RESUMEN
OBJECTIVE: High-grade neuroendocrine cervical cancer (HGNECC) is a rare and aggressive cervical cancer subtype. In this study, we aimed to evaluate the prognostic value of fluorodeoxyglucose-PET/computed tomography (CT) parameters for HGNECC. MATERIALS AND METHODS: This single-center retrospective study included 29 patients with HGNECC who underwent fluorodeoxyglucose-PET/CT scan followed by surgery between 2006 and 2016. RESULTS: The median follow-up period was 40 (range, 4-184) months. After surgery, the resection margins were tumor-negative in 28 patients (96.6%), 8 (27.6%) patients had parametrial tumor invasion, and 7 patients (24.1%) tested positive for lymph node metastasis. The tumor recurred in 20 patients (69%) and 18 patients (62.1%) died during the observation period. In the univariate analyses, age and total lesion glycolysis (TLG) were associated with worse disease-free survival (DFS) (age, hazard ratio 1.056, 95% CI 1.014-1.100, P â =â 0.009; TLG2.5, hazard ratio 1.003, 95% CI 1-1.006, P â =â 0.033; and TLG3.0, hazard ratio 1.003, 95% CI 1-1.006, P â =â 0.034). In the multivariate analyses, older age and higher TLG3.0 were identified as independent poor prognostic factors for DFS (age, hazard ratio 1.058, 95% CI 1.014-1.104, P â =â 0.009; TLG3.0, hazard ratio 1.004, 95% CI 1-1.007, P â =â 0.033), while resection margin involvement was identified as an independent factor to predict poor overall survival (hazard ratio 20.717, 95% CI 1.289-332.964, P â =â 0.032). CONCLUSION: Among the preoperative fluorodeoxyglucose-PET/CT parameters, TLG3.0 may be useful for predicting DFS in patients with HGNECC.
RESUMEN
PURPOSE: Diesel exhaust particles (DEPs) can induce and trigger airway hyperresponsiveness (AHR) and inflammation. The aim of this study was to investigate the effect of long-term DEP exposure on AHR, inflammation, lung fibrosis, and goblet cell hyperplasia in a mouse model. METHODS: BALB/c mice were exposed to DEPs 1 hour a day for 5 days a week for 3 months in a closed-system chamber attached to a ultrasonic nebulizer (low dose: 100 µg/m³ DEPs, high dose: 3 mg/m³ DEPs). The control group was exposed to saline. Enhanced pause was measured as an indicator of AHR. Animals were subjected to whole-body plethysmography and then sacrificed to determine the performance of bronchoalveolar lavage and histology. RESULTS: AHR was higher in the DEP group than in the control group, and higher in the high-dose DEP than in the low-dose DEP groups at 4, 8, and 12 weeks. The numbers of neutrophils and lymphocytes were higher in the high-dose DEP group than in the low-dose DEP group and control group at 4, 8, and 12 weeks. The levels of interleukin (IL)-5, IL-13, and interferon-γ were higher in the low-dose DEP group than in the control group at 12 weeks. The level of IL-10 was higher in the high-dose DEP group than in the control group at 12 weeks. The level of vascular endothelial growth factor was higher in the low-dose and high-dose DEP groups than in the control group at 12 weeks. The level of IL-6 was higher in the low-dose DEP group than in the control group at 12 weeks. The level of transforming growth factor-ß was higher in the high-dose DEP group than in the control group at 4, 8, and 12 weeks. The collagen content and lung fibrosis in lung tissue was higher in the high-dose DEP group at 8 and 12 weeks. CONCLUSIONS: These results suggest that long-term DEP exposure may increase AHR, inflammation, lung fibrosis, and goblet cell hyperplasia in a mouse model.
RESUMEN
Peroxisome proliferator-activated receptor gamma coactivator 1 beta (PPARGC1B) is a coactivator of estrogen receptor (ER)α and ERß. We previously demonstrated a significant association between a variant of exon 5 of the PPARGC1B gene (+102525 G>A, R265Q) and airway hyperreactivity (AHR). The aims of the study were to evaluate the genetic effects of variants of the PPARGC1B gene on the function of ERs. PPARGC1B +102525G and A gene constructs were generated using PCR and cloned into a pCMV4 promoter vector. A luciferase reporter assay was undertaken in 293T cells cotransfected with one of the PPARGC1B +102525G>A constructs, ERα, and an estrogen response element (ERE) containing a luciferase construct after treatment with 17ß-estradiol. According to the luciferase reporter assay, the +102525A allele showed higher ERα activity than the +102525G allele in response to stimulation with 17ß-estradiol. In addition, the interaction between ERα and PPARGC1B was evaluated by coprecipitation assay. Human influenza hemagglutinin-tagged PPARGC1B coprecipitated more intensely with ERα in the +102525A than the +102525G construct after 17ß estradiol treatment. The variant +102525A allele enhances the activity of ERα to a greater degree than the +102525G allele of PPARGC1B.
