RESUMO
The use of various herbs and their compounds has been a strategy widely used in the fight against various human diseases. For example, rosmarinic acid, a bioactive phenolic compound commonly found in Rosemary plants (Rosmarinus officinalis Labiatae), has multiple therapeutic benefits in different diseases, such as cancer. Therefore, the study aimed to evaluate in silico and in vitro the inhibition potential of the enzyme Elastase from the porcine pancreas by rosmarinic acid isolated from the plant species R. officinalis Linn. Through Molecular Docking, the mechanism of action was investigated. In addition, rosmarinic acid presented a range of 5-60 µg/mL and significantly inhibited Elastase. At 60 µg/mL, there was an inhibition of 55% on the enzymatic activity. The results demonstrate the inhibition of Elastase by rosmarinic acid, which can lead to the development of new enzyme inhibitors that can be an inspiration for developing various drugs, including anticancer drugs.
Assuntos
Ácido Rosmarínico , Rosmarinus , Humanos , Elastase Pancreática , Simulação de Acoplamento Molecular , Extratos Vegetais/farmacologia , Cinamatos/farmacologia , Depsídeos/farmacologiaRESUMO
Chronic obstructive pulmonary disease (COPD) is an important health care issue, and IL-17 can modulate inflammatory responses. We evaluated preventive and therapeutic effect of anti-interleukin (IL)-17 in a model of lung injury induced by elastase, using 32 male C57Bl6 mice, divided into 4 groups: SAL, ELASTASE CONTROL (EC), ELASTASE + PREVENTIVE ANTI-IL-17 (EP), and ELASTASE + THERAPEUTIC ANTI-IL-17 (ET). On the 29th day, animals were anesthetized with thiopental, tracheotomized, and placed on a ventilator to evaluate lung mechanical, exhaled nitric oxide (eNO), and total cells of bronchoalveolar lavage fluid was collected. We performed histological techniques, and linear mean intercept (Lm) was analyzed. Both treatments with anti-IL-17 decreased respiratory resistance and elastance, airway resistance, elastance of pulmonary parenchyma, eNO, and Lm compared with EC. There was reduction in total cells and macrophages in ET compared with EC. Both treatments decreased nuclear factor-кB, inducible nitric oxide synthase, matrix metalloproteinase (MMP)-9, MMP-12, transforming growth factor-ß, tumor necrosis factor-α, neutrophils, IL-1ß, isoprostane, and IL-17 in airways and alveolar septa; collagen fibers, decorin and lumican in airways; and elastic fibers and fibronectin in alveolar septa compared with EC. There was reduction of collagen fibers in alveolar septa and biglycan in airways in EP and a reduction of eNO synthase in airways in ET. In conclusion, both treatments with anti-IL-17 contributed to improve most of parameters evaluated in inflammation and extracellular matrix remodeling in this model of lung injury.
Assuntos
Interleucina-17/metabolismo , Lesão Pulmonar/metabolismo , Pulmão/metabolismo , Elastase Pancreática/metabolismo , Animais , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Inflamação/metabolismo , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neutrófilos/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Doença Pulmonar Obstrutiva Crônica/metabolismoRESUMO
Introdução: fibrose cística (FC) é uma doença genética que culmina em alterações na proteína transmembrana CFTR, resultando na produção de muco mais espesso em diversos órgãos, destacando-se os tratos respiratório e digestório. A insuficiência pancreática (IP) acomete até 95% dos pacientes com FC. Objetivos: determinar a frequência de IP através da dosagem de elastase fecal-1 (EF-1) e compará-la com o genótipo de pacientes com FC assistidos em um centro de referência. Metodologia: foi realizado um estudo transversal, incluindo-se pacientes com FC de 0 a 20 anos. Após a inclusão dos sujeitos à pesquisa, foi realizada consulta ao prontuário para a obtenção de dados clinicos e demograficos e amostras de fezes foram obtidas para dosagem da (EF-1). Os pacientes foram submetidos à análise molecular das mutações por métodos convencionais, através da extração do DNA em sangue periférico. Quando duas mutações patológicas não foram identificadas, o sequenciamento de nova geração com utilização da plataforma Illumina HiSeq foi realizado em amostras da mucosa oral. Resultados: foram incluídos 31 pacientes, 17 (54,8%) do sexo feminino, mediana de idade de 10 anos, e apenas um paciente foi classificado como branco. Vinte e dois (70,9%) pacientes apresentaram dosagem de EF-1 inferior a 200 µg/g, compatível com o diagnóstico de IP. Destes, 21 (95,4%) apresentaram dosagem de EF-1 menor ou igual a 15µg/g, caracteristica de IP grave. Todos os pacientes com IP apresentavam duas mutações de classes I a III. Conclusão: a IP foi identificada em 70% dos pacientes, ocorrendo em todos os pacientes com duas mutações de classe I-III.
