RESUMO
The human paranasal sinuses are the major source of intrinsic nitric oxide (NO) production in the human airway. NO plays several roles in the maintenance of physiological homeostasis and the regulation of airway inflammation through the expression of three NO synthase (NOS) isoforms. Measuring NO levels can contribute to the diagnosis and assessment of allergic rhinitis (AR) and chronic rhinosinusitis (CRS). In symptomatic AR patients, pro-inflammatory cytokines upregulate the expression of inducible NOS (iNOS) in the inferior turbinate. Excessive amounts of NO cause oxidative damage to cellular components, leading to the deposition of cytotoxic substances. CRS phenotype and endotype classifications have provided insights into modern treatment strategies. Analyses of the production of sinus NO and its metabolites revealed pathobiological diversity that can be exploited for useful biomarkers. Measuring nasal NO based on different NOS activities is a potent tool for specific interventions targeting molecular pathways underlying CRS endotype-specific inflammation. We provide a comprehensive review of the functional diversity of NOS isoforms in the human sinonasal system in relation to these two major nasal disorders' pathologies. The regulatory mechanisms of NOS expression associated with the substrate bioavailability indicate the involvement of both type 1 and type 2 immune responses.
Assuntos
Mucosa Nasal/enzimologia , Óxido Nítrico Sintase/metabolismo , Óxido Nítrico/metabolismo , Seios Paranasais/enzimologia , Rinite Alérgica/fisiopatologia , Sinusite/fisiopatologia , Animais , Doença Crônica , Humanos , Isoenzimas , Rinite Alérgica/metabolismo , Sinusite/metabolismoRESUMO
The nasal tissues have the main consecutive roles of moistening and heating the air entering the respiratory tract and detecting odor via the activation of olfactory receptors in the neuro-olfactory epithelium. Initially, nasal toxicology was investigated to better assess the risk of nasal injuries caused by environmental toxicants or their active metabolites. Later, the characterization of the nasal toxicological barrier was a research concern for the purposes of intranasal drug delivery. Both fields allowed for an increase in our knowledge of the nasal xenobiotic-metabolizing enzymes and transporters that are highly expressed in this tissue. In addition to airborne toxicants or drugs, the main substrates for these proteins are natural volatiles known as odorants that emanate from our daily environment (food, perfume, plants, materials, congeners, etc.). Accordingly, another emerging field of interest has been developed that aims to understand the function of odorant-metabolizing enzymes (OMEs) in olfaction. Early in this field of research, OMEs were suspected to participate in the clearance of odorants from the receptor environment to avoid their saturation and thus maintain the sensitivity of neuronal detection. Other roles of OMEs that could significantly modulate olfaction were also considered, such as the involvement of odorant primary metabolites in the olfactory response. By combining enzymatic, physiological and sensory experimental approaches, recent advances have markedly improved our understanding of the contributions of OMEs to the olfactory process. This review combines recent data from the literature regarding nasal OME identification, localization, and activity and highlights the function of OMEs in olfaction.
Assuntos
Enzimas/metabolismo , Mucosa Nasal/metabolismo , Odorantes , Olfato/fisiologia , Animais , Humanos , Mucosa Nasal/enzimologia , Nariz/fisiologiaRESUMO
Chronic rhinosinusitis with nasal polyps (CRSwNP) is a chronic inflammatory disease of the paranasal sinuses, and its pathophysiology is not yet precisely known. It is suggested that oxygen free radicals play an important role in the pathogenesis of nasal polyposis. This study aimed to identify genetic polymorphisms of superoxide dismutase (SOD 2), catalase (CAT), and inducible nitric oxide synthase (iNOS) enzymes in eosinophilic CRSwNP and non-eosinophilic CRSwNP patients; the study also aimed to evaluate the effect of genetic polymorphism of antioxidant enzymes on CRSwNP etiopathogenesis. One hundred thirty patients, who received endoscopic sinus surgery due to CRSwNP, and 188 control individuals were included in this study. Nasal polyp tissues were divided into two groups histopathologically as eosinophilic CRSwNP and non-eosinophilic CRSwNP. Venous blood samples were taken from the patient and control groups. Polymorphisms in the Ala16Va1 gene, which is the most common variation of SOD-2 gene, and 21 A/T polymorphisms in catalase gene were evaluated with the restriction fragment length polymorphism method and -277 C/T polymorphism in the iNOS gene was evaluated with the DNA sequencing method. The GG genotype distribution for the (-277) A/G polymorphism in the iNOS gene was a statistically significant difference between eosinophilic CRSwNP and control groups (p < 0.05). The CC genotype distribution for the SOD2 A16V (C/T) polymorphism was not statistically significant in all groups (p > 0.05). The TT genotype distribution for the A/T polymorphism in catalase gene at position -21 was statistically significant differences in eosinophilic CRSwNP and control groups (p < 0.05). Increased free oxygen radical levels, which are considered effective factors in the pathogenesis of CRSwNP, can occur due to genetic polymorphism of enzymes in the antioxidant system and genetic polymorphism of antioxidant enzymes in eosinophilic CRSwNP patients might contribute to the pathophysiology.