Asunto(s)
Proteínas Portadoras/genética , Estradiol/farmacología , Receptor alfa de Estrógeno/genética , Receptor alfa de Estrógeno/metabolismo , Exones/genética , Receptores de Estrógenos/metabolismo , Proteínas Portadoras/metabolismo , Células Cultivadas , Estrógenos/metabolismo , Humanos , PPAR gamma/metabolismo , Regiones Promotoras Genéticas , ARN Mensajero , Proteínas de Unión al ARN , Activación Transcripcional/efectos de los fármacos , Activación Transcripcional/genéticaRESUMEN
Chronic microglial activation endangers neuronal survival through the release of various toxic pro-inflammatory molecules; thus, negative regulators of microglial activation have been identified as potential therapeutic candidates for several neurological diseases. In this study, we investigated the inhibitory effects of an ethanol extract of Cnidium officinale rhizomes (EECO), which has been used as a herbal drug in Oriental medicine, on the production of lipopolysaccharide (LPS)-induced pro-inflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), as well as that of pro-inflammatory cytokines in BV2 microglia cells. EECO significantly inhibited the excess production of NO and PGE2 in LPS-stimulated BV2 microglia cells. It also attenuated the expression of inducible NO synthase, cyclooxygenase-2, as well as that of pro-inflammatory cytokines, such as interleukin-1ß and tumor necrosis factor-α. Moreover, EECO exhibited anti-inflammatory properties by suppressing nuclear factor-κB (NF-κB) translocation and the activation of the phosphoinositide 3-kinase/Akt pathway in LPS-stimulated BV2 cells. These results indicate that EECO exerts anti-inflammatory effects in LPS-stimulated BV2 microglial cells by inhibiting pro-inflammatory mediators and cytokine production by blocking the NF-κB pathway. These findings suggest that EECO has substantial therapeutic potential for the treatment of neurodegenerative diseases accompanied by microglial activation.
Asunto(s)
Antiinflamatorios/farmacología , Cnidium/química , Microglía/efectos de los fármacos , Extractos Vegetales/farmacología , Transducción de Señal , Animales , Línea Celular , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/metabolismo , Etanol , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Lipopolisacáridos/efectos adversos , Ratones , Microglía/citología , FN-kappa B/antagonistas & inhibidores , FN-kappa B/genética , FN-kappa B/metabolismo , Óxido Nítrico/biosíntesis , Óxido Nítrico Sintasa de Tipo II/genética , Óxido Nítrico Sintasa de Tipo II/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Fosfatidilinositol 3-Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Proteínas Proto-Oncogénicas c-akt/metabolismo , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Dendropanax morbifera Leveille is found throughout southwestern Korea, and has been used in traditional medicine for various diseases, such as migraine headache, infectious diseases, skin diseases and dysmenorrhea. However, the molecular mechanisms of D. morbifera concerning its biochemical actions in cancer have not yet been clearly elucidated. In the present study, we investigated the pro-apoptotic effects of an ethanol extract of D. morbifera stem bark (EEDM) on human leukemia U937 cells. EEDM markedly inhibited the growth of U937 cells by decreasing cell proliferation and inducing apoptosis. EEDM-induced apoptosis in U937 cells was associated with the upregulation of death receptor-related protein levels and downregulation of anti-apoptotic IAP family proteins. The increase in apoptosis was also associated with proteolytic activation of caspase-8, -9 and -3, inhibition of antiapoptotic Bcl-2 and Bcl-xL expression, Bid cleavage, and loss of MMP suggesting that apoptosis of U937 cells induced by EEDM was through the extrinsic and intrinsic pathways. However, a pan-caspase inhibitor, z-VED-fmk, significantly inhibited EEDM-induced U937 cell apoptosis indicating that the caspases were key regulators of apoptosis in response to EEDM in U937 cells. Our data suggest that D. morbifera may be a potential anticancer agent for cancer treatment.
