Quimiocina CXCL1 , Quimiocinas CXC/análise , Colite/metabolismo , Colite/patologia , Colo/metabolismo , Colo/patologia , Mucosa Intestinal/patologia , Masculino , Ratos , Ratos Wistar , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Ácido Trinitrobenzenossulfônico , Triptases/análise , Fator de Necrose Tumoral alfa/análiseRESUMO
BACKGROUND: Reflux of the duodenal contents with gastric acid has been reported to contribute to the development of esophageal mucosal damage and inflammation. Recent studies show that pancreatic trypsin can stimulate the production of inflammatory mediators, including chemokines and prostaglandins from human esophageal epithelial cells in vitro. The aim of the present study was to investigate the role of pancreatic trypsin in the pathogenesis of chronic esophageal inflammation induced by gastroduodenal reflux in rats. METHODS: Esophagogastroduodenal anastomosis was carried out in male Wistar rats by anastomosing the jejunum to the gastroesophageal junction under diethyl ether inhalation anesthesia. The animals undergoing surgery were treated with the control diet, rabeprazole sodium, nizatidine, ecabet sodium, camostat mesilate (CMM), ONO-1714, a specific inducible nitric oxide synthase (iNOS) inhibitor, or meloxicam, a selective cyclooxygenase-2 (COX-2) inhibitor. Esophageal injury was evaluated by macroscopic and microscopic findings, and mRNA expression for CINC-1, COX-2, and iNOS was determined by real-time polymerase chain reaction (PCR). Trypsin activity within the esophageal lumen was measured 2 weeks after surgery, and the expression of protease-activated receptor (PAR)-1 and -2 was confirmed by reverse transcription (RT)-PCR. RESULTS: At 8 weeks after surgery, gastroduodenal reflux induced esophageal erosions and ulcer formation as well as marked thickening of the esophageal wall. Histological study showed an increase of thickness of the esophageal mucosa, hyperplasia of the epidermis and basal cells, ulcer formation, and marked infiltration of inflammatory cells. The macroscopic ulcer score and histological ulcer length were significantly reduced by treatment with rabeprazole or CMM but not by nizatidine or ecabet sodium, compared with each control. Rabeprazole, nizatidine, or ecabet sodium did not affect the severity of mucosal hyperplastic scores or histological parameters in esophagitis. In contrast, the CMM group showed a significant decrease in the mucosal hyperplastic and inflammatory scores. The enhanced expression of CINC-1, COX-2, and iNOS mRNA in the control group was also markedly inhibited in the CMM-treated group. ONO-1714 or meloxicam treatment significantly reduced the macroscopic scores of ulcer and hyperplasia. The trypsin activity in the esophageal lumen was significantly increased in the control diet group, and this increase was significantly inhibited in the CMM-treated group. The expression of PAR-1 and -2 mRNA was confirmed in rat esophageal epithelium. CONCLUSIONS: With this model, we have demonstrated that CMM significantly reduces inflammation and hyperplasia in the esophageal mucosa. These results indicate that trypsin, which is primarily inhibited by CMM, plays an important role in the mucosal damage induced by gastroduodenal reflux and that it can be a therapeutic target in patients with gastroduodenal reflux esophagitis.
Assuntos
Refluxo Duodenogástrico/complicações , Refluxo Duodenogástrico/enzimologia , Esofagite/enzimologia , Esofagite/etiologia , Tripsina/metabolismo , Animais , Quimiocina CXCL1 , Quimiocinas CXC/biossíntese , Doença Crônica , Ciclo-Oxigenase 2/biossíntese , Ciclo-Oxigenase 2/efeitos dos fármacos , Inibidores de Ciclo-Oxigenase/farmacologia , Modelos Animais de Doenças , Refluxo Duodenogástrico/patologia , Esofagite/patologia , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Masculino , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/biossíntese , Óxido Nítrico Sintase Tipo II/efeitos dos fármacos , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Receptor PAR-1/biossíntese , Receptor PAR-1/efeitos dos fármacos , Receptor PAR-2/biossíntese , Receptor PAR-2/efeitos dos fármacos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Inibidores da Tripsina/farmacologiaRESUMO
AIM: To investigate the protective effect of rosuvastatin on ischemia-reperfusion (I-R)-induced small intestinal injury and inflammation in rats, and to determine the effect of this agent on the expression of endothelial nitric oxide synthase (eNOS) protein. METHODS: Intestinal damage was induced in male Sprague-Dawley rats by clamping both the superior mesenteric artery and the celiac trunk for 30 min, followed by reperfusion for 60 min. Rosuvastatin dissolved in physiological saline was administered intraperitoneally 60 min before ischemia. The severity of the intestinal mucosal injury and inflammation were evaluated by several biochemical markers, as well as by histological findings. The protein levels of eNOS were determined by Western blot. RESULTS: The levels of both intraluminal hemoglobin and protein, as indices of mucosal damage, were significantly increased in the I-R group compared with those in the sham-operated group. These increases, however, were significantly inhibited by treatment with rosuvastatin in a dose-dependent manner. The protective effects of rosuvastatin were also confirmed by histological findings. Exposure of the small intestine to I-R resulted in mucosal inflammation characterized by significant increases in thiobarbituric acid-reactive substances, tissue-associated myeloperoxidase activity, and the mucosal contents of rat cytokine-induced neutrophil chemoattractant-1 (CINC-1) and tumor necrosis factor-alpha (TNF-alpha). These increases in inflammatory parameters after I-R were significantly inhibited by pretreatment with rosuvastatin at a dose of 10 mg/kg. Furthermore, mRNA expression of CINC-1 and TNF-alpha was increased after I-R, and this increase was also inhibited by rosuvastatin. The mucosal protein levels of eNOS decreased during I-R, but were preserved in rats treated with rosuvastatin. CONCLUSION: Rosuvastatin inhibits rat intestinal injury and inflammation induced by I-R, and its protection is associated with the preservation of eNOS protein.
