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1.
Mol Immunol ; 120: 179-186, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32169738

RESUMO

BACKGROUND: The NLRP3 inflammasome has been suggested to play a crucial role in host antiviral defense, including against hepatitis B virus (HBV) infection. In the present study, we measured expression of NLRP3 and its related cytokines in patients with different stages of HBV-related acute-on-chronic liver failure (HBV-ACLF), a pattern of end-stage liver disease that occurs frequently in patients with chronic HBV (CHB) infection or HBV-related cirrhosis. METHODS: A total of 75 subjects including 30 HBV-ACLF patients, 30 CHB patients, and 15 healthy controls (HCs) were enrolled. The NLRP3 inflammasome and its components (caspase-1, interleukin (IL)-1ß, and IL-18) were measured in peripheral blood mononuclear cells (PBMCs), macrophages, and liver using flow cytometry, quantitative real-time polymerase chain reaction (RT-PCR), western blot, and immunohistochemistry. The LPS was used to evaluate changes in NLRP3 and its related cytokines in CD14+ monocytes which may reflect immune status. Cytokine expression was measured using RT-PCR. RESULTS: Patients with HBV-ACLF had lower NLRP3 inflammasome expression in peripheral CD14+ monocytes, particularly in the middle-to-late stage, but higher expression in liver macrophages compared to CHB and HCs. Compared with H-LPS or L-LPS alone, L-LPS sequential H-LPS can significantly inhibit the expression of NLRP3 and its related cytokines. CONCLUSION: Differential expression patterns of the NLRP3 inflammasome in the periphery and liver might be related to immune dysfunction and recruitment of monocytes to the injured liver during disease progression. Persistent systemic inflammation is likely a cause of compromised immune status in patients with HBV-ACLF.


Assuntos
Insuficiência Hepática Crônica Agudizada/imunologia , Hepatite B Crônica/imunologia , Inflamassomos/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , Insuficiência Hepática Crônica Agudizada/etiologia , Insuficiência Hepática Crônica Agudizada/metabolismo , Adulto , Biomarcadores/sangue , Biomarcadores/metabolismo , Estudos de Casos e Controles , Citocinas/sangue , Citocinas/genética , Citocinas/metabolismo , Feminino , Hepatite B Crônica/complicações , Hepatite B Crônica/metabolismo , Humanos , Inflamassomos/sangue , Inflamassomos/metabolismo , Fígado/imunologia , Fígado/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Monócitos/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo
2.
J Bras Pneumol ; 45(4): e20190001, 2019 Aug 29.
Artigo em Português, Inglês | MEDLINE | ID: mdl-31482943

RESUMO

OBJECTIVE: This study aimed to determine the serum levels of NACHT, Leucine-rich repeat (LRR), and Pyrin (PYD) domains-containing Protein 3 (NLRP3) and cathelicidin LL-37, and investigate their prognostic significance in community-acquired pneumonia (CAP). METHODS: The sample of this prospective study was composed of 76 consecutive patients with CAP. Demographic data and clinical characteristics were collected. Serum levels of NLRP3 and LL-37 were determined by ELISA. Spearman's analysis was used to evaluate the correlation between NLRP3 and LL-37. Association of NLRP3 and LL-37 with 30-day survival and mortality rates was assessed using the Kaplan-Meier curve and logistic regression analysis. RESULTS: Serum NLRP3 significantly increased whereas serum LL-37 significantly decreased in patients with severe CAP. Significant correlation was observed between serum NLRP3 and LL-37 in CAP patients. Patients with higher levels of NLRP3 and lower levels of LL-37 showed lower 30-day survival rate and higher mortality compared with those with lower NLRP3 and higher LL-37 levels. CONCLUSION: Severe CAP patients tend to present higher serum NLRP3 and lower serum LL-37, which might serve as potential biomarkers for CAP prognosis.


Assuntos
Peptídeos Catiônicos Antimicrobianos/sangue , Infecções Comunitárias Adquiridas/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Pneumonia/sangue , Proteínas/análise , Pirina/sangue , Idoso , Biomarcadores/sangue , Estudos de Casos e Controles , Infecções Comunitárias Adquiridas/mortalidade , Feminino , Humanos , Estimativa de Kaplan-Meier , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Pneumonia/mortalidade , Valor Preditivo dos Testes , Estudos Prospectivos , Valores de Referência , Medição de Risco , Fatores de Risco , Fatores de Tempo
3.
Vasc Health Risk Manag ; 15: 209-220, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31371977

RESUMO

Cholesterol-embolization syndrome (CES) is a multisystemic disease with various clinical manifestations. CES is caused by embolization of cholesterol crystals (CCs) from atherosclerotic plaques located in the major arteries, and is induced mostly iatrogenically by interventional and surgical procedures; however, it may also occur spontaneously. Embolized CCs lead to both ischemic and inflammatory damage to the target organ. Therefore, anti-inflammatory agents, such as corticosteroids and cyclophosphamide, have been investigated as treatment for CES in several studies, with conflicting results. Recent research has revealed that CES is actually a kind of autoinflammatory disease in which inflammasome pathways, such as NLRP3 and IL1, are induced by CCs. These recent findings may have clinical implications such that colchicine and IL1 inhibitors, namely canakinumab, may be beneficial in the early stages of CES.


