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1.
Mol Med Rep ; 27(2)2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36601769

RESUMO

The presence of allergic rhinitis (AR) is an increased risk factor for the occurrence of bronchial asthma (BA). Nerve growth factor (NGF), in addition to its key role in the development and differentiation of neurons, may also be an important inflammatory factor in AR and BA. However, the pathogenesis of the progression of AR to BA remains to be elucidated. The present study aimed to investigate the ability of NGF to mediate nasobronchial interactions and explore possible underlying molecular mechanisms. In the present study, an AR mouse model was established and histology of nasal mucosa tissue injury was determined. The level of phenylethanolamine N­methyl transferase in adrenal medulla was determined by immunofluorescence. Primary adrenal medullary chromaffin cells (AMCCs) were isolated and cultured from the adrenal medulla of mice. The expression levels of synaptophysin (SYP), STAT1, JAK1, p38 and ERK in NGF­treated and untreated AMCCs were detected by reverse­transcription­quantitative PCR and western blotting. The epinephrine (EPI) and norepinephrine (NE) concentrations were measured by ELISA. It was found that the expression of SYP in AMCCs was enhanced in the presence of NGF, whereas, the concentration of EPI decreased significantly under the same conditions. Furthermore, NGF mediated the phenotypic and functional changes of AMCCs, resulting in decreased EPI secretion via JAK1/STAT1, p38 and ERK signaling. In conclusion, these findings could provide novel evidence for the role of NGF in regulating neuroendocrine mechanisms.


Assuntos
Asma , Células Cromafins , Rinite Alérgica , Ratos , Animais , Camundongos , Fator de Crescimento Neural/genética , Fator de Crescimento Neural/metabolismo , Ratos Sprague-Dawley , Epinefrina/farmacologia , Asma/metabolismo , Rinite Alérgica/metabolismo , Células Cromafins/metabolismo , Fenótipo
2.
BMC Pulm Med ; 23(1): 45, 2023 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-36717790

RESUMO

BACKGROUND: Micro RNA (miRNA) plays important roles in macrophage polarization. However, the manner in which miRNA regulate macrophage polarization in response to dermatophagoides farinae protein 1(Der f1)-induced asthma has not been defined. This study aims to explore the role of miRNAs in regulating macrophages in asthma. METHODS: The microRNAs which may regulate asthma were selectd by Microarrays. The function of miR-125b-5p in macrophage and Der f1-induced asthma were detected in vivo experiment. The long non coding RNA (lncRNA) AK089514/miR-125b-5p/TRAF6 axis was predicted by bioinformatics and confirmed by dual luciferase reporter assay. RESULTS: In this study, we found that miR-125b-5p is highly expressed in M2 macrophages and bronchoalveolar lavage fluid (BALF) cells with Der f1-induced asthma. In response to the challenge of Der f1, miR-125b-5p KD attenuated allergic airway inflammation of mice by preventing M2 macrophages polarization. Mechanistic studies indicated that lncRNA AK089514 functioned as a competing endogenous RNA for miR-125b-5p, thereby leading to the depression of its endogenous target TNF receptor associated factor 6 (TRAF6). CONCLUSIONS: miR-125b-5p is significantly over-expressed in asthma, and AK089514-miR-125b-5p-TRAF6 axis play critical role in asthma by modulating macrophage polarization. Our findings may provide a potential new target for potential therapeutic and diagnostic target in asthma.


Assuntos
Asma , Macrófagos , MicroRNAs , RNA Longo não Codificante , Fator 6 Associado a Receptor de TNF , Animais , Camundongos , Asma/genética , Asma/metabolismo , Inflamação/metabolismo , Macrófagos/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Fator 6 Associado a Receptor de TNF/genética , Fator 6 Associado a Receptor de TNF/metabolismo
3.
Biol Sex Differ ; 14(1): 2, 2023 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-36609358