Assuntos
Fluorbenzenos/farmacologia , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Intestinos/irrigação sanguínea , Óxido Nítrico Sintase Tipo III/análise , Pirimidinas/farmacologia , Traumatismo por Reperfusão/prevenção & controle , Sulfonamidas/farmacologia , Animais , Quimiocina CXCL1 , Quimiocinas CXC/genética , Fluorbenzenos/uso terapêutico , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Pirimidinas/uso terapêutico , RNA Mensageiro/análise , Ratos , Ratos Sprague-Dawley , Traumatismo por Reperfusão/enzimologia , Rosuvastatina Cálcica , Sulfonamidas/uso terapêutico , Fator de Necrose Tumoral alfa/genéticaRESUMO
Lipid peroxidation mediated by oxygen free radicals plays an important role in the pathogenesis of inflammatory bowel disease (IBD). Vitamin E is a lipid-soluble antioxidant and is generally considered to protect against lipid peroxidation of the cell membrane and to scavenge singlet oxygen and superoxide anion radical. Therefore, vitamin E or its derivatives are expected to have particular application for patients suffering from IBD. The aim of this study was to investigate the antioxidative effects of the water-soluble vitamin E derivative, 2-(alpha-D-glucopyranosyl)methyl-2,5,7,8-tetra-methylchroman-6-ol(TMG), on the therapy of experimental colitis in rats. Colitis was induced in male Wistar rats weighing 200 g using an enema of trinitrobenzene sulfonic acid (TNBS) dissolved in 50% ethanol; TMG dissolved in physiological saline was injected intra-peritoneally every day from 24 h after the enema of TNBS. The damage score, wet weight of the colon, and increase in body weight were estimated 1 week after the enema of TNBS. Thiobarbituric acid-reactive substances (TBA-RS), an index of lipid peroxidation, and tissue-associated myeloperoxidase (MPO) activity in the colonic mucosa were measured 1 week after the induction of colitis. As a result, increase in body weight was inhibited by the induction of colitis, although the inhibition was reduced in the group treated with TMG. The damage score, wet weight, TBA-RS and MPO activity were increased significantly in the colitis group; however, they were inhibited by the administration of TMG. These results suggest that TMG is effective for the treatment of colitis in rats induced by TNBS. In the future, TMG could be a new therapeutic agent for IBD.
Assuntos
Cromanos/química , Cromanos/uso terapêutico , Colite/tratamento farmacológico , Glicosídeos/química , Glicosídeos/uso terapêutico , Vitamina E/análogos & derivados , Animais , Peso Corporal/efeitos dos fármacos , Cromanos/farmacologia , Colite/induzido quimicamente , Colo/patologia , Citocinas/metabolismo , Glicosídeos/farmacologia , Inflamação , Mucosa Intestinal/patologia , Masculino , Peroxidase/metabolismo , Ratos , Ratos Wistar , Solubilidade , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismo , Ácido TrinitrobenzenossulfônicoRESUMO
The spread of capsule endoscopy has led to a focus on small intestinal injury induced by non-steroidal anti-inflammatory drugs (NSAIDs). However, it has been proposed that proton pump inhibitors (PPI), a strong anti-secretary agent, have anti-inflammatory action beyond acid suppression. Therefore, we evaluated the biological effects of lansoprazole, a PPI used in the clinical area, in the setting of experimental rat non-steroidal anti-inflammatory drug-induced enteritis. The animals were given indomethacin subcutaneously and the intestinal mucosa was examined 24 h later. Lansoprazole was given subcutaneously just after following indomethacin injection. Single administration of indomethacin at 10 mg/kg provoked severe hemorrhagic lesions in the small intestine, mostly the jejunum and ileum. The levels of thiobarbituric acid-reactive substances (TBARS), the myeloperoxidase (MPO) activity and the content of cytokine-induced neutrophil chemoattractant-1 (CINC-1) in the intestinal mucosa significantly increased in indomethacin-treated groups compared with the sham-operated groups. The development of intestinal lesions in response to indomethacin was dose-dependently prevented by lansoprazole at a dose of 5 mg/kg together with significant suppression of the increased level of TBARS, MPO activities and CINC-1 in the small bowel. Furthermore, the increased CINC-1 mRNA expression after administration of indomethacin was also inhibited by treatment with lansoprazole. These results suggest that lansoprazole administered exogenously prevented the small intestine against indomethacin-induced damage, the action being dependent on its anti-inflammatory and anti-oxidative responses. This evidence supports the theory that PPI have an expanding role beyond acid suppression.