Assuntos
Aterosclerose , Colesterol/sangue , Embolia de Colesterol , Corticosteroides/uso terapêutico , Animais , Anti-Inflamatórios/uso terapêutico , Aterosclerose/sangue , Aterosclerose/diagnóstico , Aterosclerose/tratamento farmacológico , Aterosclerose/epidemiologia , Biomarcadores/sangue , Cristalização , Embolia de Colesterol/sangue , Embolia de Colesterol/diagnóstico , Embolia de Colesterol/epidemiologia , Embolia de Colesterol/prevenção & controle , Humanos , Imunossupressores/uso terapêutico , Inflamassomos/sangue , Mediadores da Inflamação/sangue , Interleucina-1/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Placa Aterosclerótica , Prognóstico , Fatores de Risco , Síndrome
4.
Clinics (Sao Paulo) ; 74: e729, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31411276

RESUMO

OBJECTIVES: To investigate the relationship between the serum levels of NLRP3 and HMGB-1 and the prognosis of patients with severe blunt abdominal trauma. METHODS: In total, 299 patients were included in the current study from July 2014 to December 2015. All patients were divided into the mild/moderate blunt abdominal trauma group and the severe blunt abdominal trauma group according to their injury severity scores. Serum levels of NLRP3 and HMGB-1 were measured upon admission (0 h) and at 12 h, 24 h, 48 h, 72 h and 7 days after admission. RESULTS: Compared with the healthy controls, both the mild/moderate and severe blunt abdominal trauma groups had higher serum levels of NLRP3 and HMGB-1 at admission. At all points, the serum levels of NLRP3 and HMGB-1 were significantly higher in the severe group than in the mild/moderate group. The serum levels of both NLRP3 and HMGB-1 were significantly higher in the deceased patients than in the living patients. The Kaplan-Meier curve showed that compared with patients with higher levels of NLRP3 or HMGB-1, those with lower levels had longer survival times. The serum levels of both NLRP3 and HMGB-1 were independent risk factors for 6-month mortality in severe blunt abdominal trauma patients. CONCLUSION: The serum levels of NLRP3 and HMGB-1 were significantly elevated in severe blunt abdominal trauma patients, and the serum levels of both NLRP3 and HMGB-1 were correlated with 6-month mortality in severe blunt abdominal trauma patients.


Assuntos
Traumatismos Abdominais/sangue , Proteína HMGB1/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Traumatismos Abdominais/mortalidade , Traumatismos Abdominais/terapia , Adulto , China/epidemiologia , Feminino , Humanos , Escala de Gravidade do Ferimento , Masculino , Pessoa de Meia-Idade , Prognóstico
5.
Mol Med Rep ; 20(2): 1826-1836, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31257469

RESUMO

Nucleotide­binding oligomerization domain, leucine rich repeat, and pyrin domain­containing protein 3 (NLRP3) inflammasome has been implicated in a series of physiological and pathological processes. However, its correlation in coronary heart disease (CHD) still remains to be elucidated. The present study aimed to determine the expression of NLRP3 inflammasome in peripheral blood monocytes (PBMCs) of stable angina pectoris (SAP) and acute myocardial infarction (AMI) patients. In addition, the effect of rosuvastatin on their activities was analyzed in vitro. A total of 60 participants with SAP (n=20), AMI (n=20) and non­CHD controls (n=20) were enrolled. Fluorescence­activated cell sorting, real­time PCR, western blotting and enzyme­linked immunosorbent assay were performed to reveal the role of NLRP3 inflammasome. NLRP3 inflammasome was expressed in the PBMCs, and revealed an increased expression along the downstream interleukin (IL)­1ß and IL­18 in both SAP and AMI groups, compared to the control group. Moreover, there was a more marked increase in the expression of these indicators in AMI patients when compared to SAP patients. Interference with rosuvastatin in vitro revealed that the expression of NLRP3 inflammasome and its downstream cytokines were significantly downregulated in both SAP and AMI groups in a time­dependent manner. The activation of NLRP3 inflammasome may be involved in the development of CHD, and rosuvastatin could attenuate the inflammatory process of atherosclerosis by downregulating NLRP3 expression and its downstream mediators. These findings indicated a potential role of NLRP3 in the pathogenesis and management of CHD, and also provided new insights into the mechanistic framework of rosuvastatin activity.


Assuntos
Angina Estável/sangue , Doença das Coronárias/sangue , Infarto do Miocárdio/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Rosuvastatina Cálcica/administração & dosagem , Idoso , Angina Estável/tratamento farmacológico , Angina Estável/patologia , Doença das Coronárias/tratamento farmacológico , Doença das Coronárias/patologia , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Inflamassomos/efeitos dos fármacos , Inflamassomos/genética , Inflamação/sangue , Inflamação/tratamento farmacológico , Inflamação/patologia , Masculino , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/patologia , Transdução de Sinais/efeitos dos fármacos
6.
Diab Vasc Dis Res ; 16(4): 360-368, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30939905