RESUMO

RATIONALE: Asthma is a chronic airway condition that occurs more often in women than men during reproductive years. Population studies have collectively shown that long-term use of oral contraceptives decreased the onset of asthma in women of reproductive age. In the current study, we hypothesized that steady-state levels of estrogen would reduce airway inflammation and airway hyperresponsiveness to methacholine challenge. METHODS: Ovariectomized BALB/c mice (Ovx) were implanted with subcutaneous hormone pellets (estrogen, OVX-E2) that deliver consistent levels of estrogen [68 ± 2 pg/mL], or placebo pellets (OVX-Placebo), followed by ovalbumin sensitization and challenge. In conjunction with methacholine challenge, immune phenotyping was performed to correlate inflammatory proteins and immune populations with better or worse pulmonary outcomes measured by invasive pulmonary mechanics techniques. RESULTS: Histologic analysis showed an increase in total cell infiltration and mucus staining around the airways leading to an increased inflammatory score in ovarectomized (OVX) animals with steady-state estrogen pellets (OVX-E2-OVA) as compared to other groups including female-sham operated (F-INTACT-OVA) and OVX implanted with a placebo pellet (OVX-Pl-OVA). Airway resistance (Rrs) and lung elastance (Ers) were increased in OVX-E2-OVA in comparison to F-INTACT-OVA following aerosolized intratracheal methacholine challenges. Immune phenotyping revealed that steady-state estrogen reduced CD3+ T cells, CD19+ B cells, ILC2 and eosinophils in the BAL across all experiments. While these commonly described allergic cells were reduced in the BAL, or airways, we found no changes in neutrophils, CD3+ T cells or CD19+ B cells in the remaining lung tissue. Similarly, inflammatory cytokines (IL-5 and IL-13) were also decreased in OVX-E2-OVA-treated animals in comparison to Female-INTACT-OVA mice in the BAL, but in the lung tissue IL-5, IL-13 and IL-33 were comparable in OVX-E2-OVA and F-INTACT OVA mice. ILC2 were sorted from the lungs and stimulated with exogenous IL-33. These ILC2 had reduced cytokine and chemokine expression when they were isolated from OVX-E2-OVA animals, indicating that steady-state estrogen suppresses IL-33-mediated activation of ILC2. CONCLUSIONS: Therapeutically targeting estrogen receptors may have a limiting effect on eosinophils, ILC2 and potentially other immune populations that may improve asthma symptoms in those females that experience perimenstrual worsening of asthma, with the caveat, that long-term use of estrogens or hormone receptor modulators may be detrimental to the lung microenvironment over time.


Assuntos
Asma , Interleucina-33 , Feminino , Animais , Camundongos , Interleucina-33/uso terapêutico , Estradiol/farmacologia , Estradiol/uso terapêutico , Imunidade Inata , Interleucina-13/uso terapêutico , Cloreto de Metacolina/farmacologia , Cloreto de Metacolina/uso terapêutico , Alérgenos/uso terapêutico , Resistência das Vias Respiratórias , Interleucina-5/uso terapêutico , Líquido da Lavagem Broncoalveolar , Linfócitos/metabolismo , Linfócitos/patologia , Pulmão/metabolismo , Asma/tratamento farmacológico , Asma/metabolismo , Citocinas , Estrogênios/uso terapêutico
4.
Nat Commun ; 14(1): 47, 2023 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-36599824

RESUMO

Obesity increases asthma prevalence and severity. However, the underlying mechanisms are poorly understood, and consequently, therapeutic options for asthma patients with obesity remain limited. Here we report that cholecystokinin-a metabolic hormone best known for its role in signaling satiation and fat metabolism-is increased in the lungs of obese mice and that pharmacological blockade of cholecystokinin A receptor signaling reduces obesity-associated airway hyperresponsiveness. Activation of cholecystokinin A receptor by the hormone induces contraction of airway smooth muscle cells. In vivo, cholecystokinin level is elevated in the lungs of both genetically and diet-induced obese mice. Importantly, intranasal administration of cholecystokinin A receptor antagonists (proglumide and devazepide) suppresses the airway hyperresponsiveness in the obese mice. Together, our results reveal an unexpected role for cholecystokinin in the lung and support the repurposing of cholecystokinin A receptor antagonists as a potential therapy for asthma patients with obesity.


