RESUMO
Tumor immune microenvironment exerts a profound effect on the population of infiltrating immune cells. Tissue inhibitor of matrix metalloproteinase 1 (TIMP1) is frequently overexpressed in a variety of cells, particularly during inflammation and tissue injury. However, its function in cancer and immunity remains enigmatic. In this study, we find that TIMP1 is substantially up-regulated during tumorigenesis through analyzing cancer bioinformatics databases, which is further confirmed by IHC tissue microarrays of clinical samples. The TIMP1 level is significantly increased in lymphocytes infiltrating the tumors and correlated with cancer progression, particularly in GBM. Notably, we find that the transcriptional factor Sp1 binds to the promoter of TIMP1 and triggers its expression in GBM. Together, our findings suggest that the Sp1-TIMP1 axis can be a potent biomarker for evaluating immune cell infiltration at the tumor sites and therefore, the malignant progression of GBM.
Assuntos
Glioblastoma , Linfócitos do Interstício Tumoral , Fator de Transcrição Sp1 , Inibidor Tecidual de Metaloproteinase-1 , Carcinogênese , Linhagem Celular Tumoral , Glioblastoma/imunologia , Glioblastoma/patologia , Humanos , Linfócitos do Interstício Tumoral/imunologia , Fator de Transcrição Sp1/genética , Fator de Transcrição Sp1/imunologia , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologia , Microambiente Tumoral/imunologiaRESUMO
OBJECTIVE: To investigate the therapeutic efficacy of Tiaobu Feishen formulae (TBFS) on cigarette smoke-induced inflammation in vitro using lipopolysaccharide (LPS)-induced and cigarette smoke extract (CSE)-induced NCI-H292 cells. METHODS: We evaluated the inhibitory effects of Bufei Jianpi formula (BJF), Bufei Yishen formula (BYF), and Yiqi Zishen formula (YZF) on the expressions of inflammatory cytokines including tumor necrosis factor (TNF)-α and interleukin (IL)-8, matrix metalloproteinase (MMP)-9, tissue inhibitor of matrix metalloprotease (TIMP)-1, and superoxide dismutase (SOD) in H292 cells stimulated with LPS or CSE. Their related transcription factors and signaling pathways were also analyzed. RESULTS: BJF, BYF, and YZF significantly inhibited the LPS- or CSE-induced expressions of TNF-α, IL-8, MMP-9, TIMP-1, and SOD in H292 cells, and suppressed the activation of transcription factors including nuclear transcription factor (NF)-κB, activator protein (AP)-1, and signal transducers and activators of transcription (STAT) 3 and their corresponding pathways, including NF-κB, mitogen-activated protein kinase (MAPK), STAT3, and peroxisome proliferator-activated receptor (PPAR). CONCLUSION: BJF, BYF, and YZF effectively suppressed inflammatory responses, protease-antiprotease imbalance, and oxidative stress induced by LPS and CSE, an effect that was closely associated with the inhibition of the NF-κB, MAPK, STAT3, and PPAR pathways.
Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Células Epiteliais/efeitos dos fármacos , Pulmão/imunologia , Fumar/tratamento farmacológico , Composição de Medicamentos , Medicamentos de Ervas Chinesas/química , Células Epiteliais/metabolismo , Humanos , Interleucina-8/genética , Interleucina-8/imunologia , Pulmão/efeitos dos fármacos , NF-kappa B/genética , NF-kappa B/imunologia , Fumaça/efeitos adversos , Fumar/efeitos adversos , Fumar/genética , Fumar/imunologia , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologia , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Bleomycin-induced lung injury and fibrosis is a well-described model to investigate lung inflammatory and remodeling mechanisms. Rat models are clinically relevant and are also widely used, but rat bronchoalveolar lavage (BAL) cells are not fully characterized with flow cytometry due to the limited availability of antibodies for this species. We optimized a comprehensive time-dependent flow cytometric analysis of cells after bleomycin challenge, confirming previous studies in other species and correlating them to histological staining, cytokine profiling, and collagen accumulation analysis in rat lungs. For this purpose, we describe a novel panel of rat surface markers and a strategy to identify and follow BAL cells over time. By combining surface markers in rat alveolar cells (CD45+), granulocytes and other myeloid cells, monocytes and macrophages can be identified by the expression of CD11b/c. Moreover, different activation states of macrophages (CD163+) can be observed: steady state (CD86-MHC-IIlow), activation during inflammation (CD86+,MHC-IIhigh), activation during remodeling (CD86+MHC-IIlow), and a population of newly recruited monocytes (CD163-α-granulocyte-). Hydroxyproline measured as marker of collagen content in lung tissue showed positive correlation with the reparative phase (CD163- cells and tissue inhibitor of metalloproteinases (TIMP) and IL-10 increase). In conclusion, after a very early granulocytic recruitment, inflammation in rat lungs is observed by activated macrophages, and high release of IL-6 and fibrotic remodeling is characterized by recovery of the macrophage population together with TIMP, IL-10, and IL-18 production. Recruited monocytes and a second peak of granulocytes appear in the transitioning phase, correlating with immunostaining of arginase-1 in the tissue, revealing the importance of events leading the changes from injury to aberrant repair.
Assuntos
Lesão Pulmonar Aguda/patologia , Granulócitos/patologia , Leucócitos Mononucleares/patologia , Pulmão/patologia , Macrófagos/patologia , Monócitos/patologia , Lesão Pulmonar Aguda/induzido quimicamente , Lesão Pulmonar Aguda/genética , Lesão Pulmonar Aguda/imunologia , Animais , Antígenos CD/genética , Antígenos CD/imunologia , Arginase/genética , Arginase/imunologia , Biomarcadores/metabolismo , Bleomicina/administração & dosagem , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/imunologia , Colágeno/genética , Colágeno/imunologia , Citometria de Fluxo , Expressão Gênica , Granulócitos/efeitos dos fármacos , Granulócitos/imunologia , Interleucina-10/genética , Interleucina-10/imunologia , Interleucina-18/genética , Interleucina-18/imunologia , Interleucina-6/genética , Interleucina-6/imunologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Masculino , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Cultura Primária de Células , Ratos , Ratos Endogâmicos F344 , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologiaRESUMO
Inflammasomes are protein complexes that produce IL-1ß in response to damage or pathogens. As such, inflammasomes are involved in several types of hepatic fibrosis. However, the mechanisms by which these complexes drive the liver's fibrogenic status remain unclear. We co-cultured differentiated macrophages (the THP-1 cell line or human monocyte-derived macrophages (MDMs)) with human hepatic fibroblasts (either the LX-2 cell line or primary human hepatic stellate cells (HSCs)). The inflammasome pathway was activated with lipopolysaccharide (LPS) and monosodium urate (MSU) crystals, and the HSCs' responses were analyzed. Our results show that co-culture of HSCs with THP-1 cells upregulated transcription of the genes coding for metalloproteinase (MMP)-3 and MMP-9. After inflammasome pathway activation, the HSCs' phenotype was the same in the presence of THP-1 cells or MDMs (i.e. upregulation of MMP-3, MMP-9, and the pro-inflammatory cytokine IL-1ß). We found that two cytokines were involved in these changes: IL-1ß regulated MMP-3 and IL-1ß mRNA expression, whereas TNF-α regulated MMP-9 mRNA expression. Experiments with primary cells revealed that a general inflammatory environment is responsible for the downregulation of pro-fibrotic markers. Our present results suggest that inflammasome pathway activation in macrophages leads to a pro-inflammatory environment for HSCs leading to MMP/TIMP imbalance and enhanced fibrolytic properties.