RESUMO

BACKGROUND: Adipose tissue produces pro-inflammatory mediators involved in the atherosclerotic process. We investigated whether 12-month exercise training in patients with type 2 diabetes mellitus and coronary artery disease would reduce circulating levels and genetic expression of mediators in the interleukin-18, Caspase-1 and NLR pyrin domain containing 3 pathways. Correlations to glucometabolic variables; fasting glucose, HbA1c, duration of diabetes, insulin, C-peptide, insulin resistance (measured by homeostatic model assessment indexes - insulin resistance) and body mass index at baseline were further assessed. METHODS: 137 patients (aged 41-81 years, 17.2% female participants) were included and randomized to a 12-month exercise programme or to a control group. Fasting blood and adipose tissue samples were taken at inclusion and after 12 months. RESULTS: No statistically significant difference in changes of any variable between the intervention and the control group was found. At baseline, a positive correlation between insulin and homeostatic model assessment indexes - insulin resistance, interleukin-18 expression in adipose tissue and an inverse correlation between some glucometabolic variables and leukocyte expression of NLR pyrin domain containing 3 and Caspase-1 were observed. CONCLUSION: No significant effects of long-term exercise training were observed on the inflammasome-related mediators in our patients with combined coronary artery disease and type 2 diabetes mellitus. The observed correlations may indicate a pro-inflammatory state in adipose tissue by overweight and a compensatory downregulation of these mediators in circulating leucocytes.


Assuntos
Glicemia/metabolismo , Doença da Artéria Coronariana/terapia , Diabetes Mellitus Tipo 2/terapia , Terapia por Exercício , Inflamassomos/sangue , Mediadores da Inflamação/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Índice de Massa Corporal , Caspase 1/sangue , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/diagnóstico , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Feminino , Hemoglobina A Glicada/metabolismo , Humanos , Insulina/sangue , Interleucina-18/sangue , Masculino , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Fatores de Tempo , Resultado do Tratamento
7.
Int J Med Sci ; 16(3): 461-469, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30911280

RESUMO

Acute-on-chronic hepatitis B liver failure (ACHBLF) refers to the acute deterioration of liver function during chronic hepatitis B virus infection, and is associated with high mortality, with rapid progression to death. Nucleotide-binding oligomerisation domain-like receptors (NLRs) Family Pyrin Domain Containing 3(NLRP3) inflammasome contributed to the pathogenesis of D-galactosamine and lipopolysaccharide-induced acute liver failure. However, the profile of NLRP3 in patients with ACHBLF has not been demonstrated. This study was therefore conducted to investigate the expression of NLRP3 in patients with ACHBLF and identify the effect of glucocorticoid on NLRP3. We recruited 70 patients with ACHBLF undergoing glucocorticoid treatment for 28 days, 30 patients with chronic hepatitis B (CHB), and 24 healthy controls (HCs) in this study. The relative messenger RNA (mRNA) level of NLRP3 and related genes were measured by reverse transcription polymerase chain reaction, the plasma levels of interleukin-1ß (IL-1ß) and interleukin-18 (IL-18) were measured by enzyme-linked immunosorbent assay. The mRNA level of NLRP3 was significantly higher in patients with ACHBLF than in patients with CHB as well as HCs (P<0.05). The plasma levels of IL-1ß and IL-18 in patients with ACHBLF were significantly higher than in patients with CHB and HCs (P<0.05). The relative mRNA level of NLRP3 in surviving patients decreased significantly compared with that in patients who did not survive after glucocorticoid treatment (P<0.05). In conclusion, NLRP3 increased in patients with ACHBLF. Glucocorticoid could downregulate the expression of NLRP3 in surviving patients with ACHBLF.


Assuntos
Insuficiência Hepática Crônica Agudizada/etiologia , Glucocorticoides/uso terapêutico , Hepatite B Crônica/complicações , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Insuficiência Hepática Crônica Agudizada/tratamento farmacológico , Insuficiência Hepática Crônica Agudizada/mortalidade , Adulto , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Hepatite B Crônica/tratamento farmacológico , Hepatite B Crônica/metabolismo , Humanos , Interleucina-18/sangue , Interleucina-1beta/sangue , Leucócitos Mononucleares/metabolismo , Masculino , Metilprednisolona/uso terapêutico , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Prednisolona/uso terapêutico
8.
Biochem Biophys Res Commun ; 508(2): 614-619, 2019 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-30522866

RESUMO

Platelets play a critical role in the pathophysiology of peripheral arterial disease (PAD). The mechanisms by which muscle ischemia regulates aggregation of platelets are poorly understood. We have recently identified the Nod-like receptor nucleotide-binding domain leucine rich repeat containing protein 3 (NLRP3) expressed by platelets as a critical regulator of platelet activation and aggregation, which may be triggered by activation of toll-like receptor 4 (TLR4). In this study, we performed femoral artery ligation (FAL) in transgenic mice with platelet-specific ablation of TLR4 (TLR4 PF4) and in NLRP3 knockout (NLRP3-/-) mice. NLRP3 inflammasome activity of circulating platelets, as monitored by activation of caspase-1 and cleavage of interleukin-1ß (IL-1ß), was upregulated in mice subjected to FAL. Genetic ablation of TLR4 in platelets led to decreased platelet caspase 1 activation and platelet aggregation, which was reversed by the NLRP3 activator Nigericin. Two weeks after the induction of FAL, ischemic limb perfusion was increased in TLR4 PF4 and NLRP3-/- mice as compared to control mice. Hence, activation of platelet TLR4/NLRP3 signaling plays a critical role in upregulating platelet aggregation and interfering with perfusion recovery in muscle ischemia and may represent a therapeutic target to improve limb salvage.