Asunto(s)
Apoptosis/efectos de los fármacos , Araliaceae/química , Caspasas/metabolismo , Leucemia/patología , Extractos Vegetales/farmacología , Transducción de Señal/efectos de los fármacos , Western Blotting , Ciclo Celular/efectos de los fármacos , Permeabilidad de la Membrana Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Fibroblastos/citología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Citometría de Flujo , Humanos , Leucemia/tratamiento farmacológico , Leucemia/metabolismo , Hígado/citología , Hígado/efectos de los fármacos , Hígado/metabolismo , Pulmón/citología , Pulmón/efectos de los fármacos , Pulmón/metabolismo , Potencial de la Membrana Mitocondrial/efectos de los fármacos , Células U937 , Ensayos Antitumor por Modelo de XenoinjertoRESUMEN
PURPOSE: This study was done to identify the parenting experience of mothers of premature infants in order to provide basic data for educational solutions and desirable directions. METHODS: Q-methodology was used as it provides a method of analyzing the subjectivity of each item. The participants were 33 mothers of premature infants who sorted 34 selected Q-statements which were then classified into the shape of a normal distribution using a 9-point scale. Subjectivity on parenting experience among the mothers was analyzed using the pc-QUANAL program. RESULTS: Four types of parenting experience were identified. Type I was named 'struggling', type II, 'self blame', type III, 'information collecting', and type IV, 'self-introspection'. CONCLUSION: The results of this study indicate that different approaches to educational programs are needed for mothers of premature infants based on the four types of parenting experience.
Asunto(s)
Recien Nacido Prematuro , Madres/psicología , Responsabilidad Parental , Adulto , Demografía , Femenino , Humanos , Recién Nacido , Entrevistas como Asunto , Q-Sort , Encuestas y CuestionariosRESUMEN
For the past two decades, a huge number of genetic studies have been conducted to identify the genetic variants responsible for asthma risk. Several types of genetic and genomic approaches, including linkage analysis, candidate gene single nucleotide polymorphism studies, and whole genome-wide association studies have been applied. In this review article, the results of these approaches are summarized, and their limitations are discussed. Additionally, perspectives for applying upcoming new epigenetic or genomic technologies, such as copy number variation, are introduced to increase our understanding of new omic approaches to asthma genetics.
RESUMEN
PURPOSE: Severe asthma is characterized by high medication requirements to maintain good disease control or by persistent symptoms despite high medication use. The transfer of bone marrow-derived mesenchymal stem cells (BMDMSCs) to the injured lungs is a possible treatment for severe asthma. This study investigated the therapeutic effects of BMDMSCs in airway remodeling and inflammation in an experimental toluene diisocyanate (TDI)-induced asthma animal model of severe asthma. METHODS: BMDMSCs were transferred into rats after TDI inhalation. Bronchoalveolar lavage (BAL) cell profiles, histological changes including an inflammatory index and goblet cell hyperplasia, and the airway response to methacholine using plethysmography were analyzed. Smooth muscle actin (SMA) and proliferating cell nuclear antigen (PCNA) protein expression were observed in lung tissue using immunohistochemical staining. The collagen content was measured in lung tissue sections and lung extracts using Masson's trichrome staining and an immunoassay kit. RESULTS: The numbers of inflammatory cells in BAL fluid, histological inflammatory index, airway response to methacholine, number of goblet cells, and amount of collagen were increased in TDI-treated rats compared with sham rats (P=0.05-0.002). BMDMSC transfer significantly reduced the TDI-induced increase in the inflammatory index and numbers of eosinophils and neutrophils in BAL fluid to levels seen in sham-treated rats (P<0.05). BMDMSC transfer significantly reduced the number of goblet cells, collagen deposition, and immune staining for SMA and PCNA with concomitant normalization of the airway response to methacholine. CONCLUSIONS: The systemic transfer of BMDMSCs effectively reduced experimental TDI-induced airway inflammation and remodeling and airway hyperreactivity.
RESUMEN
BACKGROUND: Interleukin-8 (IL-8) is a potent chemo-attractant cytokine responsible for neutrophil infiltration in lungs with idiopathic pulmonary fibrosis (IPF). The IL-8 protein and mRNA expression are increased in the lung with IPF. We evaluated the effect of single nucleotide polymorphisms (SNPs) of the IL-8 gene on the risk of IPF. METHODS: One promoter (rs4073T>A) and two intronic SNPs (rs2227307T>G and rs2227306C>T) of the IL-8 genes were genotyped in 237 subjects with IPF and 456 normal controls. Logistic regression analysis was applied to evaluate the association of these SNPs with IPF. IL-8 in BAL fluids was measured using a quantitative sandwich enzyme immunoassay, and promoter activity was assessed using the luciferase reporter assay. RESULTS: The minor allele frequencies of rs4073T>A and rs2227307T>G were significantly lower in the 162 subjects with surgical biopsy-proven IPF and 75 subjects with clinical IPF compared with normal controls in the recessive model (OR = 0.46 and 0.48, p = 0.006 and 0.007, respectively). The IL-8 protein concentration in BAL fluids significantly increased in 24 subjects with IPF compared with 14 controls (p = 0.009). Nine IPF subjects homozygous for the rs4073 T>A common allele exhibited higher levels of the IL-8 protein compared with six subjects homozygous for the minor allele (p = 0.024). The luciferase activity of the rs4073T>A common allele was significantly higher than that of the rs4073T>A minor allele (p = 0.002). CONCLUSION: The common allele of a promoter SNP, rs4073T>A, may increase susceptibility to the development of IPF via up-regulation of IL-8.