Assuntos
Catalase/genética , Eosinófilos/patologia , Pólipos Nasais , Procedimentos Cirúrgicos Nasais/métodos , Óxido Nítrico Sintase Tipo II/genética , Rinite , Superóxido Dismutase/genética , Adulto , Antioxidantes/metabolismo , Doença Crônica , Feminino , Humanos , Masculino , Mucosa Nasal/enzimologia , Mucosa Nasal/patologia , Pólipos Nasais/genética , Pólipos Nasais/patologia , Pólipos Nasais/cirurgia , Cirurgia Endoscópica por Orifício Natural/métodos , Estresse Oxidativo/genética , Polimorfismo Genético , Rinite/genética , Rinite/fisiopatologia , Sinusite/genética , Sinusite/fisiopatologiaAssuntos
Enzima de Conversão de Angiotensina 2/biossíntese , Células Epiteliais/efeitos dos fármacos , Interferon gama/farmacologia , Mucosa Nasal/efeitos dos fármacos , Poli I-C/farmacologia , Enzima de Conversão de Angiotensina 2/genética , Células Cultivadas , Indução Enzimática , Células Epiteliais/enzimologia , Feminino , Humanos , Lactente , Masculino , Mucosa Nasal/enzimologiaRESUMO
Naphthalene (NA), a ubiquitous environmental pollutant that can cause pulmonary and nasal toxicity in laboratory animals, requires cytochrome P450 (P450)-mediated metabolic activation to cause toxicity. Our recent study using a Cyp2f2-null mouse showed that CYP2F2 plays an essential role in NA-induced lung toxicity, but not in NA-induced nasal toxicity. The aim of this study was to determine whether mouse CYP2A5, abundantly expressed in nasal olfactory mucosa (OM) and the liver, but less in the lung, plays a major role in the bioactivation and toxicity of NA in the OM. We found, by comparing Cyp2a5-null and wild-type (WT) mice, that the loss of CYP2A5 expression led to substantial decreases in rates of NA metabolic activation by OM microsomes. The loss of CYP2A5 did not cause changes in systemic clearance of NA (at 200 mg/kg, i.p.). However, the Cyp2a5-null mice were much more resistant than were WT mice to NA-induced nasal toxicity (although not lung toxicity), when examined at 24 hours after NA dosing (at 200 mg/kg, i.p.), or to NA-induced depletion of total nonprotein sulfhydryl in the OM (although not in the lung), examined at 2 hours after dosing. Thus, mouse CYP2A5 plays an essential role in the bioactivation and toxicity of NA in the OM, but not in the lung. Our findings further illustrate the tissue-specific nature of the role of individual P450 enzymes in xenobiotic toxicity, and provide the basis for a more reliable assessment of the potential risks of NA nasal toxicity in humans.
Assuntos
Hidrocarboneto de Aril Hidroxilases/metabolismo , Sistema Enzimático do Citocromo P-450/metabolismo , Naftalenos/efeitos adversos , Mucosa Olfatória/metabolismo , Animais , Biotransformação/fisiologia , Citocromo P-450 CYP2A6 , Família 2 do Citocromo P450 , Fígado/enzimologia , Fígado/metabolismo , Masculino , Camundongos , Camundongos Knockout , Microssomos/enzimologia , Microssomos/metabolismo , Mucosa Nasal/enzimologia , Mucosa Nasal/metabolismo , Mucosa Olfatória/enzimologia , Compostos de Sulfidrila/efeitos adversosRESUMO
BACKGROUND: Pseudomonas aeruginosa causes chronic respiratory disease, and the elastase enzyme that it produces increases the permeability of airway epithelial cells owing to the disruption of tight junctions. P. aeruginosa is also implicated in prolonged chronic rhinosinusitis. However, the effects of P. aeruginosa elastase (PE) against the barrier formed by human nasal epithelial cells (HNECs) remain unknown. METHODS: To investigate the mechanisms involved in the disruption of tight junctions by PE in HNECs, primary cultures of HNECs transfected with human telomerase reverse transcriptase (hTERT-HNECs) were used. The hTERT-HNECs were pretreated with inhibitors of various signal transduction pathways, PKC, MAPK, p38MAPK, PI3K, JNK, NF-κB, EGF receptor, proteasome, COX1 and COX2 before treatment with PE. Some cells were pretreated with siRNA and agonist of protease activated receptor-2 (PAR-2) before treatment with PE. Expression and structures of tight junctions were determined by Western blotting, real-time PCR, immunostaining and freeze-fracture. Transepithelial electrical resistance (TER) was examined as the epithelial barrier function. RESULTS: PE treatment transiently disrupted the epithelial barrier and downregulated the transmembrane proteins claudin-1 and -4, occludin, and tricellulin, but not the scaffold PDZ-expression proteins ZO-1 and -2 and adherens junction proteins E-cadherin and ß-catenin. The transient downregulation of tight junction proteins was controlled via distinct signal transduction pathways such as the PKC, MAPK, PI3K, p38 MAPK, JNK, COX-1 and -2, and NF-κB pathways. Furthermore, treatment with PE transiently decreased PAR-2 expression, which also regulated the expression of the tight junction proteins. Treatment with a PAR-2 agonist prevented the downregulation of the tight junction proteins after PE treatment in HNECs. CONCLUSIONS: PE transiently disrupts tight junctions in HNECs and downregulates PAR-2. The transient disruption of tight junctions by PE might occur repeatedly during chronic rhinosinusitis.