Assuntos
Modelos Animais de Doenças , Membro Posterior/metabolismo , Inflamassomos/metabolismo , Isquemia/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Receptor 4 Toll-Like/metabolismo , Animais , Inflamassomos/sangue , Isquemia/sangue , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/deficiência , Ativação Plaquetária , Agregação Plaquetária , Transdução de Sinais , Receptor 4 Toll-Like/sangue , Regulação para Cima
9.
J. bras. pneumol ; 45(4): e20190001, 2019. tab, graf
Artigo em Português | LILACS | ID: biblio-1019982

RESUMO

RESUMO Objetivo Este estudo teve como objetivo determinar os níveis séricos de proteína 3 contendo um domínio NACHT, porção C-terminal rica em repetições de leucina e de domínio pirina (NLRP3) e catelicidina LL-37, bem como investigar sua importância prognóstica em pneumonia adquirida na comunidade (PAC). Métodos Este estudo prospectivo incluiu 76 pacientes com PAC. Foram obtidos dados demográficos e características clínicas. Os níveis séricos de NLRP3 e LL-37 foram determinados por meio do teste ELISA. A correlação entre NLRP3 e LL-37 foi estimada por intermédio da análise de Spearman. A associação entre NLRP3 e LL-37 com 30 dias de taxa de sobrevida e de mortalidade foi avaliada pela curva de Kaplan-Meier e análise de regressão logística. Resultados Os níveis séricos de NLRP3 estavam elevados, enquanto os níveis de LL-37 apresentaram redução significativa em pacientes com PAC grave. Observou-se correlação significativa entre os níveis séricos de NLRP3 e LL-37 em pacientes com PAC. Pacientes com níveis elevados de NLRP3 e níveis reduzidos de LL-37 exibiram maior taxa de sobrevida em 30 dias e de mortalidade quando comparados com aqueles com níveis inferiores de NLRP3 e LL-37. Conclusões Pacientes com PAC grave tendem a apresentar níveis séricos elevados de NLRP3 e níveis reduzidos de LL-37, o que pode ser utilizado como um potencial biomarcador prognóstico.


ABSTRACT Objective This study aimed to determine the serum levels of NACHT, Leucine-rich repeat (LRR), and Pyrin (PYD) domains-containing Protein 3 (NLRP3) and cathelicidin LL-37, and investigate their prognostic significance in community-acquired pneumonia (CAP). Methods The sample of this prospective study was composed of 76 consecutive patients with CAP. Demographic data and clinical characteristics were collected. Serum levels of NLRP3 and LL-37 were determined by ELISA. Spearman's analysis was used to evaluate the correlation between NLRP3 and LL-37. Association of NLRP3 and LL-37 with 30-day survival and mortality rates was assessed using the Kaplan-Meier curve and logistic regression analysis. Results Serum NLRP3 significantly increased whereas serum LL-37 significantly decreased in patients with severe CAP. Significant correlation was observed between serum NLRP3 and LL-37 in CAP patients. Patients with higher levels of NLRP3 and lower levels of LL-37 showed lower 30-day survival rate and higher mortality compared with those with lower NLRP3 and higher LL-37 levels. Conclusion Severe CAP patients tend to present higher serum NLRP3 and lower serum LL-37, which might serve as potential biomarkers for CAP prognosis.


Assuntos
Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Idoso , Pneumonia/sangue , Proteínas/análise , Infecções Comunitárias Adquiridas/sangue , Peptídeos Catiônicos Antimicrobianos/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Pirina/sangue , Pneumonia/mortalidade , Biomarcadores/sangue , Estudos de Casos e Controles , Modelos Logísticos , Análise Multivariada , Valor Preditivo dos Testes , Estudos Prospectivos , Infecções Comunitárias Adquiridas/mortalidade , Estimativa de Kaplan-Meier
10.
J Cardiovasc Pharmacol ; 72(6): 303-307, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30422890

RESUMO

BACKGROUND: A diet rich in saturated fat and sugars (Western diet, WD) induces myocardial expression of the NLRP3 inflammasome and dysfunction in mice. We therefore hypothesized that a diet enriched with an orally available NLRP3 inflammasome inhibitor could prevent WD-induced cardiac dysfunction in mice. METHODS: Ten-week-old CD-1 male mice were fed WD or standard diet (SD) for 8 weeks. The compound 16673-34-0, an orally active NLRP3 inhibitor, was added to the diet at a concentration of 100 mg/Kg. The plasmatic levels of the NLRP3 inflammasome inhibitor were measured. Food intake, body weight, and glucose tolerance were assessed. Cardiac systolic and diastolic functions were measured by Doppler echocardiography at baseline, 4 weeks, and 8 weeks. RESULTS: WD induced a significant increase in body weight (+14%, P = 0.02), impaired glucose tolerance (+34%, P = 0.03), and a significant increase in isovolumetric relaxation time (+129%, P = 0.03) and reduction in left ventricular ejection fraction (-10%, P = 0.03), as compared to standard chow diet (SD). The treatment with NLRP3 inhibitor in the diet prevented cardiac systolic and diastolic dysfunction (P < 0.05 for left ventricular ejection fraction, isovolumetric relaxation time, and myocardial performance index in WD with drug vs. WD without drug), without significant changes in heart rate and metabolic parameters. CONCLUSIONS: An orally available NLRP3 inhibitor prevented WD-induced cardiac dysfunction in obese mice.