Assuntos
Asma , Hipersensibilidade Respiratória , Animais , Camundongos , Asma/tratamento farmacológico , Asma/metabolismo , Colecistocinina/metabolismo , Pulmão/metabolismo , Camundongos Obesos , Obesidade/complicações , Obesidade/metabolismo , Receptor de Colecistocinina A/genética , Receptor de Colecistocinina A/metabolismo , Hipersensibilidade Respiratória/tratamento farmacológico , Hipersensibilidade Respiratória/metabolismo
5.
J Exp Clin Cancer Res ; 42(1): 26, 2023 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-36670473

RESUMO

BACKGROUND: Individuals with certain chronic inflammatory lung diseases have a higher risk of developing lung cancer (LC). However, the underlying mechanisms remain largely unknown. Here, we hypothesized that chronic exposure to house dust mites (HDM), a common indoor aeroallergen associated with the development of asthma, accelerates LC development through the induction of chronic lung inflammation (CLI).  METHODS: The effects of HDM and heat-inactivated HDM (HI-HDM) extracts were evaluated in two preclinical mouse models of LC (a chemically-induced model using the carcinogen urethane and a genetically-driven model with oncogenic KrasG12D activation in lung epithelial cells) and on murine macrophages in vitro. Pharmacological blockade or genetic deletion of the Nod-like receptor family pyrin domain-containing protein 3 (NLRP3) inflammasome, caspase-1, interleukin-1ß (IL-1ß), and C-C motif chemokine ligand 2 (CCL2) or treatment with an inhaled corticosteroid (ICS) was used to uncover the pro-tumorigenic effect of HDM.  RESULTS: Chronic intranasal (i.n) instillation of HDM accelerated LC development in the two mouse models. Mechanistically, HDM caused a particular subtype of CLI, in which the NLRP3/IL-1ß signaling pathway is chronically activated in macrophages, and made the lung microenvironment conducive to tumor development. The tumor-promoting effect of HDM was significantly decreased by heat treatment of the HDM extract and was inhibited by NLRP3, IL-1ß, and CCL2 neutralization, or ICS treatment. CONCLUSIONS: Collectively, these data indicate that long-term exposure to HDM can accelerate lung tumorigenesis in susceptible hosts (e.g., mice and potentially humans exposed to lung carcinogens or genetically predisposed to develop LC).


Assuntos
Asma , Neoplasias Pulmonares , Humanos , Animais , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Pyroglyphidae , Pulmão/patologia , Asma/metabolismo , Asma/patologia , Neoplasias Pulmonares/induzido quimicamente , Neoplasias Pulmonares/metabolismo , Modelos Animais de Doenças , Microambiente Tumoral
6.
Mol Med Rep ; 27(1)2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36321783

RESUMO

Allergic asthma is a chronic inflammatory disease in which oxidative stress serves a pivotal role. In clinical practice, dexmedetomidine (DEX), an α­2­adrenergic receptor agonist, is used as a sedative. DEX exhibits antioxidative and organ­protective properties. In a murine model of asthma, DEX has a therapeutic effect via the toll like receptor 4/NF­ÐºB signaling pathway; however, whether DEX can exert an antioxidative effect on asthma has yet to be elucidated. In the present study, a T helper (Th)2­dominant murine asthma model was established. DEX treatment significantly reduced eosinophilic airway inflammation, mucus overproduction and airway hyperresponsiveness, as well as the concentrations of Th2 cytokines. The lung tissues of mice with asthma were characterized by redox imbalance (increased oxidative stress and impaired antioxidant capacity). DEX treatment alleviated this imbalance by decreasing the levels of malondialdehyde and reactive oxygen species, and increasing the levels of glutathione. Furthermore, the nuclear factor erythroid 2­related factor 2 (Nrf2) signaling pathway was inhibited in the lung tissues of asthmatic mice; these effects were noted in its downstream genes, heme oxygenase 1 and glutathione peroxidase 4. In mice with asthma, DEX treatment induced the expression of these antioxidant genes and the activation of Nrf2, whereas ML385 (an inhibitor of Nrf2) partially abrogated the antioxidative and therapeutic effects of DEX. To the best of our knowledge, the present study is the first to demonstrate the protective effect of DEX on Th2­dominant asthma through the activation of the Nrf2 signaling pathway. The results suggested that the antioxidative properties of DEX could be beneficial in clinical application of DEX for the relief of asthmatic symptoms.