Asunto(s)
Fibrosis Pulmonar Idiopática/genética , Interleucina-8/genética , Polimorfismo de Nucleótido Simple , Regiones Promotoras Genéticas , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Líquido del Lavado Bronquioalveolar/inmunología , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Ensayo de Inmunoadsorción Enzimática , Femenino , Frecuencia de los Genes , Genes Reporteros , Predisposición Genética a la Enfermedad , Células HEK293 , Homocigoto , Humanos , Fibrosis Pulmonar Idiopática/diagnóstico , Fibrosis Pulmonar Idiopática/inmunología , Interleucina-8/metabolismo , Intrones , Modelos Logísticos , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Fenotipo , Modelos de Riesgos Proporcionales , República de Corea , Medición de Riesgo , Factores de Riesgo , Transfección , Regulación hacia ArribaRESUMEN
BACKGROUND: Signal-regulated palmitoylation of RGS7BP(regulator of G-protein-signaling 7-binding protein) initiates the activation of G-protein-coupled receptors (GPCRs), including muscarinic receptors, which contribute to the development of asthma and its subphenotypes. OBJECTIVE: To determine the association of RGS7BP gene polymorphisms with the development of aspirin-exacerbated respiratory disease (AERD). METHODS: We evaluated the association of RGS7BP gene polymorphisms with response to oral aspirin challenge and with responsiveness to methacholine challenge. RGS7BP messenger RNA splice variants in peripheral blood platelets from patients with different single-nucleotide polymorphisms were analyzed by reverse-transcription polymerase chain reaction. RESULTS: Logistic regression analysis of RGS7BP gene polymorphisms in patients with AERD (n = 102) and aspirin-tolerant asthma (n = 429) revealed that a haplotype of block 3 consisting of rare alleles +98092 C>G, +98853 C>T, and +104450 T>G of the RGS7BP gene was associated with AERD. Multiple linear regression analysis showed that asthmatic patients carrying ht2/ht2 in block 3 were more responsive to aspirin challenge than those not carrying ht2 (P = .008 in a codominant model). The log-transformed provocation concentration that caused a decrease in forced expiratory volume in 1 second of 20% for methacholine was significantly dependent on the BL3-ht2 haplotype. No significant differences in platelet expression of different RGS7BP messenger RNA splice variants were detected between those with and without the BL3-ht2 haplotype. CONCLUSION: BL3-ht2 of RGS7BP may be an important genetic variant associated with AERD. The haplotype of block 3 may play a protective role against aspirin hypersensitivity in asthma, perhaps by altering the responsiveness of muscarinic receptors.
Asunto(s)
Aspirina/efectos adversos , Asma/inducido químicamente , Asma/genética , Proteínas Portadoras/genética , Hipersensibilidad a las Drogas/genética , Receptores Acoplados a Proteínas G/genética , Adolescente , Adulto , Anciano , Aspirina/inmunología , Asma/inmunología , Proteínas Portadoras/inmunología , Hipersensibilidad a las Drogas/etiología , Hipersensibilidad a las Drogas/inmunología , Femenino , Volumen Espiratorio Forzado , Genotipo , Humanos , Péptidos y Proteínas de Señalización Intracelular , Modelos Lineales , Desequilibrio de Ligamiento , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Proteínas RGS , ARN/química , ARN/genética , Receptores Acoplados a Proteínas G/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Adulto JovenRESUMEN
Asthma manifests as TH2-dominant airway inflammation regulated by inducible T-cell kinase (ITK). To investigate associations between genetic variants of the ITK gene and asthma, 31 single-nucleotide polymorphisms (SNPs) were genotyped in 303 normal controls and 498 asthmatics and the two groups were compared using logistic regression models. The functional effects of the ITK promoter SNP were assessed using pGL3 luciferase reporter systems and gel-shift assays. The minor allele-196C>T in the promoter region of the ITK gene was significantly more frequent in asthmatics than in controls. The luciferase activity of the PGL3-ITK-196T allele construct was higher than that of the -196C allele. In the gel-shift assay, -196T double-stranded oligonucleotides bound more strongly to Jurkat cell nuclear protein compared to the -196C double-stranded oligonucleotides. People with the -rare allele 196C>T may be more susceptible to asthma via transcriptional regulation of the ITK gene.