Assuntos
Anti-Inflamatórios/administração & dosagem , Dieta Ocidental , Inflamassomos/antagonistas & inibidores , Proteína 3 que Contém Domínio de Pirina da Família NLR/antagonistas & inibidores , Obesidade/tratamento farmacológico , Volume Sistólico/efeitos dos fármacos , Disfunção Ventricular Esquerda/prevenção & controle , Função Ventricular Esquerda/efeitos dos fármacos , Administração Oral , Animais , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Diástole , Modelos Animais de Doenças , Ecocardiografia Doppler , Inflamassomos/sangue , Interleucina-18/sangue , Masculino , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Obesidade/sangue , Obesidade/etiologia , Obesidade/fisiopatologia , Sístole , Fatores de Tempo , Disfunção Ventricular Esquerda/sangue , Disfunção Ventricular Esquerda/etiologia , Disfunção Ventricular Esquerda/fisiopatologia
11.
J Complement Integr Med ; 16(2)2018 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-30335608

RESUMO

Background Chronic pancreatitis (CP) is a persistent inflammation of the pancreas clinically presented with severe abdominal pain, progressive fibrosis, and loss of exocrine and endocrine functions. Inflammasomes, cytosolic multiprotein complexes which regulate the formation of proinflammatory cytokines, are influenced by various factors including heat shock proteins (HSPs). Morus alba L., or white mulberry root bark is a valued traditional Asian medicine with a diverse array of phytochemicals. The aim of this investigation was to define the modulatory action of methanolic extract of Morus alba root bark (MEMARB) on NLRP3 inflammasome, and HSPs in pancreas subjected to inflammatory insult. Methods Pancreatitis was induced in male albino Wistar rats by ethanol (0-36%) and cerulein (20 µg/kg b.wt., i.p.) for 5 weeks with or without MEMARB administration. Serum lipase/amylase (L/A) ratio, oxidative stress index (OSI) and reduced glutathione (GSH)/oxidized glutathione (GSSG) ratio in the pancreas were evaluated. Levels of serum HSP70 was quantified by ELISA. NF-kappa B, NLRP3-ASC, caspase-1, IL-1ß, IL-18, and HSP70 gene expression was quantified by quantitative real-time polymerase chain reaction (qPCR). Results L/A ratio and oxidative stress determined in terms of OSI and GSH/GSSG ratio were elevated in pancreatitis-induced rats. The levels were restored in MEMARB co-administered animals. Serum level of HSP70 was increased in pancreatitis-induced animals and dropped significantly in MEMARB co-administrated rats. Pancreatitis-induced group showed increased expression of NF-kappa B, IL-1ß, IL-18, caspase-1, NLRP3-ASC and HSP70 mRNA than in MEMARB treated group. Conclusions It can be concluded that the M. alba root extract modulates the expression of HSP70 and NLRP3-ASC which might be attributed to its pancreato-protective effect.


Assuntos
Proteínas de Choque Térmico HSP70/genética , Morus/química , Pancreatite/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Animais , Biomarcadores/sangue , Ceruletídeo/efeitos adversos , Etanol/efeitos adversos , Proteínas de Choque Térmico HSP70/sangue , Humanos , Inflamassomos/sangue , Inflamassomos/genética , Interleucina-18/sangue , Interleucina-18/genética , Interleucina-1beta/sangue , Interleucina-1beta/genética , Masculino , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Pancreatite/sangue , Pancreatite/induzido quimicamente , Pancreatite/genética , Casca de Planta/química , Raízes de Plantas/química , Ratos , Ratos Wistar
12.
Clin Chim Acta ; 486: 269-274, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30138619

RESUMO

OBJECTIVE: There is some evidence that an enhanced Th17 response is attributable to interleukin-1ß (IL-1ß) matured by activation of the NLRP3 inflammasome. In this study, we assessed the expression of NLRP3 inflammasome components and their associated proinflammatory cytokines in patients with ankylosing spondylitis (AS). METHODS: A total of 23 male patients with AS and 29 male controls were recruited and peripheral blood mononuclear cells (PBMCs) were collected. Transcript and protein expression of NLRP3, caspase-1, ASC, IL-1ß, IL-17A, and IL-23 were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assay, respectively. Data were analyzed using Spearman's correlation coefficient, Mann-Whitney U test, and receiver operating characteristic curves. RESULTS: Higher mRNA expression of NLRP3, ASC, IL-1ß, IL-17A, and IL-23 was noted in the PBMCs of AS patients than those of controls (p = 0.013, p = 0.001, p = 0.002, p = 0.011, and p = 0.037, respectively). Similarly, protein expression of NLRP3, caspase-1, ASC, IL-1ß, IL-17A, and IL-23 was higher in the AS group than the control group. There were significant correlations among NLRP3, caspase-1, ASC, IL-1ß, IL-17A, and IL-23 expression in patients with AS, except for a weak association between NLRP3 and IL-17A. Treatment of cultured PBMCs with 10 ng of lipopolysaccharide induced NLRP3, caspase-1, ASC, IL-1ß, IL-17A, and IL-23 expression, which was marked suppressed by treatment with ascorbic acid. CONCLUSION: This study suggests that the NLRP3 inflammasome may be involved in the pathogenesis of AS and therefore a potential therapeutic target in AS.