Assuntos
Asma , Dexmedetomidina , Animais , Camundongos , Antioxidantes/farmacologia , Asma/metabolismo , Dexmedetomidina/farmacologia , Inflamação/metabolismo , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Transdução de Sinais
7.
Pulm Pharmacol Ther ; 78: 102183, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36481301

RESUMO

INTRODUCTION: In most asthma patients, symptoms are controlled by treatment with glucocorticoid, but long-term or high-dose use can produce adverse effects. Therefore, it is crucial to find new therapeutic strategies. ß-sitosterol could suppress type Ⅱ inflammation in ovalbumin (OVA)-induced mice, but its mechanisms have remained unclear. METHODS: A binding activity of ß-sitosterol with glucocorticoid receptor (GR) was analyzed by molecular docking. Human bronchial epithelial cells (BEAS-2B) and human bronchial smooth muscle cells (HBSMC) were treated with different concentrations (0, 1, 5, 10, 20, and 50 µg/mL) of ß-sitosterol for suitable concentration selection. In transforming growth factor (TGF)-ß1 treated BEAS-2B and HBSMC, cells were treated with 20 µg/mL ß-sitosterol or dexamethasone (Dex) to analyze its possible mechanism. In OVA-induced mice, 2.5 mg/kg ß-sitosterol or Dex administration was performed to analyze the therapeutic mechanism of ß-sitosterol. A GR antagonist RU486 was used to confirm the mechanism of ß-sitosterol in the treatment of asthma. RESULTS: A good binding of ß-sitosterol to GR (score = -8.2 kcal/mol) was found, and the GR expression was upregulated with ß-sitosterol dose increase in BEAS-2B and HBSMC. Interleukin (IL)-25 and IL-33 secretion was significantly decreased by ß-sitosterol in the TGF-ß1-induced BEAS-2B, and the levels of collagen 1A and α-smooth muscle actin (SMA) were reduced in the TGF-ß1-induced HBSMC. In the OVA-challenged mice, ß-sitosterol treatment improved airway inflammation and remodeling through suppressing type Ⅱ immune response and collagen deposition. The therapeutic effects of ß-sitosterol were similar to Dex treatment in vitro and in vivo. RU486 treatment clearly hampered the therapeutic effects of ß-sitosterol in the TGF-ß1-induced cells and OVA-induced mice. CONCLUSION: This study identified that ß-sitosterol binds GR to perform its functions in asthma treatment. ß-sitosterol represent a potential therapeutic drug for allergic asthma.


Assuntos
Asma , Receptores de Glucocorticoides , Sitosteroides , Animais , Humanos , Camundongos , Remodelação das Vias Aéreas , Asma/tratamento farmacológico , Asma/metabolismo , Colágeno/metabolismo , Modelos Animais de Doenças , Inflamação/tratamento farmacológico , Pulmão , Camundongos Endogâmicos BALB C , Mifepristona/farmacologia , Mifepristona/uso terapêutico , Simulação de Acoplamento Molecular , Ovalbumina , Receptores de Glucocorticoides/metabolismo , Fator de Crescimento Transformador beta1/farmacologia , Sitosteroides/farmacologia
8.
Cytokine ; 162: 156091, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36481478

RESUMO

RATIONALE: Type 2 (T2) asthma is characterized by airflow limitations and elevated levels of blood and sputum eosinophils, fractional exhaled nitric oxide, IgE, and periostin. While eosinophils are associated with exacerbations, the contribution of eosinophils to lung inflammation, remodeling and function remains largely hypothetical. OBJECTIVES: To determine the effect of T2 cytokines IL-4, IL-13 and IL-5 on eosinophil biology and compare the impact of depleting just eosinophils versus inhibiting all aspects of T2 inflammation on airway inflammation. METHODS: Human eosinophils or endothelial cells stimulated with IL-4, IL-13 or IL-5 were assessed for gene changes or chemokine release.Mice exposed to house dust mite extract received anti-IL-4Rα (dupilumab), anti-IL-5 or control antibodies and were assessed for changes in lung histological and inflammatory endpoints. MEASUREMENTS AND MAIN RESULTS: IL-4 or IL-13 stimulation of human eosinophils and endothelial cells induced gene expression changes related to granulocyte migration; whereas, IL-5 induced changes reflecting granulocyte differentiation.In a mouse model, blocking IL-4Rα improved lung function by impacting multiple effectors of inflammation and remodeling, except peripheral eosinophil counts, thereby disconnecting blood eosinophils from airway inflammation, remodeling and function. Blocking IL-5 globally reduced eosinophil counts but did not impact inflammatory or functional measures of lung pathology. Whole lung transcriptome analysis revealed that IL-5 or IL-4Rα blockade impacted eosinophil associated genes, whereas IL-4Rα blockade also impacted genes associated with multiple cells, cytokines and chemokines, mucus production, cell:cell adhesion and vascular permeability. CONCLUSIONS: Eosinophils are not the sole contributor to asthma pathophysiology or lung function decline and emphasizes the need to block additional mediators to modify lung inflammation and impact lung function.