Introduction: cystic fibrosis (CF) is a genetic disease that culminates in alterations in the CFTR transmembrane protein, resulting in the production of thicker mucus in various organs, especially the respiratory and digestive tract. Pancreatic insufficiency (PI) affects up to 95% of CF patients. Objectives: To determine the frequency of PI by measuring fecal elastase-1 (FE-1) and comparing it with the genotype of CF patients assisted at a referral Center. Methodology: a cross-sectional study was conducted, including patients with CF from 0 to 20 years. After the inclusion of the subjects to the research, medical records were consulted to obtain clinical and demographic data and stool samples were obtained for the measurement of (FE-1). Patients were submitted to molecular analysis of mutations by conventional methods by DNA extraction in peripheral blood. When two pathological mutations were not identified, next-generation sequencing using the Illumina HiSeq platform was performed on oral mucosa samples. Results: thirty one patients were included, 17 (54.8%) female, median age 10 years, and only one patient was classified as white. Twenty-two (70.9%) patients had an FE-1 dosage of less than 200 µg / g, compatible with the diagnosis of pancreatic insufficiency (PI). Of these, 21 (95,4%) had an EF-1 dosage less than or equal to 15µg / g, characteristic of severe PI. All patients with two mutations class I to III were PI. Conclusion: PI was identified in 70% of patients, occurring in all patients with class I-III mutations.
Assuntos
Humanos , Masculino , Feminino , Criança , Adolescente , Adulto Jovem , Insuficiência Pancreática Exócrina , Elastase Pancreática , Fibrose Cística , Genótipo , Estudos TransversaisRESUMO
Abnormalities in lungs caused by emphysema might alter their response to sepsis and the occurrence of acute lung injury (ALI). This study compared the extension of ALI in response to intraperitoneal lipopolysaccharide (LPS) injection in Wistar rats with and without emphysema induced by elastase. Adult male Wistar rats were randomized into four groups: control, emphysema without sepsis, normal lung with sepsis and emphysema with sepsis. Sepsis was induced, and 24 h later the rats were euthanised. The following analysis was performed: blood gas measurements, bronchoalveolar lavage (BAL), lung permeability and histology. Animals that received LPS showed significant increase in a lung injury scoring system, inflammatory cells in bronchoalveolar lavage (BAL) and IL-6, TNF-α and CXCL2 mRNA expression in lung tissue. Animals with emphysema and sepsis showed increased alveolocapillary membrane permeability, demonstrated by higher BAL/serum albumin ratio. In conclusion, the presence of emphysema induced by elastase increases the inflammatory response in the lungs to a systemic stimulus, represented in this model by the intraperitoneal injection of LPS.