Assuntos
Citocinas/sangue , Inflamassomos/sangue , Leucócitos Mononucleares/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Espondilite Anquilosante/sangue , Adulto , Western Blotting , Citocinas/genética , Voluntários Saudáveis , Humanos , Inflamassomos/genética , Leucócitos Mononucleares/patologia , Masculino , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , RNA Mensageiro/sangue , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Espondilite Anquilosante/genética , Espondilite Anquilosante/patologia
14.
Invest Ophthalmol Vis Sci ; 59(2): 978-985, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29450537

RESUMO

Purpose: The aim of this study was to determine the association between nucleotide-binding and oligomerization domain (NOD)-like receptors (NLRs) family, pyrin domain-containing 3 (NLRP3) inflammasome-induced inflammation and disease severity in diabetic retinopathy (DR). Methods: Blood samples were collected from 64 patients with diabetes (DR, 43; without DR, 21) and 25 healthy controls. The protein and mRNA expression levels of NLRP3 inflammasomes in peripheral blood mononuclear cells were determined using western blotting and quantitative real-time reverse transcription-PCR. A total of 82 vitreous samples were obtained from patients with DR (n = 60) and nondiabetic controls (n = 22). All patients were candidates for vitrectomy. Interleukin (IL)-1ß and IL-18 in the peripheral blood mononuclear cell culture medium and vitreous fluid were detected by enzyme-linked immunosorbent assay (ELISA). Immunofluorescence staining for apoptosis-associated speck-like protein with a caspase recruitment domain (ASC) and NLRP3 was performed in fibrovascular membranes from 21 proliferative DR patients and 22 controls with idiopathic epiretinal membranes. Results: We observed increased gene and protein expression of NLRP3, ASC, and caspase-1 in peripheral blood mononuclear cells of adults with DR compared with that in normal controls. Furthermore, the elevated expressions of NLRP3 and ASC were observed in the fibrovascular membranes from 21 adults with proliferative DR when compared with the 22 controls. IL-1ß and IL-18 in the peripheral blood mononuclear cells and vitreous fluid were elevated in the DR patients when compared with controls. Conclusions: These outcomes suggested that NLRP3 inflammasomes are upregulated in adults with DR and may play a key role in the pathogenesis and progression of DR.


Assuntos
Retinopatia Diabética/metabolismo , Regulação da Expressão Gênica/fisiologia , Inflamassomos/metabolismo , Inflamação/metabolismo , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Western Blotting , Proteínas Adaptadoras de Sinalização CARD/genética , Caspase 1/genética , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Interleucina-18/sangue , Interleucina-1beta/sangue , Leucócitos Mononucleares/metabolismo , Masculino , Pessoa de Meia-Idade , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , RNA Mensageiro/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa
15.
Clin Immunol ; 191: 100-109, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29183866

RESUMO

The proinflammatory protease caspase-1 plays pivotal roles in central pathways of innate immunity, thereby contributing to pathogen clearance. Beside its physiological role, dysregulated activity of caspase-1 is known to contribute to an increasing number of diseases. In this study, we optimized and validated a low-volume human whole blood assay facilitating the measurement of caspase-1 activation and inflammasome-related gene expression upon stimulation of the NLRP3, NLRC4 or AIM2 inflammasome. Using the NLRP3 inflammasome specific inhibitor MCC950, we were able to measure the activity of canonical or alternative NLRP3 pathways, AIM2 and NLRC4 inflammasomes in whole blood. Based on our data we assume a superposition of NLRP3 and NLRC4 inflammasome activities in human whole blood following stimulation with S. typhimurium. The optimized whole blood assay may be suitable for diagnostic and research purposes for pediatric patients who can only donate small amounts of blood.


Assuntos
Proteínas Adaptadoras de Sinalização CARD/sangue , Proteínas de Ligação ao Cálcio/sangue , Proteínas de Ligação a DNA/sangue , Inflamassomos/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Coleta de Amostras Sanguíneas , Caspase 1/fisiologia , Humanos , Interleucina-1beta/fisiologia , Salmonella typhimurium
17.
Georgian Med News ; (268-269): 12-17, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28820405

RESUMO

After discovery of antiphospholipid syndrome (APS) our understanding of molecular mechanisms of living matter has become more sophisticated and on this way monocytes has become crucial player, particularly in pathogenesis of APS. Thrombotic and non-thrombotic complications of APS could be explained by monocytes' activation too. But mechanisms underlying their activation are poorly investigated. So we aimed to determine transcriptional activity of monocytes after exposing them to low concentrations of lipopolysaccharide (LPS) and LPS+ATP using comparative of RT-PCR. Our study included eleven women suffering from recurrent miscarriages and APS (mean age 30±5,6 years). Nine healthy women (mean age of 29±8,5 years) without a positive family history of APS, autoimmune diseases and thrombosis were chosen as a control group. The results showed increasing levels of TLR2, IL-23, CCL2, CXCL10, IL-1ß and IL-6 in APS cells, while in healthy cells LPS resulted in IL-6 and STAT3 elevated mRNAs. Double stimulation of APS cells resulted in decreased mRNA levels of CCL-2, IL-1ß, and mRNA NLRP3 in healthy cells. At the same time TLR2 mRNAs were elevated in both groups after double stimulation. Thus increased sensitivity of APS cells to LPS may contribute to thrombus formation. Low concentration of ATP diminishes LPS-induced inflammatory state of APS monocytes, which might be one of potential regulatory mechanisms.