Assuntos
Proteínas de Bactérias/fisiologia , Regulação para Baixo/genética , Metaloendopeptidases/fisiologia , Mucosa Nasal/enzimologia , Mucosa Nasal/microbiologia , Elastase Pancreática/fisiologia , Receptor PAR-2/antagonistas & inibidores , Junções Íntimas/enzimologia , Células Cultivadas , Regulação para Baixo/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Mucosa Nasal/metabolismo , Receptor PAR-2/biossíntese , Junções Íntimas/microbiologiaRESUMO
Rhinitis and asthma are the most common respiratory diseases in children. We assessed whether airway inflammation markers were associated with nasal allergies and self-reported symptoms of wheeze and rhinitis in 130 children 6-12 year old in an epidemiological context. Independent of sex and age, the fraction of exhaled nitric oxide (FeNO) and nasal mast cell (MC) activation (tryptase ≥ 5 ng/mL) were positively associated with wheeze, rhinitis and with nasal allergy. Nasal eosinophil cationic protein (ECP) and exhaled breath condensate (EBC) markers (pH, 8-isoprostane, interleukin-1ß) were not associated with symptoms or with nasal allergy. In conclusion, FeNO and nasal tryptase reflect allergic inflammation in the respiratory system.
Assuntos
Mucosa Nasal/enzimologia , Óxido Nítrico/metabolismo , Rinite Alérgica/metabolismo , Triptases/metabolismo , Biomarcadores/metabolismo , Criança , Expiração , Feminino , Humanos , Masculino , Sons RespiratóriosRESUMO
BACKGROUND: Nasal secretion (NS) was investigated as a source of information regarding the mucosal and systemic immune status of cattle challenged by respiratory disease. A method for the collection of substantial volumes (~12 ml) of NS from cattle was developed to establish a reference range of analytes that are present in the NS of healthy cattle. Biochemical profiles of NS from a group of 38 healthy Holstein-Friesian cows revealed high alkaline phosphatase (AP) activity of up to 2392 IU/L. The character and source of the high activity of AP in bovine NS was investigated. RESULTS: Histochemical analysis confirmed the localization of the AP enzyme activity to epithelial cells and serous glands of the nasal respiratory mucosa. Analysis of mRNA levels from nasal mucosa by end point RT-PCR and PCR product sequencing confirmed that the AP was locally produced and is identical at the nucleotide level to the non-specific AP splice variant found in bovine liver, bone and kidney. Analysis by isoelectric focussing confirmed that AP was produced locally at a high level in nasal epithelium demonstrating that AP from nasal secretion and nasal mucosa had similar pI bands, though differing from those of the liver, kidney, bone and intestine, suggesting different post-translational modification (PTM) of AP in these tissues. CONCLUSIONS: A nasal isozyme of AP has been identified that is present at a high activity in NS, resulting from local production and showing distinctive PTM and may be active in NS as an anti-endotoxin mediator.
Assuntos
Fosfatase Alcalina/análise , Bovinos/metabolismo , Mucosa Nasal/metabolismo , Fosfatase Alcalina/genética , Animais , Secreções Corporais/enzimologia , Feminino , Focalização Isoelétrica/veterinária , Mucosa Nasal/enzimologia , Isoformas de Proteínas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterináriaRESUMO
Microbial products play a role in the pathogenesis of allergic diseases; ubiquitin E3 ligase A20 (A20) is an important molecule in regulating inflammation in the body. The present study aims to elucidate the role of A20 in processing the absorbed microbial products in nasal epithelial cells. Human nasal mucosal specimens were collected from patients with or without chronic rhinitis and analyzed by immunohistochemistry. Human nasal epithelial cell line, RPMI2650 cell, was employed to assess the role of A20 in processing the absorbed staphylococcal enterotoxin B (SEB). The RPMI2650 cells absorbed SEB in the culture. The increase in A20 was observed in RPMI2650 cells in parallel to the absorption of SEB. A20 is a critical molecule in the degradation of SEB in the nasal epithelial cells by promoting the tethering of endosomes and lysosomes. A20 plays a critical role in processing of the absorbed SEB in nasal epithelial cells.