Assuntos
Lesão Pulmonar Aguda/patologia , Elastase Pancreática/efeitos adversos , Enfisema Pulmonar/patologia , Síndrome do Desconforto Respiratório/patologia , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/metabolismo , Animais , Líquido da Lavagem Broncoalveolar , Quimiocina CXCL2/genética , Quimiocina CXCL2/metabolismo , Modelos Animais de Doenças , Inflamação/induzido quimicamente , Inflamação/metabolismo , Inflamação/patologia , Injeções Intraperitoneais , Interleucina-6/genética , Interleucina-6/metabolismo , Lipopolissacarídeos/efeitos adversos , Pulmão/metabolismo , Pulmão/patologia , Masculino , Enfisema Pulmonar/induzido quimicamente , Enfisema Pulmonar/metabolismo , Ratos , Ratos Wistar , Síndrome do Desconforto Respiratório/induzido quimicamente , Síndrome do Desconforto Respiratório/metabolismo , Sepse/induzido quimicamente , Sepse/metabolismo , Sepse/patologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Context Fecal elastase is a noninvasive test for pancreatic insufficiency diagnosis. Objectives Evaluate the usefulness of fecal elastase 1 for the indication of exocrine pancreatic insufficiency among former alcohol addicts and patients with chronic pancreatitis. Methods Forty-three patients with chronic pancreatitis and thirty-three asymptomatic former alcohol addicts entered the study. The levels of fecal elastase 1 were measured using a commercial kit. Pancreatic imaging findings were used to categorize the groups. Results The levels of fecal elastase 1 were significantly lower in the patients than in the former alcohol addicts and in the group with tissue calcifications, duct alterations, or atrophy. With a cutoff level of 100 μg/g, the sensitivity of fecal elastase 1 in chronic pancreatitis was 46.51% and its specificity was 87.88% with a positive predictive value of 83.33% and a negative predictive value of 55.77%. When patients were stratified according to the severity of their pancreatitis, the sensitivity was 6.25% for mild pancreatitis and 70.37% for marked pancreatitis. Conclusion Low level of fecal elastase 1 was associated with marked rather than mild chronic pancreatitis; however, it may be useful to indicate pancreatic exocrine insufficiency in asymptomatic former alcohol addicts. .
Contexto O teste de elastase fecal é um teste não invasivo para diagnosticar insuficiência pancreática. Objetivos Avaliar a utilidade da elastase fecal 1 como indicador de insuficiência pancreática entre ex alcoólatras e pacientes com pancreatite crônica. Métodos Quarenta e três pacientes com pancreatite crônica e 33 ex alcoólatras assintomáticos entraram no estudo. Os níveis de elastase fecal 1 foram medidos usando kit comercial. Os achados de imagem pancreática foram usados para categorizar os grupos. Resultados Os níveis de elastase fecal 1 foram significantemente menores nos pacientes que nos ex alcoólatras e no grupo com calcificações teciduais, alterações de ductos, ou atrofia. A sensibilidade da elastase fecal 1 na pancreatite crônica foi de 46,51% e a especificidade foi de 87,88%, com valor preditivo positivo de 83,33% e valor preditivo negativo de 55,77%. Quando os pacientes foram estratificados segundo a severidade da pancreatite, a sensibilidade foi de 6,25% para pancreatite crônica leve e 70,37% para pancreatite crônica severa. Conclusão Baixo nível de elastase fecal foi associado com pancreatite crônica severa mais do que com a leve; entretanto, pode ser útil para indicar insuficiência pancreática exócrina entre os ex alcoólatras. .
Assuntos
Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Alcoolismo/complicações , Insuficiência Pancreática Exócrina/diagnóstico , Fezes/química , Elastase Pancreática/análise , Pancreatite Crônica/complicações , Biomarcadores/análise , Insuficiência Pancreática Exócrina/enzimologia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Índice de Gravidade de DoençaRESUMO
Introduction: Mouse models of emphysema are important tools for testing different therapeutic strategies. The aim of this study was to develop a mouse model of emphysema induced by different doses of elastase in order to produce different degrees of severity. Methods: Thirty female mice (C57BL/6) were used in this study. Different doses of porcine pancreatic elastase were administered intratracheally once a week for four weeks, as follows: 0.1 U (n=8), 0.15 U (n=7), and 0.2 U (n=7). Control mice (n=8) received 50 microL of sterile saline solution intratracheally. Lung mechanics were analyzed by plethysmography. Mean linear intercept and volume fraction occupied by collagen and elastic fibers were determined. Results: An increase in lung resistance was observed with 0.2 U of elastase [median (P-25-P75): 2.02 (1.67; 2.34) cmH2O.s/mL], as well as a decrease in tidal volume and minute ventilation. Peak expiratory flow increased significantly in the groups treated with 0.15 U and 0.2 U of elastase. Mean linear intercept was higher with 0.15 U and 0.2 U of elastase, with destruction of alveolar walls [median (P-25-P75): 30.31 (26.65-43.13) microm and 49.49 (31.67-57.71) microm respectively]. The volume fraction occupied by collagen and elastic fibers was lower in the group receiving 0.2 U of elastase. Conclusion: Four intratracheal instillations of 0.2 U of elastase once a week induced changes in lung function and histology, producing an experimental model of severe pulmonary emphysema, whereas 0.15 U resulted in only histological changes.