Assuntos
Síndrome Antifosfolipídica/metabolismo , Monócitos/metabolismo , Aborto Habitual/imunologia , Trifosfato de Adenosina/farmacologia , Adulto , Síndrome Antifosfolipídica/imunologia , Síndrome Antifosfolipídica/patologia , Estudos de Casos e Controles , Quimiocinas/sangue , Quimiocinas/genética , Feminino , Expressão Gênica , Humanos , Técnicas In Vitro , Interleucinas/sangue , Interleucinas/genética , Lipopolissacarídeos/farmacologia , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , RNA Mensageiro/metabolismo , Receptor 2 Toll-Like/sangue , Receptor 2 Toll-Like/genética , Adulto Jovem
18.
IUBMB Life ; 69(8): 623-630, 2017 08.
Artigo em Inglês | MEDLINE | ID: mdl-28631886

RESUMO

Diabetic nephropathy (DN) is one of the major causes of end-stage renal disease. Nod-like receptors nucleotide-binding domain and leucine-rich repeat pyrin-3 domain (NLRP3) inflammasome displays a considerable role in the chronic inflammatory state observed in diabetic patients. Urinary heat shock protein 72 (uHSP72) is a sensitive and specific biomarker for the early detection of acute kidney injury. The aim of this study was to evaluate NLRP3 relative gene expression, its correlation with inflammatory and oxidative stress markers, and to assess the value of uHSP72 in the early detection of DN in type 2 diabetic patients with different degrees of DN. Forty-five type 2 diabetic patients: 15 normoalbuminuric, 15 microalbuminuric, 15 macroalbuminuric, in addition to 15 healthy controls were enrolled in this study. Clinical examination and routine laboratory investigations were performed. NLRP3 mRNA expression was assessed by real time polymerase chain reaction. Serum 8-hydroxy-2'-deoxyguanosine (8-OHdG), interleukin 1ß (IL-1ß), and uHSP72 levels were estimated by enzyme-linked immunosorbent assay. Serum chitotriosidase (CHIT1) activity was examined. NLRP3 mRNA relative expression, serum levels of 8-OHdG, IL-1ß, and uHSP72, in addition to CHIT 1 activity were significantly increased in the macroalbuminuric patient group as compared to control and the other two diabetic groups. Also, a significant positive correlation was documented between the previously mentioned parameters and urinary albumin/creatinine ratio, serum creatinine, and HbA1c. Multiple linear regression analysis using urinary albumin/creatinine ratio as dependent variable confirmed that uHSP72 and NLRP3 mRNA relative expression were the independent predictors of DN (ß were 0.432 and 0.448 respectively, P < 0.001). Receiver operating characteristic analyses revealed that both NLRP3 mRNA relative expression and uHSP72 levels were useful biomarkers discriminating DN patients from patients with type 2 diabetes mellitus (AUC were 0.957 and 0.983, respectively). uHSP72 may be considered as a novel potential diagnostic biomarker for the early detection of DN. Moreover, these data support the pivotal role of NLRP3 in the development and progression of DN. © 2017 IUBMB Life, 69(8):623-630, 2017.


Assuntos
Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/urina , Proteínas de Choque Térmico HSP72/urina , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Adulto , Biomarcadores/sangue , Biomarcadores/urina , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/urina , Nefropatias Diabéticas/etiologia , Nefropatias Diabéticas/patologia , Feminino , Humanos , Inflamassomos/sangue , Inflamassomos/urina , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/genética
19.
Crit Care ; 20(1): 170, 2016 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-27260481