Assuntos
Proteínas de Bactérias/metabolismo , Toxinas Bacterianas/metabolismo , Proteínas de Ligação a DNA/metabolismo , Enterotoxinas/metabolismo , Células Epiteliais/enzimologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Mucosa Nasal/enzimologia , Proteínas Nucleares/metabolismo , Proteólise , Staphylococcus aureus/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Adulto , Proteínas de Bactérias/genética , Toxinas Bacterianas/genética , Linhagem Celular , Proteínas de Ligação a DNA/genética , Endossomos/genética , Endossomos/metabolismo , Células Epiteliais/microbiologia , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Lisossomos/genética , Lisossomos/metabolismo , Masculino , Mucosa Nasal/microbiologia , Proteínas Nucleares/genética , Proteína 3 Induzida por Fator de Necrose Tumoral alfa , Ubiquitina-Proteína Ligases/genéticaRESUMO
We have shown that proteinase-activated receptor-2 (PAR(2)) activation in the airways leads to allergic sensitization to concomitantly inhaled Ags, thus implicating PAR(2) in the pathogenesis of asthma. Many aeroallergens with proteinase activity activate PAR(2). To study the role of PAR(2) in allergic sensitization to aeroallergens, we developed a murine model of mucosal sensitization to cockroach proteins. We hypothesized that PAR(2) activation in the airways by natural allergens with serine proteinase activity plays an important role in allergic sensitization. Cockroach extract (CE) was administered to BALB/c mice intranasally on five consecutive days (sensitization phase) and a week later for four more days (challenge phase). Airway hyperresponsiveness (AHR) and allergic airway inflammation were assessed after the last challenge. To study the role of PAR(2), mice were exposed intranasally to a receptor-blocking anti-PAR(2) Ab before each administration of CE during the sensitization phase. Mucosal exposure to CE induced eosinophilic airway inflammation, AHR, and cockroach-specific IgG1. Heat-inactivated or soybean trypsin inhibitor-treated CE failed to induce these effects, indicating that proteinase activity plays an important role. The use of an anti-PAR(2) blocking Ab during the sensitization phase completely inhibited airway inflammation and also decreased AHR and the production of cockroach-specific IgG1. PAR(2) activation by CE acts as an adjuvant for allergic sensitization even in the absence of functional TLR4. We conclude that CE induces PAR(2)-dependent allergic airway sensitization in a mouse model of allergic airway inflammation. PAR(2) activation may be a general mechanism used by aeroallergens to induce allergic sensitization.
Assuntos
Alérgenos/imunologia , Blattellidae/imunologia , Receptor PAR-2/metabolismo , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/metabolismo , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/fisiologia , Administração Intranasal , Alérgenos/administração & dosagem , Animais , Blattellidae/enzimologia , Hiper-Reatividade Brônquica/enzimologia , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/metabolismo , Linhagem Celular Transformada , Modelos Animais de Doenças , Ativação Enzimática/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mucosa Nasal/enzimologia , Mucosa Nasal/imunologia , Mucosa Nasal/metabolismo , Ratos , Receptor PAR-2/deficiência , Receptor PAR-2/imunologia , Hipersensibilidade Respiratória/enzimologiaRESUMO
OBJECTIVES: We investigated the role of inducible nitric oxide synthase (iNOS) in the pathogenesis of sinonasal polyps. METHODS: Adult patients (21 men, 3 women) with nasal polyposis underwent functional endoscopic sinus surgery. Nine adults without polyps (6 men) who underwent septoplasty and/or rhinoplasty served as controls. Polyp specimens came from three regions: the maxillary sinus (10), ethmoid sinus (14), and nasal cavity (10). Control group samples (9) came from the inferior turbinate. Specimens were evaluated in eight mucosal layers for count and distribution of inflammatory cells and iNOS expression. An iNOS positivity index (PI) was determined for the epithelium (E), subepithelial layer of the lamina propria (SE), and deep paraglandular layer of the mucosa (D). RESULTS: Polymorphonuclear cell (PMNC) % values of the ethmoid and maxillary sinus and overall ethmoid sinus PI were significantly higher in the polyp group. Patients with longer polyp duration, D-perivascular (D-pv), and a higher Brinkmann index had decreased ethmoid sinus D PIs. However, in older patients and patients with longer polyp duration, perivascular PIs increased in maxillary sinus SE and D, respectively. Furthermore, as PMNC % and iNOS-PMNC PI increased, SE_glandular and epithelial_apical iNOS values decreased. In the ethmoid and maxillary sinuses, iNOS_D_. endothelial values increased but decreased in the nasal cavity. CONCLUSIONS: iNOS may play a role in sinonasal polyp pathogenesis, especially in mucosal SE and D layers. Increased vascular permeability, stromal edema, inflammatory cell migration into the stroma of the mucosa, and increased mucosal gland secretion may result in polyp formation.