Assuntos
Asma , Pneumonia , Animais , Humanos , Camundongos , Asma/metabolismo , Quimiocinas/metabolismo , Citocinas/metabolismo , Células Endoteliais/metabolismo , Inflamação/metabolismo , Interleucina-13/metabolismo , Pulmão/metabolismo , Pneumonia/metabolismo , Interleucina-4/farmacologia
9.
Toxicology ; 484: 153406, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36549504

RESUMO

Environmental pollutants fine particulate matter and di-(2-ethylhexyl) phthalate (DEHP) are believed to be the risk factors for childhood asthma. Allergic asthma is basically an immediate hypersensitivity mediated by IgE, the product of humoral immune response. T follicular helper cells (Tfh) have been newly identified as the crucial T helper cells for supporting B cells to produce immunoglobulins in humoral immunity. Tfh cells are therefore potentially to serve as the diagnostic marker and therapeutic target of immune diseases. In this study, we examined the joint effects of fine particulate matter and DEHP on the initiation and progression of asthma and explored the fundamental role of Tfh cells during the process. Weanling C57BL/6 mice (both sexes) were concurrently exposed to DEHP (intragastric administration at 300 µg/kg) and fine atmospheric particulate matter (mean particle diameter < 4 µm, PM4) (oropharyngeal instillation at 2 mg/kg) once every three days for 30 days (10 times). We found that DEHP displayed adjuvant effects to potentiate PM4 allergen-induced expansion of Tfh and plasma cells, production of serum IgE and IgG1, and occurrence of airway hyper-responsiveness and inflammation. Then PM4 and DEHP co-exposure was performed to Cd4 knock-out mice reconstituted with normal wild-type adoptive Tfh cells or non-Tfh cells. The results of immune adoptive transfusion indicated that the joint immunotoxic effects of PM4 and DEHP were dependent on Tfh cells. We further proved that DEHP could adjuvantly boost PM4-induced expression of BCL-6 and c-MAF and secretion of IL-13 and IL-4 in Tfh cells. In conclusion, these data suggest that DEHP metabolites act in an adjuvant-like manner to aggravate PM4 allergen-induced asthma based on anaphylactic IgE response, resulting from excessive IL-13 and IL-4 synthesized by abnormally differentiated Tfh cells.


Assuntos
Asma , Dietilexilftalato , Masculino , Feminino , Animais , Camundongos , Dietilexilftalato/toxicidade , Células T Auxiliares Foliculares/metabolismo , Interleucina-4/metabolismo , Interleucina-4/farmacologia , Interleucina-13/toxicidade , Interleucina-13/metabolismo , Material Particulado/toxicidade , Camundongos Endogâmicos C57BL , Asma/induzido quimicamente , Asma/metabolismo , Linfócitos T Auxiliares-Indutores , Adjuvantes Imunológicos/farmacologia , Alérgenos/toxicidade , Imunoglobulina E
10.
Food Funct ; 14(1): 413-426, 2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36515134