RESUMO

BACKGROUND: Whole body ischemia-reperfusion injury (IRI) after cardiopulmonary resuscitation (CPR) induces a generalized inflammatory response which contributes to the development of post-cardiac arrest syndrome (PCAS). Recently, pattern recognition receptors (PRRs), such as toll-like receptors (TLRs) and inflammasomes, have been shown to mediate the inflammatory response in IRI. In this study we investigated monocyte PRR signaling and function in PCAS. METHODS: Blood samples were drawn in the first 12 hours, and at 24 and 48 hours following return of spontaneous circulation in 51 survivors after cardiac arrest. Monocyte mRNA levels of TLR2, TLR4, interleukin-1 receptor-associated kinase (IRAK)3, IRAK4, NLR family pyrin domain containing (NLRP)1, NLRP3, AIM2, PYCARD, CASP1, and IL1B were determined by real-time quantitative PCR. Ex vivo cytokine production in response to stimulation with TLR ligands Pam3CSK4 and lipopolysaccharide (LPS) was assessed in both whole blood and monocyte culture assays. Ex vivo cytokine production of peripheral blood mononuclear cells (PBMCs) from a healthy volunteer in response to stimulation with patients' sera with or without LPS was assessed. The results were compared to 19 hemodynamically stable patients with coronary artery disease. RESULTS: Monocyte TLR2, TLR4, IRAK3, IRAK4, NLRP3, PYCARD and IL1B were initially upregulated in patients following cardiac arrest. The NLRP1 and AIM2 inflammasomes were downregulated in resuscitated patients. There was a significant positive correlation between TLR2, TLR4, IRAK3 and IRAK4 expression and the degree of ischemia as assessed by serum lactate levels and the time until return of spontaneous circulation. Nonsurvivors at 30 days had significantly lower mRNA levels of TLR2, IRAK3, IRAK4, NLRP3 and CASP1 in the late phase following cardiac arrest. We observed reduced proinflammatory cytokine release in response to both TLR2 and TLR4 activation in whole blood and monocyte culture assays in patients after CPR. Sera from resuscitated patients attenuated the inflammatory response in cultured PBMCs after co-stimulation with LPS. CONCLUSIONS: Successful resuscitation from cardiac arrest results in changes in monocyte pattern recognition receptor signaling pathways, which may contribute to the post-cardiac arrest syndrome. TRIAL REGISTRATION: The trial was registered in the German Clinical Trials Register ( DRKS00009684 ) on 27/11/2015.


Assuntos
Reanimação Cardiopulmonar/mortalidade , Inflamassomos/farmacocinética , Transdução de Sinais/fisiologia , Receptores Toll-Like/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/análise , Proteínas Adaptadoras de Transdução de Sinal/sangue , Idoso , Proteínas Reguladoras de Apoptose/análise , Proteínas Reguladoras de Apoptose/sangue , Proteínas Adaptadoras de Sinalização CARD , Proteínas do Citoesqueleto/análise , Proteínas do Citoesqueleto/sangue , Proteínas de Ligação a DNA/análise , Proteínas de Ligação a DNA/sangue , Feminino , Alemanha , Proteínas de Homeodomínio/análise , Proteínas de Homeodomínio/sangue , Humanos , Quinases Associadas a Receptores de Interleucina-1/análise , Quinases Associadas a Receptores de Interleucina-1/sangue , Interleucina-1beta/análise , Interleucina-1beta/sangue , Masculino , Pessoa de Meia-Idade , Monócitos/química , Monócitos/metabolismo , Monócitos/patologia , Proteína 3 que Contém Domínio de Pirina da Família NLR/análise , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Proteínas Nucleares/análise , Proteínas Nucleares/sangue , Estudos Prospectivos , Traumatismo por Reperfusão/complicações , Traumatismo por Reperfusão/etiologia , Proteínas Repressoras/análise , Proteínas Repressoras/sangue , Síndrome de Resposta Inflamatória Sistêmica/prevenção & controle , Receptor 2 Toll-Like/análise , Receptor 2 Toll-Like/sangue , Receptor 4 Toll-Like/análise , Receptor 4 Toll-Like/sangue , Fatores de Transcrição
20.
Brain Behav Immun ; 57: 125-133, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26979869

RESUMO

BACKGROUND: Inflammasomes are multimeric protein platforms involved in the regulation of inflammatory responses whose activity results in the production of proinflammatory cytokines. Because neuroinflammation is observed in autistic spectrum disorders (ASD), a neurologic condition of childhood resulting in a complex behavioural impairment, we analyzed the inflammasomes activity in ASD. Additionally we verified whether alterations of the gastrointestinal (GI) barriers might play a role in inflammasomes activation. METHODS: The activity of the inflammasomes, the concentration of the inflammasomes-derived proinflammatory cytokines interleukin (IL)-1ß and IL-18, and serum parameters of GI damage were analyzed in 25 ASD children, 23 healthy siblings (HS) and 30 unrelated age-matched healthy controls (HC). RESULTS: A significant upregulation of the AIM2 and the NLRP3 inflammasomes and an increased production of IL-1ß and IL-18 that was associated with a consistent reduction of IL-33, an anti inflammation cytokine were observed in ASD alone. Notably, in a possible immune-mediated attempt to dampen inflammation, IL-37, a suppressor of innate inflammatory responses, was significantly augmented in these same children. Finally, intestinal fatty acid binding protein (IFABP), an index of altered GI permeability, was significantly increased in serum of ASD and HS. CONCLUSIONS: These results show that the inflammasomes are activated in ASD and shed light on the molecular mechanisms responsible for ASD-associated neuroinflammation. The observation that GI alterations could be present as well in ASD offers a possible link between such alterations and neuroinflammation. Therapeutic strategies targeting inflammasome activation could be useful in ASD.


Assuntos
Transtorno do Espectro Autista/sangue , Proteínas de Ligação a DNA/sangue , Proteínas de Ligação a Ácido Graxo/sangue , Gastroenteropatias/sangue , Inflamassomos/sangue , Inflamação/sangue , Interleucinas/sangue , Proteína 3 que Contém Domínio de Pirina da Família NLR/sangue , Criança , Pré-Escolar , Feminino , Humanos , Masculino
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