Asunto(s)
Asma/genética , Polimorfismo de Nucleótido Simple , Proteínas Tirosina Quinasas/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Regiones Promotoras Genéticas , Adulto JovenRESUMEN
RATIONALE: Airway inflammation and remodeling during asthma are attributed to the altered expression of biologically relevant proteins. OBJECTIVES: To search for asthma-specific proteins in bronchoalveolar lavage fluid (BAL) from individuals with asthma and to validate the identified proteins in an experimental model of asthma. METHODS: Liquid chromatography-tandem mass spectrometry was performed to identify proteins in BAL fluid found by two dimensional electrophoresis (2DE) to be differentially expressed in subjects with asthma versus control subjects. Group-specific component (Gc) and mRNA levels were measured using an ELISA, Western blots, and PCR. A neutralization study using an antibody against Gc protein was performed in an experimental asthma model. MEASUREMENTS AND MAIN RESULTS: Based on 2DE, 15 proteins were significantly up-regulated or down-regulated in eight subjects with asthma compared with eight control subjects. The protein levels of Gc, hemopexin, and haptoglobin-b were increased, whereas the a1- antitrypsin and glutathione S-transferase levels were decreased in subjects with asthma. The Gc concentration in BAL fluid was significantly elevated in 67 subjects with asthma compared with that in 22 control subjects (P < 0.009). The Gc was significantly correlated with the neutrophil percentage in BAL fluid of subjects with asthma (P = 0.001). Gc mRNA and protein levels were higher in ovalbumin-sensitized/ challenged asthma mice than in sham-treated mice. Gc protein were expressed on alveolar macrophages and on epithelial cells. Treatment with an anti-Gc antibody dose-dependently reduced the ovalbumin sensitization/challenge-induced enhancement of airway hyperreactivity, airway inflammation, goblet cell hyperplasia,and levels of eotaxin, interleukin-4, -5, and -13, and interferon-g. CONCLUSIONS: Gc may be involved in the development of asthma, and the neutralization of Gc protein could be a therapeutic strategy for asthma.
Asunto(s)
Asma/metabolismo , Líquido del Lavado Bronquioalveolar/química , Proteínas/metabolismo , Adulto , Animales , Asma/etiología , Asma/fisiopatología , Western Blotting , Cromatografía Liquida , Ensayo de Inmunoadsorción Enzimática , Femenino , Estudios de Seguimiento , Glutatión Transferasa/genética , Glutatión Transferasa/metabolismo , Humanos , Interleucina-4/genética , Interleucina-4/metabolismo , Masculino , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Proteínas/genética , ARN Mensajero/análisis , Espectrometría de Masas en TándemRESUMEN
AIMS: Cysteinyl leukotrienes are inactivated by acetyl coenzyme A-dependent N-acetyltransferase (NAT). Thus, functional alterations of the NAT gene may contribute to the risk of aspirin-intolerant asthma. MATERIALS & METHODS: Asthmatics (n = 438) were categorized into aspirin-intolerant asthma (15% or greater decrease in the forced expiratory volume in 1 s or cutaneous reactions, n = 170) or aspirin-tolerant asthma (n = 268) groups. In total, 14 polymorphisms of the NAT2 gene were genotyped by a single-base extension method. RESULTS: The distributions of all loci of the 14 SNPs were in Hardy-Weinberg equilibrium (p > 0.05). Among the 14 SNPs, six common SNPs (minor allele frequency >5%) in a Korean population were used for haplotype construction and further statistical analysis. The logistic regression analysis demonstrated that NAT2 -9246G>C and haplotype 2 (TCACGG) were significantly associated with the risk of aspirin-intolerant asthma. The rare allele frequencies of the SNP and Ht2 were significantly higher in the aspirin-intolerant asthma group than in the aspirin-tolerant asthma group (p(corr) = 0.03 and p(corr) = 0.02 in codominant model). CONCLUSION: In a large genetic epidemiology study of aspirin-intolerant asthma in a Korean population, genetic polymorphisms of NAT2 were found to be related to a risk of aspirin hypersensitivity among asthmatics.