Assuntos
Células Estreladas do Fígado/imunologia , Inflamassomos/imunologia , Macrófagos/imunologia , Metaloproteinases da Matriz/imunologia , Inibidor Tecidual de Metaloproteinase-1/imunologia , Actinas , Células Cultivadas , Humanos , Interleucina-1beta/imunologia , Fator de Necrose Tumoral alfa/imunologiaRESUMO
OBJECTIVE: Electronegative LDL (LDL(-)) is involved in atherosclerosis through the activation of the TLR4/CD14 inflammatory pathway in monocytes. Matrix metalloproteinases (MMP) and their inhibitors (tissue inhibitors of metalloproteinase [TIMP]) are also crucially involved in atherosclerosis, but their modulation by LDL(-) has never been investigated. The aim of this study was to examine the ability of LDL(-) to release MMPs and TIMPs in human monocytes and to determine whether sulodexide (SDX), a glycosaminoglycan-based drug, was able to affect their secretion. APPROACH AND RESULTS: Native LDL (LDL(+)) and LDL(-) separated by anion-exchange chromatography were added to THP1-CD14 monocytes in the presence or absence of SDX for 24â¯h. A panel of 9 MMPs and 4 TIMPs was analyzed in cell supernatants with multiplex immunoassays. The gelatinolytic activity of MMP-9 was assessed by gelatin zymography. LDL(-) stimulated the release of MMP-9 (13-fold) and TIMP-1 (4-fold) in THP1-CD14 monocytes, as well as the gelatinolytic activity of MMP-9. Co-incubation of monocytes with LDL(-) and SDX for 24â¯h significantly reduced both the release of MMP-9 and TIMP-1 and gelatinase activity. In THP1 cells not expressing CD14, no effect of LDL(-) on MMP-9 or TIMP-1 release was observed. The uptake of DiI-labeled LDL(-) was higher than that of DiI-LDL(+) in THP1-CD14 but not in THP1 cells. This increase was inhibited by SDX. Experiments in microtiter wells coated with SDX demonstrated a specific interaction of LDL(-) with SDX. CONCLUSIONS: LDL(-) induced the release of MMP-9 and TIMP-1 in monocytes through CD14. SDX affects the ability of LDL(-) to promote TIMP-1 and MMP-9 release by its interaction with LDL(-).
Assuntos
Anti-Inflamatórios/farmacologia , Glicosaminoglicanos/farmacologia , Receptores de Lipopolissacarídeos/imunologia , Lipoproteínas LDL/imunologia , Metaloproteinase 9 da Matriz/imunologia , Monócitos/efeitos dos fármacos , Inibidor Tecidual de Metaloproteinase-1/imunologia , Linhagem Celular , Humanos , Hipolipemiantes/farmacologia , Lipoproteínas LDL/química , Monócitos/imunologia , Eletricidade EstáticaRESUMO
Mycotoxin T-2 exerts a causative role in Kashin-Beck disease (KBD) suffering chondrocyte apoptosis and cartilage matrix homeostasis disruption. Recent research corroborated the aberrant levels of pro-inflammatory cytokine IL-1ß in KBD patients and mycotoxin environment. In the present study, we investigated the relevance of IL-1ß in T-2 toxin-evoked chondrocyte cytotoxic injury and aberrant catabolism. High levels of IL-1ß were detected in serum and cartilages from KBD patients and in T-2-stimulated chondrocytes. Moreover, knockdown of IL-1ß antagonized the adverse effects of T-2 on cytotoxic injury by enhancing cell viability and inhibiting apoptosis. However, exogenous supplementation of IL-1ß further aggravated cell damage in response to T-2. Additionally, cessation of IL-1ß rescued T-2-elicited tilt of matrix homeostasis toward catabolism by elevating the transcription of collagen II and aggrecan, promoting release of sulphated glycosaminoglycans (sGAG) and TIMP1, and suppressing matrix metalloproteinases production including MMP-1, MMP-3 and MMP-13. Conversely, IL-1ß stimulation deteriorated T-2-induced disruption of matrix metabolism balance toward catabolism. Mechanistic analysis found the high activation of Wnt/ß-catenin in KBD patients and chondrocytes upon T-2. Furthermore, this activation was mitigated after IL-1ß inhibition, but further enhanced following IL-1ß precondition. Importantly, blocking this pathway by transfection with ß-catenin alleviated the adverse roles of IL-1ß on cytotoxic injury and metabolism disorders under T-2 conditioning. Together, this study elucidates a new insight into how T-2 deteriorates the pathological progression of KBD by regulating inflammation-related pathways, indicating a promising anti-inflammation strategy for KBD therapy.