Assuntos
Mucosa Nasal/enzimologia , Pólipos Nasais/enzimologia , Óxido Nítrico Sintase Tipo II/metabolismo , Doenças dos Seios Paranasais/enzimologia , Adulto , Estudos de Casos e Controles , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Pólipos Nasais/etiologia , Neutrófilos/citologia , Doenças dos Seios Paranasais/etiologia , Pólipos/enzimologia , Pólipos/etiologiaRESUMO
Matrix metalloproteinases (MMPs) play an important role in respiratory inflammatory diseases, such as asthma and chronic obstructive pulmonary disease. It was hypothesized that MMP-8 and MMP-9 may function as biological markers to assess disease severity in viral lower respiratory tract infections in children. MMP-8 and MMP-9 mRNA expression levels in peripheral blood mononuclear cells (PBMCs) and granulocytes obtained in both the acute and recovery phase from 153 children with mild, moderate, and severe viral lower respiratory tract infections were determined using real-time PCR. In addition, MMP-8 and MMP-9 concentrations in blood and nasopharyngeal specimens were determined during acute mild, moderate, and severe infection, and after recovery using ELISA. Furthermore, PBMCs and neutrophils obtained from healthy volunteers were stimulated with RSV, LPS (TLR4 agonist), and Pam3Cys (TLR2 agonist) in vitro. Disease severity of viral lower respiratory tract infections in children is associated with increased expression levels of the MMP-8 and MMP-9 genes in both PBMCs and granulocytes. On the contrary, in vitro experiments showed that MMP-8 and MMP-9 mRNA and protein expression in PBMCs and granulocytes is not induced by stimulation with RSV, the most frequent detected virus in young children with viral lower respiratory tract infections. These data indicate that expression levels of the MMP-8 and MMP-9 genes in both PBMCs and neutrophils are associated with viral lower respiratory tract infections disease severity. These observations justify future validation in independent prospective study cohorts of the usefulness of MMP-8 and MMP-9 as potential markers for disease severity in viral respiratory infections.
Assuntos
Metaloproteinase 8 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Infecções por Vírus Respiratório Sincicial/enzimologia , Vírus Sinciciais Respiratórios , Infecções Respiratórias/enzimologia , Feminino , Expressão Gênica , Granulócitos/enzimologia , Células HeLa , Humanos , Lactente , Leucócitos Mononucleares/enzimologia , Masculino , Metaloproteinase 8 da Matriz/sangue , Metaloproteinase 8 da Matriz/genética , Metaloproteinase 9 da Matriz/sangue , Metaloproteinase 9 da Matriz/genética , Mucosa Nasal/enzimologia , Mucosa Nasal/virologia , Neutrófilos/enzimologia , Neutrófilos/virologia , Faringe/enzimologia , Faringe/virologia , Estudos Prospectivos , Infecções por Vírus Respiratório Sincicial/patologia , Infecções por Vírus Respiratório Sincicial/virologia , Infecções Respiratórias/patologia , Infecções Respiratórias/virologia , Índice de Gravidade de Doença , Estatísticas não ParamétricasRESUMO
CYP2A13, CYP2B6, and CYP2F1, which are encoded by neighboring cytochrome P450 genes on human chromosome 19, are active in the metabolic activation of many drugs, respiratory toxicants, and chemical carcinogens. To facilitate studies on the regulation and function of these human genes, we have generated a CYP2A13/2B6/2F1-transgenic (TG) mouse model (all *1 alleles). Homozygous transgenic mice are normal with respect to gross morphological features, development, and fertility. The tissue distribution of transgenic mRNA expression agreed well with the known respiratory tract-selective expression of CYP2A13 and CYP2F1 and hepatic expression of CYP2B6 in humans. CYP2A13 protein was detected through immunoblot analyses in the nasal mucosa (NM) (â¼100 pmol/mg of microsomal protein; similar to the level of mouse CYP2A5) and the lung (â¼0.2 pmol/mg of microsomal protein) but not in the liver of the TG mice. CYP2F1 protein, which could not be separated from mouse CYP2F2 in immunoblot analyses, was readily detected in the NM and lung but not the liver of TG/Cyp2f2-null mice, at levels 10- and 40-fold, respectively, lower than that of mouse CYP2F2 in the TG mice. CYP2B6 protein was detected in the liver (â¼0.2 pmol/mg of microsomal protein) but not the NM or lung (with a detection limit of 0.04 pmol/mg of microsomal protein) of the TG mice. At least one transgenic protein (CYP2A13) seems to be active, because the NM of the TG mice had greater in vitro and in vivo activities in bioactivation of a CYP2A13 substrate, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (a lung carcinogen), than did the NM of wild-type mice.