Asunto(s)
Arilamina N-Acetiltransferasa/genética , Aspirina/efectos adversos , Asma/genética , Hipersensibilidad a las Drogas/genética , Polimorfismo Genético , Adulto , Alelos , Pueblo Asiatico/genética , Aspirina/inmunología , Asma/fisiopatología , Estudios de Casos y Controles , Cisteína/inmunología , Tolerancia a Medicamentos/genética , Femenino , Frecuencia de los Genes , Genotipo , Haplotipos , Humanos , Corea (Geográfico) , Leucotrienos/inmunología , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido SimpleRESUMEN
Colony-stimulating factor 1 receptor (CSF1R) is expressed in monocytes/macrophages and dendritic cells. These cells play important roles in the innate immune response, which is regarded as an important aspect of asthma development. Genetic alterations in the CSF1R gene may contribute to the development of asthma. We investigated whether CSF1R gene polymorphisms were associated with the risk of asthma. Through direct DNA sequencing of the CSF1R gene, we identified 28 single nucleotide polymorphisms (SNPs) and genotyped them in 303 normal controls and 498 asthmatic patients. Expression of CSF1R protein and mRNA were measured on CD14-positive monocytes and neutrophils in peripheral blood of asthmatic patients using flow cytometry and real-time PCR. Among the 28 polymorphisms, two intronic polymorphism (+20511C>T and +22693T>C) were associated with the risk of asthma by logistic regression analysis. The frequencies of the minor allele at CSF1R +20511C>T and +22693T>C were higher in asthmatic subjects than in normal controls (4.6 vs. 7.7%, p = 0.001 in co-dominant and dominant models; 16.4 vs. 25.8%, p = 0.0006 in a recessive model). CSF1R mRNA levels in neutrophils of the asthmatic patients having the +22693CC allele were higher than in those having the +22693TT allele (p = 0.026). Asthmatic patients with the +22693CC allele also showed significantly higher CSF1R expression on CD14-positive monocytes and neutrophils than did those with the +22693TT allele (p = 0.045 and p = 0.044). The +20511C>T SNP had no association with CSF1R mRNA or protein expression. In conclusion, the minor allele at CSF1R +22693T>C may have a susceptibility effect in the development of asthma, via increased CSF1R protein and mRNA expression in inflammatory cells.
Asunto(s)
Asma/genética , Polimorfismo de Nucleótido Simple , Receptor de Factor Estimulante de Colonias de Macrófagos/genética , Alelos , Asma/etiología , Asma/inmunología , Secuencia de Bases , Estudios de Casos y Controles , Cartilla de ADN/genética , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Humanos , Inmunidad Innata/genética , Desequilibrio de Ligamiento , Modelos Genéticos , Monocitos/inmunología , Neutrófilos/inmunología , ARN Mensajero/sangre , ARN Mensajero/genética , Receptor de Factor Estimulante de Colonias de Macrófagos/sangreRESUMEN
RATIONALE: Idiopathic pulmonary fibrosis (IPF) is caused by alterations in expression of proteins involved in multiple pathways, including matrix deposition, inflammation, injury, and repair. OBJECTIVES: To understand the pathogenic changes in lung protein expression in IPF and to evaluate apolipoprotein (Apo) A-I as a candidate therapeutic molecule. METHODS: Two-dimensional electrophoresis was adopted for differential display proteomics. Reverse-transcriptase polymerase chain reaction, Western blotting, immunohistochemical staining, and ELISA were performed for identification and quantitative measurement of Apo A-I in bronchoalveolar lavage fluids from subjects with IPF and experimental bleomycin-induced mice. MEASUREMENTS AND MAIN RESULTS: Sixteen protein spots showed differences in relative intensity between IPF (n = 14) and healthy control subjects (n = 8). Nano liquid chromatography-tandem mass spectrometry (LC-MS/MS) revealed increase of haptoglobulin and decrease of alpha(1)-antitrypsin, alpha(1)-antichymotrypsin, macrophage capping protein, angiotensinogen, hemoglobin chain B, Apo A-I, clusterin, protein disulfide isomerase A3, immunoglobulin, and complement C4A in IPF compared with normal control subjects (P = 0.006-0.044). Apo A-I concentrations were lower in bronchoalveolar lavage fluids from subjects with IPF (n = 28) than in normal control subjects (n = 18; P < 0.01). In bleomycin-treated mice, Apo A-I protein in BALF was lower than that in sham-treated control animals. Immunohistochemical analysis showed positive staining on intraalveolar macrophages and epithelial cells of the lungs. Intranasal treatment with Apo A-I protein reduced the bleomycin-induced increases in number of inflammatory cells and collagen deposition in sham-treated mice in a dose-dependent manner. CONCLUSIONS: Alterations of several inflammatory and antiinflammatory proteins in the lungs may be related to the pathogenesis of IPF, and local treatment with Apo A-I is very effective against the development of experimental lung injury and fibrosis.