Assuntos
Condrócitos/imunologia , Interleucina-1beta/imunologia , Toxina T-2/toxicidade , Via de Sinalização Wnt/efeitos dos fármacos , beta Catenina/imunologia , Adulto , Agrecanas/biossíntese , Agrecanas/genética , Agrecanas/imunologia , Animais , Apoptose/genética , Apoptose/imunologia , Condrócitos/metabolismo , Condrócitos/patologia , Colágeno Tipo II/biossíntese , Colágeno Tipo II/genética , Colágeno Tipo II/imunologia , Colagenases/biossíntese , Colagenases/genética , Colagenases/imunologia , Matriz Extracelular/genética , Matriz Extracelular/imunologia , Matriz Extracelular/metabolismo , Matriz Extracelular/patologia , Feminino , Humanos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Doença de Kashin-Bek/genética , Doença de Kashin-Bek/imunologia , Doença de Kashin-Bek/metabolismo , Doença de Kashin-Bek/patologia , Masculino , Pessoa de Meia-Idade , Ratos , Ratos Sprague-Dawley , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologia , Transcrição Gênica/efeitos dos fármacos , Transcrição Gênica/imunologia , Via de Sinalização Wnt/genética , Via de Sinalização Wnt/imunologia , beta Catenina/genética , beta Catenina/metabolismoRESUMO
OBJECTIVES: Zinc (Zn) has major effects on immune system activation while Cadmium (Cd) has anti-inflammatory and anti-proliferative effects in several chronic inflammatory contexts. The aim of this work was to investigate by which mechanisms Zn could compete with Cd and eventually counteract its deleterious effects. Rheumatoid arthritis (RA) synoviocytes exposed to cytokines were used as a model of chronic inflammation; osteoarthritis (OA) synoviocytes were used as control. METHODS: Cell/medium fractionation constants were analyzed for different metals by inductively-coupled-plasma mass-spectrometry by comparison to the 70Zn spike. Interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-α) were used to mimic inflammation. Gene expression of ZIP-8 importer, metallothioneins-1 (MT-1s) and the ratio between metalloprotease-3 and the tissue inhibitor of metalloproteinases (MMP-3)/TIMP-1) were evaluated after pre-exposure to cytokines and Cd, with or without the addition of exogenous Zn (0.9 ppm). Cell viability was measured by neutral red assay and IL-6 production by ELISA. RESULTS: Synoviocytes selectively absorbed and retained Cd in comparison to Zn. Metal import increased with IL-17/TNF-α exposure, through the enhanced ZIP-8 expression. Zn did not modify ZIP-8 expression, while Cd reduced it (p<0.05). Zn induced a reduction of Cd-induced MT-1s expression, in particular of MT-1X (3-fold), and subsequently the final intra-cellular content of Cd. By reducing Cd accumulation in cells, Zn reversed Cd anti-proliferative and anti-inflammatory effects but preserved the low MMP-3/TIMP-1 ratio induced by Cd, which was enhanced by inflammatory conditions. CONCLUSION: Zinc counteracts the deleterious effect of Cd by reducing its import and accumulation in the cell, without the reactivation of destructive pathways such as MMPs.
Assuntos
Artrite Reumatoide/imunologia , Cádmio/imunologia , Osteoartrite/imunologia , Zinco/imunologia , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Transporte Biológico , Cádmio/metabolismo , Proliferação de Células , Células Cultivadas , Doença Crônica , Humanos , Interleucina-17/imunologia , Metaloproteinase 3 da Matriz/imunologia , Metalotioneína/imunologia , Osteoartrite/metabolismo , Osteoartrite/patologia , Membrana Sinovial/citologia , Membrana Sinovial/imunologia , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia , Inibidor Tecidual de Metaloproteinase-1/imunologia , Fator de Necrose Tumoral alfa/imunologia , Zinco/metabolismoRESUMO
OBJECTIVES: Multiple sclerosis (MS) is an autoimmune disease involving the central nervous system (CNS) with unknown immunopathogenic mechanisms. Matrix metalloproteinase-9 (MMP-9) facilitates T-cell migration into the CNS while the tissue inhibitor matrix metalloproteinase-1 (TIMP-1) inhibits MMP-9 actions. The aim of this study was to evaluate the expression of TIMP-1 RNA and MMP-9/TIMP-1 RNA ratio in blood cells of Iranian patients with relapsing-remitting multiple sclerosis (RRMS) treated with IFNb. MATERIAL AND METHODS: The study compared the expression level of TIMP-1 gene in RRMS samples with normal individuals in Iran and the results were compared using a ratio of MMP-9 to TIMP-1. All patients were HLA-DRB1*15 negative and were responders to interferon-beta with a normal vitamin D level. RESULTS: The RRMS patients manifested a lower expression level of TIMP-1 RNA than their normal counterparts although the result was not significant (P= 0.06). Also, the ratio of MMP-9 to TIMP-1 RNA increased significantly (P= 0.009). There was no linear correlation between TIMP-1 expression level and risk of Expanded Disability Status Scale of Kurtzke (EDSS); nor was there any significant correlation between expression status of TIMP-1 and duration of the disease. Although there was no significant decrease in TIMP-1 expression level, the MMP-9/TIMP-1 RNA ratio in RRMS was significantly higher than normal subjects. CONCLUSION: Further studies are recommended to compare MMP-9/TIMP-1 RNA ratio in patients before and after taking IFN-beta in order to find out if MMP-9/TIMP-1 RNA ratio can function as a proper marker of the bio efficacy of IFN-beta treatment of MS.
Assuntos
Metaloproteinase 9 da Matriz/genética , Esclerose Múltipla Recidivante-Remitente/genética , RNA Mensageiro/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Adolescente , Adulto , Idoso , Estudos de Casos e Controles , Feminino , Regulação da Expressão Gênica , Cadeias HLA-DRB1/genética , Cadeias HLA-DRB1/imunologia , Humanos , Fatores Imunológicos/uso terapêutico , Interferon beta/uso terapêutico , Masculino , Metaloproteinase 9 da Matriz/imunologia , Pessoa de Meia-Idade , Esclerose Múltipla Recidivante-Remitente/tratamento farmacológico , Esclerose Múltipla Recidivante-Remitente/imunologia , Esclerose Múltipla Recidivante-Remitente/patologia , RNA Mensageiro/imunologia , Reação em Cadeia da Polimerase em Tempo Real , Inibidor Tecidual de Metaloproteinase-1/imunologia , Vitamina D/sangueRESUMO
Extracellular matrix metalloproteinase inducer (EMMPRIN, CD147) is a transmembrane glycoprotein that is upregulated on leukocytes in active lesions in multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE). Administration of anti-EMMPRIN Abs reduces the severity of EAE. Minocycline is a tetracycline antibiotic with immune-modulatory properties that decreases the severity of EAE; it was recently found to attenuate the conversion from a first demyelinating event to clinically definite MS in a phase III trial. We investigated whether and how minocycline affects the expression of EMMPRIN on T cells in culture and in mice afflicted with EAE. EMMPRIN expression in cultures of mouse splenocytes or human PBMCs was elevated upon polyclonal T cell activation, and this was reduced by minocycline correspondent with decreased P-Akt levels. An established MS medication, IFN-ß, also diminished EMMPRIN levels on human cells whereas this was not readily observed for fingolimod or monomethylfumarate. In EAE-afflicted mice, minocycline treatment significantly reduced EMMPRIN levels on splenic lymphocytes at the presymptomatic (day 7) phase, and prevented the development of disease. Day 7 spleen transcripts from minocycline-treated EAE mice had a significantly lower MMP-9/TIMP-1 ratio, and significantly lower MCT-1 and CD98 levels, factors associated with EMMPRIN function. Day 16 (peak clinical severity) CNS samples from EAE mice had prominent representation of inflammatory perivascular cuffs, inflammatory molecules and EMMPRIN, and these were abrogated by minocycline. Overall, minocycline attenuated the activation-induced elevation of EMMPRIN on T cells in culture and in EAE mice, correspondent with reduced immune function and EAE CNS pathology.