Assuntos
Hidrocarboneto de Aril Hidroxilases/biossíntese , Sistema Enzimático do Citocromo P-450/biossíntese , Modelos Animais , Família Multigênica/genética , Oxirredutases N-Desmetilantes/biossíntese , Adolescente , Animais , Hidrocarboneto de Aril Hidroxilases/química , Hidrocarboneto de Aril Hidroxilases/genética , Citocromo P-450 CYP2B6 , Sistema Enzimático do Citocromo P-450/deficiência , Sistema Enzimático do Citocromo P-450/genética , Família 2 do Citocromo P450 , Feminino , Humanos , Fígado/enzimologia , Pulmão/enzimologia , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Mucosa Nasal/enzimologia , Oxirredutases N-Desmetilantes/química , Oxirredutases N-Desmetilantes/genética , Distribuição Tecidual/genéticaRESUMO
BACKGROUND: Nitric oxide (NO) has various roles in airway physiology and pathophysiology. Monitoring exhaled NO levels is increasingly common to measure airways inflammation and inhaled NO studied for its therapeutic value in premature infants and adult respiratory distress syndrome. NO is produced by 3 isoforms of NO synthase (NOS1, 2, 3), and each can play distinct and perhaps overlapping roles in the airways. However, the distribution, regulation, and functions of NOS in various cells in the upper airways, particularly in leukocytes, are incompletely understood. OBJECTIVE: To characterize the expression of NOS isoforms in leukocytes in normal middle turbinate tissues (MT) and in inflammatory nasal tissue (nasal polyps, NP). METHODS: Normal MT tissue was collected from surgical specimens that were to be discarded. The NP samples were from surgical tissue archives of 15 patients with chronic rhinosinusitis. Isoforms of NOS in cells were identified by double immunostaining using NOS isoform-specific and leukocyte-specific (mast cell, eosinophil, macrophage, neutrophil, or T cell) antibodies. RESULTS: The proportion of total cells below the epithelium that were positive for each isoform of NOS was higher in NP than in MT. Each isoform of NOS was found in all leukocyte populations studied, and there were significant differences in the percentage of leukocytes expressing NOS isoforms between MT and NP. CONCLUSION: All isoforms of NOS are expressed in leukocytes in MT and NP, and their expression varies among leukocyte types. Our data provide a basis to investigate the regulation, cell distribution, and distinct functions of NOS isoforms in normal and inflamed nasal tissues.
Assuntos
Leucócitos/enzimologia , Pólipos Nasais/enzimologia , Óxido Nítrico Sintase Tipo III/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Óxido Nítrico Sintase Tipo I/metabolismo , Adulto , Eosinófilos/enzimologia , Eosinófilos/imunologia , Feminino , Humanos , Inflamação/imunologia , Leucócitos/imunologia , Macrófagos/enzimologia , Macrófagos/imunologia , Masculino , Mastócitos/enzimologia , Mastócitos/imunologia , Pessoa de Meia-Idade , Mucosa Nasal/enzimologia , Mucosa Nasal/imunologia , Mucosa Nasal/patologia , Pólipos Nasais/imunologia , Pólipos Nasais/patologia , Óxido Nítrico/biossíntese , Conchas Nasais/enzimologiaRESUMO
In this study, we evaluated effect of glycyrrhizin on immunity function in allergic rhinitis (AR) mice. The AR mice model were induced by dripping ovalbumin in physiological saline (2 mg mL⻹, 10 µL) into the bilateral nasal cavities using a micropipette. After the AR model was induced, mice were randomly divided into six groups: the normal control, model, lycopene 20 mg kg⻹ (as positive control drug) group, and glycyrrhizin 10, 20, 30 mg kg⻹ groups. After the sensitization day 14, lycopene (20 mg/kg BW) and glycyrrhizin (10, 20 and 30 mg/kg BW) were given orally for 20 days once a day. Mice in the normal control and model groups were given saline orally once a day for 20 days. Results showed that glycyrrhizin treatment could dose-dependently significantly reduce blood immunoglobulin E (IgE), interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-6 (IL-6), nitrous oxide (NO), tumor necrosis factor-alpha (TNF-α) levels and nitrous oxide synthase (NOS) activity and enhance blood immunoglobulin A (IgA), immunoglobulin G (IgG), immunoglobulin M (IgM), interleukin-2 (IL-2) and interleukin-12 (IL-12) levels in AR mice. Furthermore, glycyrrhizin treatment could dose-dependently significantly enhance acetylcholinesterase (AchE) activity and reduce substance P (SP) level in peripheral blood and nasal mucosa of AR mice. We conclude that glycyrrhizin can improve immunity function in AR mice, suggesting a potential drug for the prevention and therapy of AR.