Asunto(s)
Apolipoproteína A-I/biosíntesis , Apolipoproteína A-I/uso terapéutico , Fibrosis Pulmonar Idiopática/tratamiento farmacológico , Fibrosis Pulmonar Idiopática/metabolismo , Anciano , Animales , Apolipoproteína A-I/genética , Apolipoproteína A-I/metabolismo , Bleomicina , Western Blotting , Líquido del Lavado Bronquioalveolar/química , Modelos Animales de Enfermedad , Electroforesis en Gel Bidimensional , Ensayo de Inmunoadsorción Enzimática , Femenino , Células Espumosas/metabolismo , Células Espumosas/patología , Humanos , Fibrosis Pulmonar Idiopática/patología , Fibrosis Pulmonar Idiopática/fisiopatología , Pulmón/metabolismo , Pulmón/patología , Pulmón/fisiopatología , Masculino , Ratones , Persona de Mediana Edad , Proteómica/métodos , Fibrosis Pulmonar/inducido químicamente , Fibrosis Pulmonar/tratamiento farmacológico , Fibrosis Pulmonar/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: No effective treatment for acute lung injury and fibrosis currently exists. Aim of this study was to investigate the time-dependent effect of bone marrow-derived mesenchymal stem cells (BMDMSCs) on bleomycin (BLM)-induced acute lung injury and fibrosis and nitric oxide metabolites and inflammatory cytokine production. METHODS: BMDMSCs were transferred 4 days after BLM inhalation. Wet/dry ratio, bronchoalveolar lavage cell profiles, histologic changes and deposition of collagen were analyzed. RESULTS: Nitrite, nitrate and cytokines were measured weekly through day 28. At day 7, the wet/dry ratio, neutrophilic inflammation, and amount of collagen were elevated in BLM-treated rats compared to sham rats (p = 0.05-0.002). Levels nitrite, nitrate, IL-1beta, IL-6, TNF-alpha, TGF-beta and VEGF were also higher at day 7 (p < 0.05). Degree of lymphocyte and macrophage infiltration increased steadily over time. BMDMSC transfer significantly reduced the BLM-induced increase in wet/dry ratio, degree of neutrophilic infiltration, collagen deposition, and levels of the cytokines, nitrite, and nitrate to those in sham-treated rats (p < 0.05). Fluorescence in situ hybridization localized the engrafted cells to areas of lung injury. CONCLUSION: Systemic transfer of BMDMSCs effectively reduced the BLM-induced lung injury and fibrosis through the down-regulation of nitric oxide metabolites, and proinflammatory and angiogenic cytokines.
Asunto(s)
Citocinas/metabolismo , Lesión Pulmonar/metabolismo , Lesión Pulmonar/cirugía , Trasplante de Células Madre Mesenquimatosas , Óxido Nítrico/metabolismo , Fibrosis Pulmonar/metabolismo , Fibrosis Pulmonar/cirugía , Animales , Bleomicina , Células Cultivadas , Lesión Pulmonar/inducido químicamente , Masculino , Fibrosis Pulmonar/inducido químicamente , Ratas , Ratas Sprague-DawleyRESUMEN
BLT2 is a low-affinity receptor for leukotriene B(4) (LTB(4)), a potent lipid mediator of inflammation generated from arachidonic acid via the 5-lipoxygenase pathway. Unlike BLT1, a high-affinity receptor for LTB(4), no clear physiological function has yet been identified for BLT2, especially with regard to the pathogenesis of asthma. The aim of this study was to investigate whether BLT2 plays a role in the pathogenesis of asthma. A murine model of allergic asthma was used to evaluate the role of BLT2 in ovalbumin-induced airway inflammation and airway hyperresponsiveness. The levels of BLT2 mRNA and its ligand, LTB(4), in the lung airway were highly elevated after ovalbumin challenge, and down-regulation of BLT2 with antisense BLT2 oligonucleotides markedly attenuated airway inflammation and airway hyperresponsiveness. Further analysis, aimed at identifying mediators downstream of BLT2, revealed that BLT2 activation led to elevation of reactive oxygen species and subsequent activation of NF-kappaB, thus inducing the expression of vascular cell adhesion molecule-1, which is known to be involved in eosinophil infiltration into the lung airway. Together, our results suggest that BLT2 plays a pivotal, mediatory role in the pathogenesis of asthma, acting through a "reactive oxygen species-NF-kappaB"-linked inflammatory signaling pathway.
Asunto(s)
Hiperreactividad Bronquial/complicaciones , Neumonía/complicaciones , Receptores de Leucotrieno B4/antagonistas & inhibidores , Receptores de Leucotrieno B4/metabolismo , Animales , Asma/genética , Asma/fisiopatología , Biopsia , Bronquios/efectos de los fármacos , Bronquios/patología , Hiperreactividad Bronquial/patología , Hiperreactividad Bronquial/fisiopatología , Movimiento Celular/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Pulmón/efectos de los fármacos , Pulmón/patología , Pulmón/fisiopatología , Ratones , FN-kappa B/metabolismo , Ovalbúmina , Neumonía/patología , Neumonía/fisiopatología , ARN sin Sentido/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptores de Leucotrieno B4/genética , Transducción de Señal/efectos de los fármacos , Linfocitos T/citología , Linfocitos T/efectos de los fármacos , Tetrazoles/farmacología , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
Cordyceps militaris is well known as a traditional medicinal mushroom and is a potentially interesting candidate for use in cancer treatment. In this study, the potential of the water extract of C. militaris (WECM) to induce apoptosis in human lung carcinoma A549 cells and its effects on telomerase activity were investigated. The growth inhibition and apoptosis induction by WECM treatment in A549 cells was associated with the induction of Fas, catalytic activation of caspase-8, and Bid cleavage. Activation of caspases, downregulation of anti-apoptotic Bcl-2 expression, and upregulation of pro-apoptotic Bax protein were also observed in WECM-treated cells. However, the cytotoxic effects and apoptotic characteristics induced by WECM were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrates the important role that caspase-3 plays in the process. In addition, WECM exerted a dose-dependent inhibition of telomerase activity via downregulation of human telomerase reverse transcriptase (hTERT), c-myc and Sp1 expression. Taken together, the data from this study indicate that WECM induces the apoptosis of A549 cells through a signaling cascade of death receptor-mediated extrinsic and mitochondria-mediated intrinsic caspase pathways. It was also conclude that apoptotic events due to WECM were mediated with diminished telomerase activity through the inhibition of hTERT transcriptional activity.