Assuntos
Antibacterianos/farmacologia , Basigina/metabolismo , Encefalomielite Autoimune Experimental/tratamento farmacológico , Encefalomielite Autoimune Experimental/imunologia , Minociclina/uso terapêutico , Esclerose Múltipla/imunologia , Linfócitos T/efeitos dos fármacos , Animais , Antibacterianos/administração & dosagem , Antibacterianos/uso terapêutico , Basigina/genética , Sistema Nervoso Central/efeitos dos fármacos , Ensaios Clínicos Fase III como Assunto , Modelos Animais de Doenças , Encefalomielite Autoimune Experimental/prevenção & controle , Cloridrato de Fingolimode/farmacologia , Fumaratos/farmacologia , Humanos , Interferon beta/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Maleatos/farmacologia , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Minociclina/administração & dosagem , Minociclina/farmacologia , Monócitos , Esclerose Múltipla/genética , Esclerose Múltipla/fisiopatologia , Linfócitos T/imunologia , Linfócitos T/metabolismo , Inibidor Tecidual de Metaloproteinase-1/imunologiaRESUMO
OBJECTIVE: The aim of this study was to longitudinally evaluate and analyze the role of interleukin-22-producing CD4 positive cells (IL-22) in the pathogenesis of Hepatitis C Virus recurrence after Orthotopic Liver Transplantation (HCV-OLT). METHODS: 15 HCV-OLT, 15 age- and gender- matched non-HCV post-OLT (OLT) and 15 hepatitis C virus infected (HCV) patients were enrolled into our study from the liver transplantation and research center at Beijing 302 Hospital. We determined the frequencies of IL-22 using flow cytometry and expression of IL-22 mRNA using PCR in peripheral blood and liver tissue. We also divided HCV-OLT patients into rapid fibrosis progression (RFP) and slow fibrosis progression (SFP), examined IL-22 cells and analyzed the correlations between IL-22 frequencies and liver injury, fibrosis and clinical parameters. Moreover, we investigated the role of IL-22 in Human Hepatic Stellate Cells (HSCs). RESULTS: The levels of serum IL-22, frequencies of IL-22 producing cells in peripheral blood mononuclear cells, and expression of IL-22 mRNA and protein in the liver in the HCV-OLT group were significantly higher than that in the HCV and OLT groups. Furthermore, eight (53.3%) patients developed RFP after two years; another three patients were diagnosed liver cirrhosis. The frequencies of IL-22 were much higher in RFP compared with SFP, while no significant difference existed between OLT and SFP. Intrahepatic IL-22 positive cells were located in fibrotic areas and significantly correlated with α-smooth muscle actin (α-SMA) and fibrosis staging scores, not with grading scores and HCRVNA. In vitro, IL-22 administration prevented HSCs apoptosis, promoted HSCs proliferation and activation, up-regulated the expression of HSC-sourced growth factors including α-SMA, TGF-ß and TIMP-1, and increased the production of liver fibrosis markers including laminin, hyaluronic acid and collagen type IV. CONCLUSION: Peripheral and intrahepatic IL-22 is up-regulated and plays a pathological role in exacerbating liver fibrosis by activating HSCs in HCV-OLT patients, which may predict RFP and serve as an attractive target for anti-fibrotic therapy.
Assuntos
Células Estreladas do Fígado/patologia , Hepatite C Crônica/patologia , Interleucinas/genética , Cirrose Hepática/patologia , Transplante de Fígado , Fígado/patologia , Actinas/genética , Actinas/imunologia , Adulto , Colágeno Tipo IV/genética , Colágeno Tipo IV/imunologia , Progressão da Doença , Feminino , Expressão Gênica , Hepacivirus/imunologia , Hepacivirus/patogenicidade , Células Estreladas do Fígado/imunologia , Células Estreladas do Fígado/virologia , Hepatite C Crônica/genética , Hepatite C Crônica/imunologia , Hepatite C Crônica/virologia , Humanos , Ácido Hialurônico/biossíntese , Ácido Hialurônico/imunologia , Interleucinas/imunologia , Laminina/genética , Laminina/imunologia , Fígado/imunologia , Fígado/cirurgia , Fígado/virologia , Cirrose Hepática/genética , Cirrose Hepática/imunologia , Cirrose Hepática/virologia , Masculino , Pessoa de Meia-Idade , RNA Mensageiro/genética , RNA Mensageiro/imunologia , Recidiva , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologia , Fator de Crescimento Transformador beta/genética , Fator de Crescimento Transformador beta/imunologia , Carga Viral/imunologia , Interleucina 22RESUMO
AIM: The aim of this work was to evaluate the effects of carnosol, a rosemary polyphenol, on pro-inflammatory and catabolic mediators of cartilage breakdown in chondrocytes and via bone-cartilage crosstalk. MATERIALS AND METHODS: Osteoarthritic (OA) human chondrocytes were cultured in alginate beads for 4 days in presence or absence of carnosol (6 nM to 9 µM). The production of aggrecan, matrix metalloproteinase (MMP)-3, tissue inhibitor of metalloproteinase (TIMP)-1, interleukin (IL)-6 and nitric oxide (NO) and the expression of type II collagen and ADAMTS-4 and -5 were analyzed. Human osteoblasts from sclerotic (SC) or non-sclerotic (NSC) subchondral bone were cultured for 3 days in presence or absence of carnosol before co-culture with chondrocytes. Chondrocyte gene expression was analyzed after 4 days of co-culture. RESULTS: In chondrocytes, type II collagen expression was significantly enhanced in the presence of 3 µM carnosol (p = 0.008). MMP-3, IL-6, NO production and ADAMTS-4 expression were down-regulated in a concentration-dependent manner by carnosol (p<0.01). TIMP-1 production was slightly increased at 3 µM (p = 0.02) and ADAMTS-5 expression was decreased from 0.2 to 9 µM carnosol (p<0.05). IL-6 and PGE2 production was reduced in the presence of carnosol in both SC and NSC osteoblasts while alkaline phosphatase activity was not changed. In co-culture experiments preincubation of NSC and SC osteoblasts wih carnosol resulted in similar effects to incubation with anti-IL-6 antibody, namely a significant increase in aggrecan and decrease in MMP-3, ADAMTS-4 and -5 gene expression by chondrocytes. CONCLUSIONS: Carnosol showed potent inhibition of pro-inflammatory and catabolic mediators of cartilage breakdown in chondrocytes. Inhibition of matrix degradation and enhancement of formation was observed in chondrocytes cocultured with subchondral osteoblasts preincubated with carnosol indicating a cross-talk between these two cellular compartments, potentially mediated via inhibition of IL-6 in osteoblasts as similar results were obtained with anti-IL-6 antibody.