Assuntos
Anti-Inflamatórios/farmacologia , Ácido Glicirrízico/farmacologia , Rinite Alérgica Perene/imunologia , Acetilcolinesterase/metabolismo , Animais , Anti-Inflamatórios/uso terapêutico , Ácido Glicirrízico/uso terapêutico , Imunoglobulinas/sangue , Interleucinas/sangue , Masculino , Camundongos , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/enzimologia , Mucosa Nasal/metabolismo , Óxido Nítrico/sangue , Óxido Nítrico Sintase/sangue , Ovalbumina , Distribuição Aleatória , Ratos , Rinite Alérgica Perene/sangue , Rinite Alérgica Perene/induzido quimicamente , Rinite Alérgica Perene/tratamento farmacológico , Substância P/sangue , Substância P/metabolismo , Fator de Necrose Tumoral alfa/sangueRESUMO
Odor information is decoded by a combination of odorant receptors, and thus transformed into discrete spatial patterns of olfactory glomerular activity. It has been found, however, that for some odorants, there are differences between the ligand specificity of an odorant receptor in vitro and its corresponding glomerulus in vivo. These observations led us to hypothesize that there exist prereceptor events that affect the local concentration of a given odorant in the nasal mucus, thus causing the apparent specificity differences. Here we show that odorants with functional groups such as aldehydes and esters are targets of metabolic enzymes secreted in the mouse mucus, resulting in their conversion to the corresponding acids and alcohols. The glomerular activation patterns elicited by an enzyme-targeted odorant in the olfactory bulb was different in the presence of an enzyme inhibitor in the mucosa, suggesting that the enzymatic conversion occurs fast enough to affect recognition of the odorant at the levels of olfactory sensory neurons. Importantly, olfactory discrimination tests revealed that mice behaviorally trained to associate an enzyme-targeted odorant to sugar rewards could not discriminate the odorant after treatment with the enzyme inhibitor. These results reveal that the enzymatic conversion of odorants in the nasal mucus appears be fast enough to affect olfactory perception, which sheds light on the previously unappreciated role of nasal mucosal enzymes in odor sensation.
Assuntos
Mucosa Nasal/metabolismo , Odorantes , Percepção Olfatória/fisiologia , Receptores Odorantes/metabolismo , Animais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Mucosa Nasal/enzimologia , Odorantes/análiseRESUMO
To identify novel factors or mechanisms that are important for the resistance of tissues to chemical toxicity, we have determined the mechanisms underlying the previously observed increases in resistance to acetaminophen (APAP) toxicity in the lateral nasal gland (LNG) of the male Cyp2g1-null/Cyp2a5-low mouse. Initial studies established that Cyp2a5-null mice, but not a newly generated strain of Cyp2g1-null mice, were resistant to APAP toxicity in the LNG; therefore, subsequent studies were focused on the Cyp2a5-null mice. Compared with the wild-type (WT) male mouse, the Cyp2a5-null male mouse had intact capability to metabolize APAP to reactive intermediates in the LNG, as well as unaltered circulating levels of APAP, APAP-GSH, APAP-glucuronide, and APAP-sulfate. However, it displayed reduced tissue levels of APAP and APAP-GSH and increased tissue levels of testosterone and salivary androgen-binding protein (ABP) in the LNG. Furthermore, we found that ABP was able to compete with GSH and cellular proteins for adduction with reactive metabolites of APAP in vitro. The amounts of APAP-ABP adducts formed in vivo were greater, whereas the amounts of APAP adducts formed with other cellular proteins were substantially lower, in the LNG of APAP-treated male Cyp2a5-null mice compared with the LNG of APAP-treated male WT mice. We propose that through its critical role in testosterone metabolism, CYP2A5 regulates 1) the bioavailability of APAP and APAP-GSH (presumably through modulation of the rates of xenobiotic excretion from the LNG) and 2) the expression of ABP, which can quench reactive APAP metabolites and thereby spare critical cellular proteins from inactivation.
Assuntos
Acetaminofen/toxicidade , Proteína de Ligação a Androgênios/biossíntese , Hidrocarboneto de Aril Hidroxilases/fisiologia , Mucosa Nasal/metabolismo , Proteínas e Peptídeos Salivares/biossíntese , Testosterona/fisiologia , Acetaminofen/farmacocinética , Animais , Disponibilidade Biológica , Citocromo P-450 CYP2A6 , Família 2 do Citocromo P450 , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/fisiologia , Homeostase/efeitos dos fármacos , Homeostase/fisiologia , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/enzimologiaRESUMO
Genotoxicity experiments with exfoliated nasal mucosa cells are a promising minimally invasive approach for the detection of DNA-damaging compounds in ambient air. Results of single cell gel electrophoresis (SCGE) assays with individual cells and organ cultures from bioptic material show that DNA damage caused by compounds such as nitrosamines, polycyclic aromatic hydrocarbons and pesticides can be detected. Biochemical studies indicate that enzymes involved in the metabolism of environmental mutagens are represented in nasal cells. Several protocols for experiments with nasal cells have been developed and it was shown that formaldehyde, metals, styrene and crystalline silica induce DNA damage in SCGE and/or in micronucleus studies; furthermore, it was also found that polluted urban air causes DNA instability in nasal epithelial cells. Comparisons of these data with results obtained in lymphocytes and buccal cells indicate that nasal cells are in general equally sensitive. Broad variations in the baseline levels, differences of results obtained in various studies as well as the lack of information concerning the impact of confounding factors on the outcome of experiments with these cells indicate the need for further standardisation of the experimental protocols.