Assuntos
Animais , Camundongos , Elastase Pancreática/administração & dosagem , Elastase Pancreática/toxicidade , Enfisema Pulmonar/induzido quimicamente , Enfisema Pulmonar/metabolismoRESUMO
Introdução: A Doença Pulmonar Obstrutiva Crônica (DPOC) está freqüentemente associada a comorbidades crônicas como a doença cardiovascular, o diabetes mellitus e a hipertensão. O presente estudo tem por objetivo investigar as alterações morfológicas e funcionais no coração secundárias ao enfisema pulmonar em ratos diabéticos. Métodos: Ratos Wistar machos adultos (200 ± 20 g, n = 36) foram destinados à avaliação ecocardiográfica, análise morfométrica do coração e pulmões e análise da taxa de sobrevida. O diabetes mellitus foi induzido por aloxana (42 mg/kg, iv) 10 dias antes da indução do enfisema pulmonar por instilação de elastase (0,25 UI/100 g de peso corpóreo). Um grupo de ratos diabéticos recebeu tratamento com insulina NPH (4 UI antes da elastase, seguido de 2 UI/dia, 50 dias). Os experimentos foram realizados 50 dias após a instilação. Resultados: Ratos diabéticos e respectivos controles instilados com elastase apresentaram aumentos similares no diâmetro médio alveolar, cujos valores correlacionam-se positivamente com aumentos na espessura da parede (p=0,0022), na área da cavidade (p=0,0001) e espessura dos cardiomiócitos (p=0,0001) do ventriculo direito (VD). Ratos tornados diabéticos por injeção de aloxana exibiram redução na espessura da parede do ventrículo esquerdo (VE), no septo interventricular (IV) e na espessura dos cardiomiócitos. Estas variáveis morfométricas associaram-se à redução da fração de encurtamento do VE (p < 0,05) e a aumento no tempo de relaxamento isovolumétrico do VE (p < 0,05). A taxa de sobrevida reduziu-se de 80% em ratos diabéticos a 40% em ratos diabéticos instilados com elastase (p < 0,05). Conclusões: O diabetes por aloxana em ratos não modifica a hipertrofia do VD secundária ao enfisema pulmonar, porém induz disfunção ventricular esquerda. A manifestação de ambas as doenças, diabetes mellitus e enfisema pulmonar, reduz substancialmente a taxa de sobrevida, enfatizando a condição de comorbidade na coexistência de...
Background: Chronic Obstructive Pulmonary Disease (COPD) is often associated with chronic comorbid conditions of cardiovascular disease, diabetes mellitus and hypertension. This study aimed to investigate morphological and functional alterations of the heart secondary to chronic emphysema in diabetic rats. Methods: Adult male Wistar rats (200 ± 20 g, n=36) were used for echocardiographic measurements, morphometric analyses of the heart and lungs, and survival rate. Diabetes mellitus was induced by alloxan (42 mg/kg, iv) 10 days before the induction of pulmonary emphysema by the instillation of elastase (0.25 IU/100 g body weight). A group of diabetic rats was treated with NPH insulin (4 IU before elastase, plus 2 IU/day, 50 days). Experiments were performed 50 days after instillation. Results: Both elastase-instilled diabetic rats and matching controls exhibited similar increases in mean alveolar diameter, which are positively correlated with increases in RV wall thickness (p=0.0022), cavity area (p=0.0001), and cardiomyocyte thickness (p=0.0001). Alloxan-diabetic rats demonstrated a reduction in left ventricular (LV) wall, IV septum, and cardiomyocyte thickness, associated with a reduction in LV fractional shortening (p<0.05), and an increase in LViv relaxation time (p < 0.05). Survival rate decreased from 80% in diabetic rats to 40% in elastase-instilled diabetic rats. Conclusions: Alloxan diabetes did not affect RV hypertrophy secondary to chronic emphysema, but induced LV dysfunction. The association of diabetes and emphysema substantially reduced the survival rate, emphasizing the comorbid condition of the coexistence of diabetes and COPD.