Assuntos
Abietanos/farmacologia , Anti-Inflamatórios/farmacologia , Condrócitos/efeitos dos fármacos , Condrócitos/patologia , Osteoartrite/tratamento farmacológico , Osteoartrite/imunologia , Agrecanas/imunologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Condrócitos/imunologia , Técnicas de Cocultura , Dinoprostona/imunologia , Humanos , Interleucina-6/imunologia , Metaloproteinase 3 da Matriz/imunologia , Osteoartrite/patologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/imunologia , Osteoblastos/patologia , Inibidor Tecidual de Metaloproteinase-1/imunologiaRESUMO
Dengue, the most rampant zoonotic viral disease in tropics, contributes to 14% of acute febrile illness cases globally. Encephalitis in primary Dengue fever, with/without haemorrhage has been reported occasionally. Our study presents novel evidence for this rarity at the molecular level. Murine microglia (BV2) were infected in-vitro with Dengue virus (DENV) serotypes (1-4) and their immune response was evaluated. Gene expressions of TNF-α, IL-10, IFN-γ, and IL1-ß constituted the pro-inflammatory response, levels of MCP-1 and IL-6 represented the regulatory mechanism and changes in the levels of Occludin, MMP-2, MMP-9 and TIMP-1 encompassed the break-down of the blood-brain barrier (BBB). Cytokine response was studied using RT-PCR, with relative fold change assessed using ΔΔCt method. We observed that DENV1 increased vascular permeability and trans-membrane transport, while DENV2 resulted in oxidative stress. DENV3 infection presented with impaired immune response and DENV4 manifested a chaotropic response of the BBB protein genes. However, no serotype was able to breakdown the BBB, thus validating the low prevalence of encephalitis in dengue. Our study is the first reported evidence of the microglial immune response resisting the entry of DENV into the CNS. It also supports the theory that primary Dengue infection results in the acute inflammation of the microglia, and the host immune response plays a critical role in development of encephalitis.
Assuntos
Barreira Hematoencefálica/imunologia , Vírus da Dengue/imunologia , Dengue/imunologia , Microglia/imunologia , Animais , Barreira Hematoencefálica/patologia , Barreira Hematoencefálica/virologia , Linhagem Celular , Citocinas/imunologia , Dengue/patologia , Metaloproteinase 2 da Matriz/imunologia , Metaloproteinase 9 da Matriz/imunologia , Camundongos , Microglia/patologia , Microglia/virologia , Inibidor Tecidual de Metaloproteinase-1/imunologiaRESUMO
BACKGROUND: Inflammatory dilated cardiomyopathy (iDCM) is a common debilitating disease with poor prognosis that often leads to heart failure and may require heart transplantation. The aim of this study was to evaluate sera and biopsy samples from chronic iDCM patients, and to investigate molecular mechanism associated with left ventricular remodeling and disease progression in order to improve therapeutic intervention. METHODS: Patients were divided into inflammatory and non-inflammatory DCM groups according to the immunohistochemical expression of inflammatory infiltrates markers: T-lymphocytes (CD3), active-memory T lymphocyte (CD45Ro) and macrophages (CD68). The inflammation, apoptosis, necrosis and fibrosis were investigated by ELISA, chemiluminescent, immunohistochemical and histological assays. RESULTS: The pro-inflammatory cytokine IL-6 was significantly elevated in iDCM sera (3.3 vs. 10.98 µg/ml; P < 0.05). Sera levels of caspase-9, -8 and -3 had increased 6.24-, 3.1- and 3.62-fold, (P < 0.05) and only slightly (1.3-, 1.22- and 1.03-fold) in biopsies. Significant release of Hsp60 in sera (0.0419 vs. 0.36 ng/mg protein; P < 0.05) suggested a mechanistic involvement of mitochondria in cardiomyocyte apoptosis. The significant MMP9/TIMP1 upregulation in biopsies (0.1931 - 0.476, P < 0.05) and correlation with apoptosis markers show its involvement in initiation of cell death and ECM degradation. A slight activation of the extrinsic apoptotic pathway and the release of hsTnT might support the progression of chronic iDCM. CONCLUSIONS: Data of this study show that significant increase of IL-6, MMP9/TIMP1 and caspases-9, -8, -3 in sera corresponds to molecular mechanisms dominating in chronic iDCM myocardium. The initial apoptotic pathway was more activated by the intramyocardial inflammation and might be associated with extrinsic apoptotic pathway through the pro-apoptotic Bax. The activated intrinsic form of myocardial apoptosis, absence of necrosis and decreased fibrosis are most typical characteristics of chronic iDCM. Clinical use of anti-inflammatory drugs together with specific anti-apoptotic treatment might improve the efficiency of therapies against chronic iDCM before heart failure occurs.