Assuntos
Anti-Inflamatórios não Esteroides/toxicidade , Enterite , Indometacina/toxicidade , Omeprazol/análogos & derivados , Inibidores da Bomba de Prótons , 2-Piridinilmetilsulfinilbenzimidazóis , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Quimiocina CXCL1 , Quimiocinas CXC/genética , Quimiocinas CXC/metabolismo , Enterite/induzido quimicamente , Enterite/tratamento farmacológico , Enterite/patologia , Indometacina/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/patologia , Intestino Delgado/anatomia & histologia , Intestino Delgado/metabolismo , Lansoprazol , Masculino , Omeprazol/metabolismo , Omeprazol/uso terapêutico , Peroxidase/metabolismo , Bombas de Próton/metabolismo , Ratos , Ratos Wistar , Substâncias Reativas com Ácido Tiobarbitúrico/metabolismoRESUMO
Partially hydrolyzed guar gum (PHGG), a water-soluble dietary fiber produced by a controlled partial enzymatic hydrolysis of guar gum beans, has various physiological actions. The aim of the present study was to elucidate the beneficial effects of PHGG on colonic mucosal damage and on the inflammatory response in a dextran sulfate sodium (DSS) colitis model. After 2 weeks of prefeeding of PHGG, acute colitis was induced with 8% DSS in female BALB/c mice. Colonic mucosal inflammation was evaluated clinically, biochemically and histologically. Mucosal protein contents and mRNA levels of tumor necrosis factor-alpha (TNF-alpha) were determined by immunoassay and reverse transcription polymerase chain reaction. Disease activity scores determined by weight loss, stool consistency and blood in stool in DSS-treated mice were significantly lower in the PHGG-treated mice compared with the control mice. Shortening of the colon was significantly reversed by PHGG. Histological study also showed a reduced infiltration of inflammatory cells, especially neutrophils, and mucosal cell disruption in PHGG-treated mice compared with the control mice. The increases in tissue-associated myeloperoxidase activity and thiobarbituric acid-reactive substances after DSS administration were both significantly inhibited by pretreatment with PHGG. Partially hydrolyzed guar gum also inhibited increases in intestinal TNF-alpha protein and mRNA expression after DSS administration, respectively. These results suggest that chronic ingestion of PHGG prevents the development of DSS-induced colitis in mice via the inhibition of mucosal inflammatory response.
Assuntos
Colite/tratamento farmacológico , Colo/patologia , Sulfato de Dextrana , Galactanos/administração & dosagem , Galactanos/metabolismo , Mananas/administração & dosagem , Mananas/metabolismo , Animais , Colite/induzido quimicamente , Colite/patologia , Contagem de Colônia Microbiana , Fezes/microbiologia , Feminino , Hidrólise , Mucosa Intestinal/química , Mucosa Intestinal/patologia , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/patologia , Peroxidase/metabolismo , Gomas Vegetais , RNA Mensageiro/análise , Substâncias Reativas com Ácido Tiobarbitúrico/análise , Fator de Necrose Tumoral alfa/análise , Fator de Necrose Tumoral alfa/genéticaRESUMO
Oxygen radical-mediated lipid peroxidation and neutrophil activation may be involved in the development of gastric mucosal injury induced by non-steroidal anti-inflammatory drugs (NSAIDs). Vitamin E is one of the lipid-soluble antioxidants and is generally considered to protect against lipid peroxidation of the cell membrane and to scavenge singlet oxygen and superoxide anion radicals. Our object was to investigate the antioxidative effects of water-soluble vitamin E derivative, 2-(alpha-D-glucopyranosyl)methyl-2,5,7,8-tetra-methylchroman-6-ol (TMG), on aspirin-induced gastric mucosal injury in rats. Gastric injury was induced by intragastric administration of aspirin and 0.15 N HCl in male Sprague-Dawley rats. TMG dissolved in physiological saline was injected intraperitoneally 0.5 h before the aspirin administration. The intragastric administration of acidified aspirin induced hyperemia and hemorragic erosions in rat stomach. The increase in total area of gastric erosions was reduced by pretreatment with TMG in a dose-dependent manner. The increases of thiobarbituric acid-reactive substances (TBA-RS) and myeloperoxidase (MPO) activity 3 h after aspirin administration were significantly inhibited by pretreatment with TMG. The gastric concentration of cytokine-induced neutrophil chemoattractants-1 (CINC-1) increased after aspirin administration, and the increase was also inhibited by pretreatment with TMG. These results suggest that TMG is effective for the treatment of aspirin-induced gastric injury. This anti-inflammatory effect of TMG seems to be related to impairment of lipid peroxidation, neutrophil function and cytokine production in gastric mucosa.