Asunto(s)
Antineoplásicos Fitogénicos/farmacología , Apoptosis/efectos de los fármacos , Cordyceps/química , Neoplasias Pulmonares/enzimología , Neoplasias Pulmonares/patología , Telomerasa/antagonistas & inhibidores , Antineoplásicos Fitogénicos/química , Western Blotting , Caspasas/metabolismo , Línea Celular Tumoral , Núcleo Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Fragmentación del ADN/efectos de los fármacos , Electroforesis en Gel de Agar , Activación Enzimática/efectos de los fármacos , Citometría de Flujo , Humanos , Extractos Vegetales/química , Extractos Vegetales/farmacología , ARN/biosíntesis , ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Solventes , Telomerasa/metabolismo , AguaRESUMEN
Anthocyanins are a class of flavonoids, widely spread throughout the plant kingdom, that exhibit important anti-oxidant and anti-inflammatory actions as well as chemotherapeutic effects. However, little is known concerning the molecular mechanisms by which these activities are exerted. In this study, we investigated the anthocyanins isolated from Vitis coignetiae Pulliat for their potential anti-proliferative and apoptotic effects on human leukemia U937 cells. It was found that these anthocyanins inhibit cell viability and induce apoptotic cell death of U937 cells in a dose-dependent manner, as measured by hemocytometer counts, by alteration in the mitochondrial membrane potential, by increases in sub-G1 populations and by DNA ladder formation. Apoptosis of U937 cells by anthocyanins was associated with modulation of expression of Bcl-2 and IAP family members. Consequently, anthocyanin treatment induced proteolytic activation of caspase-3, -8 and -9, and a concomitant degradation of poly(ADP-ribose) polymerase. However, anthocyanin-induced growth inhibition and apoptosis were significantly attenuated in Bcl-2 overexpressing U937 cells. Furthermore, z-DEVD-fmk, a caspase-3 specific inhibitor, blocked apoptosis and increased the survival of anthocyanin-treated U937 cells. Taken together, these results show that Bcl-2 and caspases are key regulators of apoptosis in response to anthocyanins in human leukemia U937 cells.
Asunto(s)
Antocianinas/farmacología , Apoptosis , Caspasas/metabolismo , Regulación hacia Abajo , Regulación Enzimológica de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Supervivencia Celular , ADN/metabolismo , Activación Enzimática , Humanos , Potenciales de la Membrana , Membranas Mitocondriales/metabolismo , Células U937 , Vitis/metabolismoRESUMEN
RATIONALE: Gamma-secretase inhibitor (GSI) has been used to effectively block Notch signaling, which is implicated in the differentiation and functional regulation of T helper (Th) effector cells. In asthma, a subset of CD4(+) T cells is believed to initiate and perpetuate the disease. OBJECTIVES: The aim of this study was to evaluate the therapeutic potential of GSI against allergic asthma. METHODS: GSI was administered to an ovalbumin-sensitized mouse via an intranasal route at the time of ovalbumin challenge. MEASUREMENTS AND MAIN RESULTS: The administration of GSI inhibits asthma phenotypes, including eosinophilic airway inflammation, goblet cell metaplasia, methacholine-induced airway hyperresponsiveness, and serum IgE production. GSI treatment of bronchoalveolar lavage cells stimulated via TCR or non-TCR pathways led to a decrease in Th2 cytokine production with a concomitant increase in Th1 cytokine secretion. Expression of Hes-1, a target of Notch signaling, was down-regulated in conjunction with a reduction of Notch intracellular domain and GATA-3 levels after GSI treatment of bronchoalveolar lavage cells. GSI treatment resulted in an inhibition of NF-kappaB activation, and combined treatment with GSI and an NF-kappaB inhibitor augmented IFN-gamma production in a synergistic manner. CONCLUSIONS: These data suggest that GSI directly regulates Th1 and Th2 responses in allergic pulmonary inflammation through a Notch signaling-dependent pathway and that GSI is of high therapeutic value for treating asthma by inhibiting airway inflammatory responses.