Assuntos
Apoptose/imunologia , Cardiomiopatia Dilatada/imunologia , Fibrose/imunologia , Inflamação/imunologia , Macrófagos/imunologia , Miocárdio/imunologia , Necrose/imunologia , Linfócitos T/imunologia , Remodelação Ventricular/imunologia , Adulto , Antígenos CD/imunologia , Antígenos de Diferenciação Mielomonocítica/imunologia , Complexo CD3/imunologia , Cardiomiopatia Dilatada/metabolismo , Cardiomiopatia Dilatada/patologia , Caspase 3/imunologia , Caspase 8/imunologia , Caspase 9/imunologia , Chaperonina 60/imunologia , Citocinas/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imuno-Histoquímica , Inflamação/patologia , Antígenos Comuns de Leucócito/imunologia , Masculino , Metaloproteinase 9 da Matriz/imunologia , Pessoa de Meia-Idade , Proteínas Mitocondriais/imunologia , Miocárdio/metabolismo , Miocárdio/patologia , Subpopulações de Linfócitos T/imunologia , Inibidor Tecidual de Metaloproteinase-1/imunologia , Troponina T/metabolismoRESUMO
Macrophages are important in vascular inflammation and environmental factors influence macrophage plasticity. Macrophage transitions into pro-inflammatory (M1) or anti-inflammatory (M2) states have been defined predominately by measuring cytokines in culture media (CM). However, temporal relationships between cellular and secreted cytokines have not been established. We measured phenotypic markers and cytokines in cellular and CM of murine bone marrow-derived macrophages at multiple time points following stimulation with IFN-γ + LPS (M1), IL-4 (M2a) or IL-10 (M2c). Cytokines/proteins in M1-polarized macrophages exhibited two distinct temporal patterns; an early (0.5-3 h), transient increase in cellular cytokines (GM-CSF, KC-GRO, MIP-2, IP-10 and MIP-1ß) and a delayed (3-6 h) response that was more sustained [IL-3, regulated on activation normal T cell expressed and secreted (RANTES), and tissue inhibitor of metalloproteinases 1 (TIMP-1)]. M2a-related cytokine/cell markers (IGF-1, Fizz1 and Ym1) were progressively (3-24 h) increased post-stimulation. In addition, novel patterns were observed. First, and unexpectedly, cellular pro-inflammatory chemokines, MCP-1 and MCP-3 but not MCP-5, were comparably increased in M1 and M2a macrophages. Second, Vegfr1 mRNA was decreased in M1 and increased in M2a macrophages. Finally, VEGF-A was increased in the CM of M1 cultures and strikingly reduced in M2a coinciding with increased Vegfr1 expression, suggesting decreased VEGF-A in M2a CM was secondary to increased soluble VEGFR1. In conclusion, macrophage cytokine production and marker expression were temporally regulated and relative levels compared across polarizing conditions were highly dependent upon the timing and location (cellular versus CM) of the sample collection. For most cytokines, cellular production preceded increases in the CM suggesting that cellular regulatory pathways should be studied within 6 h of stimulation. The divergent polarization-dependent expression of Vegfr1 may be essential to controlling VEGF potentially regulating angiogenesis and inflammatory cell infiltration in the vascular niche. The current study expands the repertoire of cytokines produced by polarized macrophages and provides insights into the dynamic regulation of macrophage polarization and resulting cytokines, proteins and gene expression that influence vascular inflammation.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Macrófagos/efeitos dos fármacos , Fenótipo , Fator A de Crescimento do Endotélio Vascular/imunologia , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/imunologia , Animais , Quimiocina CCL2/genética , Quimiocina CCL2/imunologia , Quimiocina CCL4/genética , Quimiocina CCL4/imunologia , Quimiocina CCL7/genética , Quimiocina CCL7/imunologia , Quimiocina CXCL10/genética , Quimiocina CXCL10/imunologia , Quimiocina CXCL2/genética , Quimiocina CXCL2/imunologia , Regulação da Expressão Gênica , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Humanos , Interferon gama/farmacologia , Interleucina-10/farmacologia , Interleucina-4/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/citologia , Macrófagos/imunologia , Camundongos , Cultura Primária de Células , Transdução de Sinais , Fatores de Tempo , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologia , Fator A de Crescimento do Endotélio Vascular/genética , Receptor 1 de Fatores de Crescimento do Endotélio Vascular/genéticaRESUMO
We investigated the inhibitory effects of Platycodon grandiflorum root-derived saponins (Changkil saponins: CKS) on ovalbumin-induced airway inflammation in mice. CKS suppressed leukocytes number, IgE, Th1/Th2 cytokines, and MCP-1 chemokine secretion in bronchoalveolar lavage fluid. Also, ovalbumin-increased MUC5AC, MMP-2/9, and TIMP-1/-2 mRNA expression, NF-κB activation, leukocytes recruitment, and mucus secretion were inhibited by CKS treatment. Moreover, the active component of CKS, platyconic acid A (PA), suppressed PMA-induced MUC5AC mRNA expression (from 2.1 ± 0.2 to 1.1 ± 0.1) by inhibiting NF-κB activation (from 2.3 ± 0.2 to 1.2 ± 0.1) via Akt (from 3.7 ± 0.3 to 2.1 ± 0.2) (p < 0.01) in A549 cells. Therefore, we demonstrate that CKS or PA suppressed the development of respiratory inflammation, hyperresponsiveness, and remodeling by reducing allergic responses, and they may be potential herbal drugs for allergen-induced respiratory disease prevention.
Assuntos
Anti-Inflamatórios/administração & dosagem , Pulmão/imunologia , Platycodon/química , Saponinas/administração & dosagem , Triterpenos/administração & dosagem , Animais , Linhagem Celular , Citocinas/genética , Citocinas/imunologia , Feminino , Humanos , Pulmão/efeitos dos fármacos , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/imunologia , Camundongos , Camundongos Endogâmicos ICR , Mucina-5AC/genética , Mucina-5AC/imunologia , Ovalbumina/efeitos adversos , Raízes de Plantas/química , Acetato de Tetradecanoilforbol/efeitos adversos , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/imunologiaRESUMO
UNLABELLED: The aim of study was to determine the leading clinical, immunological and sonographic pararneters, reflecting the efficiency of Ursodeoxycholic acid (UDCA) at the rate of 10 mg per 1 kg of body weight in the treatment of gallstone disease in patients with metabolic syndrome (MS). MATERIALS AND METHODS: An assessment of clinical, biochemical immunological and sonographic parameters in 54 patients with gallstone disease associated with the metabolic syndrome before and after the six-month treatment UDCA were made. RESULTS: In accordance with our results the significant predictors, reflecting successful litholitic therapy at patients with gallstone disease in association with metabolic syndrome are decrease the serum concentration of gamma-glutamyltranspeptidase (P = 0.003), matrix metalloproteinase-9 (P = 0.001), increase the serum concentration of tissue inhibitor of metalloproteinases-1 (P = 0.02), decrease the left liver lobe thickness (P = 0,003) and the thickness of gallbladder wall (P = 0.0002). CONCLUSION: The results of our study have shown that the therapy with ursodesoxycholic acid of patients with metabolic syndrome leads to decrease of factors of gallstone progression (elevated levels of gamma-glutamyltranspeptidase, matrix metalloproteinase-9 and increased thickness of the left lobe liver and gallbladder wall).