RESUMO

Salidroside (SAL) is a natural component derived from Rhodiola rosea and is well known for its wide range of biological activities such as its anti-inflammatory and anti-oxidative properties. However, its effects and mechanisms of action related to asthma have not been well explored yet. Recent studies have found that changes in host metabolism are closely related to the progression of asthma. Many natural components can ameliorate asthma by affecting host metabolism. The use of untargeted metabolomics can allow for a better understanding of the metabolic regulatory mechanisms of herbs on asthma. This study aimed to demonstrate the anti-asthmatic effects and metabolic regulatory mechanisms of SAL. In this study, the therapeutic effects of SAL on asthmatic mice were tested at first. Secondly, the effects of SAL on the airway inflammatory reaction, oxidative stress, and airway remodeling were investigated. Finally, untargeted metabolomics analysis was used to explore the influence of SAL on lung metabolites. The results showed that SAL had a significant therapeutic effect on asthmatic model mice. Moreover, SAL treatment lowered interleukin (IL)-4, IL-5, and IL-13 levels but elevated interferon gamma (IFN-γ) and IL-10 levels in bronchoalveolar lavage fluid (BALF). Additionally, it also increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and decreased methane dicarboxylic aldehyde (MDA) levels in the lungs. Besides, SAL-treated mice showed decreased expression of smooth muscle actin (α-SMA), matrix metallopeptidase 2 (MMP2), matrix metallopeptidase 9 (MMP9), and transforming growth factor-beta 1 (TGF-ß1) in the lung. Untargeted metabolomics analysis showed 31 metabolites in the lungs that were influenced by SAL. These metabolites were related to pyrimidine metabolism, steroid hormone biosynthesis, and tricarboxylic acid (TCA) cycle. In conclusion, SAL treatment can reduce the inflammatory response, oxidative stress, and airway remodeling in asthmatic model mice. The mechanism of SAL in the treatment of asthma may be related to the regulation of pyrimidine metabolism, steroid hormone biosynthesis, and the TCA cycle. Further studies can be carried out using targeted metabolomics and in vitro models to deeply elucidate the anti-inflammatory and anti-oxidative mechanisms of SAL on asthma based on regulating metabolism.


Assuntos
Remodelação das Vias Aéreas , Asma , Animais , Camundongos , Asma/induzido quimicamente , Asma/tratamento farmacológico , Asma/metabolismo , Líquido da Lavagem Broncoalveolar , Modelos Animais de Doenças , Hormônios , Pulmão/metabolismo , Metaloproteases/uso terapêutico , Camundongos Endogâmicos BALB C , Ovalbumina , Pirimidinas , Esteroides
11.
Cell Signal ; 102: 110552, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36481410

RESUMO

It is well known that the T Helper (Th)2 bias plays a critical role in allergic asthma. Whereas the Th2 bias is maintained in the local tissues is uncertain. IL-33 is vital for the development of the Th2 polarization. TWIST-1 has an effect on regulating cellular functions. The aberrant activation of RAS sustains certain cellular activities. The aim of this study is to study the role of the interaction between activation of TWIST1 and RAS in inducing and maintaining Th2 polarization in allergic asthma. The epithelial cells of the airways (AEC) were isolated from the broncho-alveolar lavage fluids in patients with asthma. The mediators involved in the over-expression of IL-33 were determined by RNA sequencing. A mouse model was established to test the role of TWIST1 and RAS in developing allergic asthma. We observed a strong expression of TWIST1 in patients with allergic asthma that showed a positive correlation with asthmatic responses. TWIST1 favored the expression of the IL-33 in the AEC. Twist1-deficient AEC-carrying mice did not induce Th2 polarization in the airways. The expression TWIST1 in AECs was positively associated with RAS activation in AECs in patients with allergic asthma. The interaction between RAS and TWIST1 in AECs sustained airway allergic inflammation. Inhibition of TWIST1 or RAS prevented asthma-like inflammation in the mouse airways. In summary, the interaction between TWIST1 and RAS induces and maintains IL-33 expression in AECs to facilitate allergic inflammation in the respiratory tract. Inhibition of TWIST1 or RAS can prevent experimental allergic asthma.


Assuntos
Asma , Interleucina-33 , Animais , Camundongos , Asma/metabolismo , Modelos Animais de Doenças , Células Epiteliais/metabolismo , Inflamação/metabolismo , Interleucina-33/metabolismo , Interleucina-33/farmacologia , Células Th2/metabolismo
12.
Life Sci ; 313: 121289, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36529281

RESUMO

AIMS: Augmented smooth muscle contractility of the airways associated with an increased expression of RhoA, a monomeric GTPase responsible for Ca2+ sensitization of contraction, is one of the causes of airway hyperresponsiveness. However, the mechanism of the altered properties of airway smooth muscle cells, including the RhoA upregulation, is not fully understood. This study aims to define functional role of a long non-coding RNA MALAT1 in the RhoA expression and development of bronchial smooth muscle (BSM) hyper-contractility. MAIN METHODS: Cultured human BSM cells were transfected with MALAT1 antisense oligonucleotide (AS), miR-133a-3p mimic, and/or inhibitor, and then stimulated with interleukin-13 (IL-13). In animal experiments, the ovalbumin (OA)-sensitized mice were repeatedly challenged with aerosolized OA to induce asthmatic reaction. KEY FINDINGS: Treatment of the cells with IL-13 induced an increase in RhoA protein. Either MALAT1 AS or miR-133a-3p mimic transfection inhibited the IL-13-induced upregulation of RhoA. The inhibitory effect of MALAT1 AS was abolished by co-transfection with miR-133a-3p inhibitor. In BSMs of the murine asthma model, upregulations of Malat1 and RhoA protein were observed concomitantly with downregulation of miR-133a-3p. SIGNIFICANCE: These findings suggest that MALAT1 positively regulates RhoA protein expression by inhibiting miR-133a-3p in BSM cells, and that its upregulation causes the RhoA upregulation, resulting in an augmented BSM contractility.