Assuntos
Dano ao DNA , Mutagênicos/toxicidade , Mucosa Nasal/efeitos dos fármacos , Animais , Ensaio Cometa , Humanos , Inativação Metabólica , Testes para Micronúcleos , Mucosa Nasal/enzimologia , Mucosa Nasal/ultraestrutura , Nariz/citologia , Nariz/enzimologia , RoedoresRESUMO
BACKGROUND: Matrix metalloproteinases (MMPs) are key enzymes responsible for extracellular matrix degradation contributing to the progressive histological changes seen in lower airway disease, including asthma. MMP-9 and TIMP-1 have also shown some role in the pathogenesis of chronic rhinosinusitis (CRS) and nasal polyposis (NP). OBJECTIVE: We aim to determine variability in expression of MMP-9 and its inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1), in sinus tissue from distinct patient populations presenting with nasal polyposis. METHODS: The expression of MMP-9 and TIMP-1 was investigated in nasal polyp tissue from 6 aspirin-sensitive (AS) and 6 aspirin-tolerant (AT) patients undergoing endoscopic sinus surgery for chronic rhinosinusitis with nasal polyposis (CRSwNP). Sinus mucosa from 6 patients with chronic rhinosinusitis without nasal polyposis (CRSsNP) was used as control. The MMP-9 and TIMP-1 expression was measured using immunofluorescence technique and graded using manual and computerized methods. RESULTS: Expression of TIMP-1 was significantly reduced in the AS group when compared with both the AT and CRSsNP (control) groups (P < .001). The MMP-9/TIMP-1 ratio was significantly increased in the AS group when compared with other patient groups (P < .001). The MMP- 9 expression was similar between study and control groups. CONCLUSION: These results support the importance of MMP-9 and TIMP-1 expression in nasal polyp formation. The decreased expression of TIMP-1 in AS patients may promote the effects of MMP-9 expression and thus contribute to tissue remodeling and inflammatory changes. This finding may lead to further understanding of disease severity and resistance to treatment in this group of patients, as well as the pathogenesis of nasal polyps.
Assuntos
Anti-Inflamatórios não Esteroides/administração & dosagem , Aspirina/administração & dosagem , Metaloproteinase 9 da Matriz/biossíntese , Pólipos Nasais/metabolismo , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Adulto , Anti-Inflamatórios não Esteroides/efeitos adversos , Aspirina/efeitos adversos , Biópsia , Feminino , Histocitoquímica , Humanos , Masculino , Microscopia de Fluorescência , Pessoa de Meia-Idade , Mucosa Nasal/efeitos dos fármacos , Mucosa Nasal/enzimologia , Mucosa Nasal/cirurgia , Pólipos Nasais/enzimologia , Pólipos Nasais/cirurgia , Estudos Retrospectivos , Adulto JovemRESUMO
Dexamethasone cipecilate (DX-CP, 9-fluoro-11ß,17,21-trihydroxy-16α-methylpregna-1,4-diene-3,20-dione 21-cyclohexanecarboxylate 17-cyclopropanecarboxylate) is a novel synthetic corticosteroid used to treat allergic rhinitis. The pharmacological effect of DX-CP is considered to be mainly due to its active de-esterified metabolite (DX-17-CPC). To investigate the in vitro metabolism of DX-CP in human liver, DX-CP was incubated with human liver microsomes and S9. In addition, a metabolism study of DX-CP with human nasal mucosa was carried out in order to elucidate whether DX-17-CPC is formed in nasal mucosa, the site of action of DX-CP. DX-17-CPC was the major metabolite in both liver microsomes and S9. Two new epoxide metabolites, UK1 and UK2, were detected in liver S9, while only UK1 was detected in liver microsomes. This suggests that cytosol enzymes are responsible for the formation of UK2. In human nasal mucosa, DX-CP was mainly transformed into DX-17-CPC. By using recombinant human carboxylesterases (CESs), the reaction was shown to be catalyzed by CES2. These results provide the evidence that the active metabolite DX-17-CPC is the main contributor to the pharmacological action after the intranasal administration of DX-CP to humans.