Assuntos
Cálculos Biliares , Metaloproteinase 9 da Matriz , Síndrome Metabólica , Inibidor Tecidual de Metaloproteinase-1 , Ácido Ursodesoxicólico/administração & dosagem , gama-Glutamiltransferase , Adulto , Idoso , Feminino , Cálculos Biliares/sangue , Cálculos Biliares/complicações , Cálculos Biliares/diagnóstico por imagem , Cálculos Biliares/tratamento farmacológico , Cálculos Biliares/imunologia , Humanos , Masculino , Metaloproteinase 9 da Matriz/sangue , Metaloproteinase 9 da Matriz/imunologia , Síndrome Metabólica/sangue , Síndrome Metabólica/complicações , Síndrome Metabólica/diagnóstico por imagem , Síndrome Metabólica/tratamento farmacológico , Síndrome Metabólica/imunologia , Pessoa de Meia-Idade , Inibidor Tecidual de Metaloproteinase-1/sangue , Inibidor Tecidual de Metaloproteinase-1/imunologia , Ultrassonografia , gama-Glutamiltransferase/sangue , gama-Glutamiltransferase/imunologiaRESUMO
Os ameloblastomas e tumores odontogênicos ceratocísticos (TOC) representam lesões odontogênicas que, apesar de sua natureza benigna, se destacam por um comportamento biológico distinto, caracterizado pelo crescimento localmente agressivo e episódios recidivantes. A reabsorção dos ossos gnáticos provocada pelo crescimento dessas lesões constitui um fator determinante à expansão das mesmas, sendo mediada tanto por células osteoclásticas como pela ação enzimática de diversas metaloproteinases de matriz (MMPs). A expressão de fatores estimuladores e inibidores da reabsorção óssea vem sendo correlacionada com o desenvolvimento destas lesões, merecendo destaque algumas MMPs como as colagenases e as gelatinases e os inibidores teciduais de metaloproteinases (TIMPs), dentre outros. Baseados na premissa de que fatores estimuladores e inibidores de processos osteolíticos podem ser determinantes para o ritmo de crescimento de lesões odontogênicas intraósseas, o objetivo de estudo foi avaliar a imunoexpressão das proteínas MMP-9, -13 e TIMP-1 no epitélio e mesênquima de espécimes de ameloblastomas e TOC. A análise estatística foi realizada através dos testes de Mann-Whitney e Wilcoxon com nível de significância estabelecido em 5%. Através de uma análise quantitativa das células imunomarcadas, foi observada a expressão imuno-histoquímica das MMP-9, -13 e TIMP-1 em 100% dos casos, tanto no epitélio quanto no mesênquima tumoral. Mais de 76% das células epiteliais (escore 3) dos TOC e ameloblastomas apresentaram imunomarcação para MMP-9 (p=0,382) e MMP-13 (p=0,069), sendo estatisticamente significativa para o TIMP-1 (p=0,003) nos ameloblastomas. No mesênquima, observou-se maior escore de imunomarcação da MMP-13 (p=0,031) nos ameloblastomas em relação aos TOC, enquanto para a MMP-9 e TIMP-1 não se observou diferença estatisticamente significativa (p=0,403; p=1,000). O cálculo da razão entre os escores de expressão das proteínas revelou, de uma maneira geral, similaridade entre as lesões, sendo observado predomínio significante de igualdade de expressão do TIMP-1 e da MMP-9 apenas no epitélio dos ameloblastomas. A imunoexpressão marcante das MMP-9, MMP-13 e TIMP-1 no epitélio e mesênquima das lesões estudadas indica que estas proteínas participam na remodelação da MEC necessária à progressão tumoral, no entanto, as diferenças pontuais observadas na expressão de algumas destas proteínas, não são suficientes para sugerir diferenças no comportamento biológico dos ameloblastomas e dos TOCs. (AU)
Ameloblastomas and keratocystic odontogenic tumors (KOT) represent odontogenic lesions that, despite their benign nature, are distinguished by a distinct biological behavior, characterized by locally aggressive growth and recurrent episodes. The gnathic bone resorption caused by the growth of these lesions is a key to the expansion of the same, both being mediated by osteoclastic cells like enzymatic activity of various matrix metalloproteinases (MMPs) factor. The expression of stimulatory factors and inhibitors of bone resorption has been correlated with the development of these lesions, with emphasis to some MMPs such as collagenases and gelatinases and tissue inhibitors of metalloproteinases (TIMPs), among others. Based on the premise that stimulatory and inhibitory factors of osteolytic processes can be decisive for the growth rate of intraosseous odontogenic lesions, this experiment evaluated the immunoreactivity of MMP-9, -13 and TIMP-1 protein in the epithelium and mesenchyme of ameloblastoma and the KOT specimens, by a quantitative analysis of the immunoreactivity cells. Statistical analysis was performed using the MannWhitney and Wilcoxon tests with a significance level set at 5 %. Immunohistochemical expression of MMP-9, -13 and TIMP-1 was observed in 100% of cases both in the epithelium and in mesenchyme. The immunoreactivity in the epithelium of KOT and ameloblastomas revealed a predominance of score 3 for MMP-9 (p=0.382) and MMP-13 (p=0.069) and no statistically significance for TIMP-1, the latter being significantly higher immunoreactivity in ameloblastomas. In the mesenchyme, there was a higher score immunoreactivity of MMP-13 (p=0.031) in ameloblastomas in relation to KOT, whereas for MMP-9 and TIMP-1 no statistically significant difference (p=0.403 was observed, p=1.000). The calculation of the ratio of scores revealed expression of proteins in general, similarity of the lesions, a significant predominance of equal expression of TIMP-1 and MMP-9 was observed only in the epithelium of ameloblastoma. The marked immunostaining of MMP-9 , MMP-13 and TIMP-1 in epithelium and mesenchyme of the lesion indicate that these proteins involved in ECM remodeling required for tumor progression, however, specific differences in the expression of some of these proteins, are not sufficient to suggest differences in the biological behavior of ameloblastomas and KOTs. (AU)
Assuntos
Ameloblastoma/imunologia , Cistos Odontogênicos/imunologia , Inibidor Tecidual de Metaloproteinase-1/imunologia , /imunologia , Metaloproteinase 9 da Matriz/imunologia , Estatísticas não Paramétricas , Imuno-Histoquímica/métodos , Microscopia/métodos , Reabsorção ÓsseaRESUMO
BACKGROUND: Tissue transglutaminase (tTG) is a post-translational modifying enzyme located in airway epithelial cells. A potential contribution of serum specific IgG (sIgG) to tTG in airway inflammation of toluene diisocyanate (TDI)-induced occupational asthma (OA) has been suggested. OBJECTIVE: To prepare a TDI-tTG conjugate and detect serum specific antibodies in sera of patients with TDI-OA to understand this mechanism. METHODS: Ninety-nine patients with TDI-OA, 76 asymptomatic exposed controls, 208 patients with non-OA, and 74 unexposed controls were enrolled for this study. The TDI-tTG conjugate was prepared and confirmed by a native gel. Serum sIgG and/or sIgE antibodies to tTG, TDI-tTG, TDI conjugated to human serum albumin, cytokeratin 19, and serum cytokine levels, such as interleukin-8, transforming growth factor-ß1, and tissue inhibitor of metalloproteinase-1, were measured by enzyme-linked immunosorbent assay. The level of interleukin-8 produced from airway epithelial cells (A549) treated with tTG was evaluated to investigate the inflammatory effect of tTG and TDI-tTG. RESULTS: In the TDI-OA group, the prevalence of serum sIgG to TDI-tTG (17.2%) was higher than that of sIgG to tTG (11.1%), which were significantly higher than those of the 3 control groups (P < .05 for all groups). TDI-exposed subjects with high levels of serum sIgG to TDI-tTG had a high prevalence of sIgG to cytokeratin 19 and higher serum levels of transforming growth factor-ß1 and tissue inhibitor of metalloproteinase-1. The tTG and TDI-tTG dose-dependently increased interleukin-8 production from A549 cells. CONCLUSION: These findings suggest that TDI exposure in the workplace binds to tTG to form a conjugate that can induce serum sIgG antibody production, airway inflammation, and airway remodeling in patients with TDI-OA.
Assuntos
Remodelação das Vias Aéreas/efeitos dos fármacos , Asma Ocupacional/sangue , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Tolueno 2,4-Di-Isocianato/efeitos adversos , Transglutaminases/efeitos adversos , Adulto , Remodelação das Vias Aéreas/imunologia , Asma Ocupacional/induzido quimicamente , Asma Ocupacional/enzimologia , Asma Ocupacional/imunologia , Estudos de Casos e Controles , Linhagem Celular Tumoral , Relação Dose-Resposta Imunológica , Feminino , Humanos , Interleucina-8/sangue , Interleucina-8/imunologia , Queratina-19/química , Masculino , Pessoa de Meia-Idade , Albumina Sérica/química , Inibidor Tecidual de Metaloproteinase-1/sangue , Inibidor Tecidual de Metaloproteinase-1/imunologia , Tolueno 2,4-Di-Isocianato/química , Tolueno 2,4-Di-Isocianato/imunologia , Fator de Crescimento Transformador beta1/sangue , Fator de Crescimento Transformador beta1/imunologia , Transglutaminases/química , Transglutaminases/imunologiaRESUMO
Glycopyrronium bromide (GB) is a muscarinic receptor antagonist that has been used as a long-acting bronchodilator in chronic obstructive pulmonary disease (COPD) patients. The aim of this study was to investigate the anti-inflammatory activity of inhaled GB in a cigarette smoke-induced acute lung inflammation mouse model. We found that aerosol pre-treatment with GB suppresses the accumulation of neutrophils and macrophages in the bronchoalveolar lavage fluid (BALF) in cigarette smoke (CS)-exposed mice. GB at doses of 300 and 600 µg/ml significantly inhibited the CS-induced increases in the mRNA and protein expression levels of interleukin (IL)-1ß, tumor necrosis factor (TNF)-α, monocyte chemotactic protein (MCP)-1 and transforming growth factor (TGF)-ß1 in lung tissues and the BALF. Moreover, GB at a dose of 600 µg/ml significantly inhibited the CS-induced changes in glutathione (GSH) and myeloperoxidase (MPO) activities in the BALF, decreased the CS-induced expression of matrix metalloproteinases (MMP)-9, and increased the CS-induced expression of tissue inhibitor of metalloproteinases (TIMP)-1, as determined through the immunohistochemical staining of lung tissue. Our results demonstrate the beneficial effects of inhaled GB on the inflammatory reaction in COPD.
Assuntos
Anti-Inflamatórios/uso terapêutico , Glicopirrolato/uso terapêutico , Antagonistas Muscarínicos/uso terapêutico , Pneumonia/tratamento farmacológico , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Animais , Anti-Inflamatórios/farmacologia , Líquido da Lavagem Broncoalveolar/citologia , Citocinas/genética , Citocinas/imunologia , Modelos Animais de Doenças , Feminino , Glicopirrolato/farmacologia , Linfócitos/imunologia , Macrófagos/imunologia , Metaloproteinase 9 da Matriz/imunologia , Camundongos , Camundongos Endogâmicos ICR , Antagonistas Muscarínicos/farmacologia , Neutrófilos/imunologia , Estresse Oxidativo/efeitos dos fármacos , Pneumonia/induzido quimicamente , Pneumonia/imunologia , Doença Pulmonar Obstrutiva Crônica/imunologia , Fumaça , Inibidor Tecidual de Metaloproteinase-1/imunologia , NicotianaRESUMO
The tissue inhibitors of metalloproteinases (TIMPs) are well recognized for their role in extracellular matrix remodeling by controlling the activity of matrix metalloproteinases (MMPs). Independent of MMP inhibition, TIMPs act as signaling molecules with cytokine-like activities thereby influencing various biological processes including cell growth, apoptosis, differentiation, angiogenesis, and oncogenesis. Recent studies on TIMP-1's cytokine functions have identified complex regulatory networks involving a specific surface receptor and subsequent signaling pathways including miRNA-mediated posttranscriptional regulation of gene expression that ultimately control the fate and behavior of the cells. The present review summarizes the current knowledge on TIMP-1 as a cytokine modulator of cell functions, outlines recent progress in defining molecular pathways that transmit TIMP-1 signals from the cell periphery into the nucleus, and discusses TIMP-1's role as a cytokine in the pathophysiology of cancer and other human diseases.