Assuntos
Asma , Hiper-Reatividade Brônquica , MicroRNAs , RNA Longo não Codificante , Humanos , Animais , Camundongos , RNA Longo não Codificante/metabolismo , Proteína rhoA de Ligação ao GTP/genética , Proteína rhoA de Ligação ao GTP/metabolismo , Interleucina-13/metabolismo , Hiper-Reatividade Brônquica/metabolismo , Brônquios/metabolismo , Asma/metabolismo , Miócitos de Músculo Liso/metabolismo , MicroRNAs/metabolismo
13.
Life Sci ; 313: 121304, 2023 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-36535402

RESUMO

AIMS: Adhesion molecules play vital roles in the induction of airway hyperresponsiveness (AHR) or airway inflammation. The down-regulation of catenin alpha-like 1 (CTNNAL1) in the bronchial epithelial cells of asthma patients and mice models has been noted in our previous study. In this work, we further explore the underlying mechanism of CTNNAL1 in asthma. MAIN METHODS: We constructed a house dust mite (HDM)-induced asthma animal model on control mice and applied CTNNAL1-siRNA transfection to create CTNNAL1-deficient mice. KEY FINDINGS: We documented much more severe airway inflammation and increased leukocyte infiltration in the lungs of the CTNNAL1-deficient mice comparing to control mice, along with elevated expression of inflammatory cytokines. Dexamethasone (DEX) treatment led to less reduced inflammation in CTNNAL1-deficient mice compared with control mice. Immunoprecipitation confirmed the interaction between heat shock protein90 (hsp90) and CTNNAL1. The expression of hsp90 was upregulated after CTNNAL1 silencing. Meanwhile, the use of hsp90 inhibitor geldanamycin significantly decreased the expression of NR3C1, ICAM-1 and the ratio of p-p65/p65 in CTNNAL1-silenced 16HBE14o- cells. Both geldanamycin and DEX could function to suppress the expression of ICAM-1 and the phosphorylation level of p65. Nevertheless, the anti-inflammatory effect of DEX proved less potent than geldanamycin in the CTNNAL1-silenced group. The combined therapy of geldanamycin and DEX significantly decreased the inflammatory responses in CTNNAL1-deficient HBE cells than DEX monotherapy. SIGNIFICANCE: Our study corroborates that CTNNAL1 deficiency induced aggravated airway inflammation and rendered insensitivity to glucocorticoids via triggering hsp90 signaling pathway.


Assuntos
Asma , Glucocorticoides , Camundongos , Animais , Glucocorticoides/farmacologia , Glucocorticoides/metabolismo , Molécula 1 de Adesão Intercelular/genética , Molécula 1 de Adesão Intercelular/metabolismo , Asma/metabolismo , Pulmão/metabolismo , Transdução de Sinais , Resposta ao Choque Térmico , Inflamação/metabolismo , Pyroglyphidae , Modelos Animais de Doenças , alfa Catenina/genética , alfa Catenina/metabolismo
14.
Sci Rep ; 12(1): 22476, 2022 12 28.
Artigo em Inglês | MEDLINE | ID: mdl-36577785

RESUMO

Eosinophils are granulocytes that play a significant role in the pathogenesis of asthma and other airway diseases. Directing patient treatment based on the level of eosinophilia has been shown to be extremely effective in reducing exacerbations and therefore has tremendous potential as a routine clinical test. Herein, we describe the in vitro selection and optimization of DNA aptamers that bind to eosinophil peroxidase (EPX), a protein biomarker unique to eosinophils. Fifteen rounds of magnetic bead aptamer selection were performed prior to high throughput DNA sequencing. The top 10 aptamer candidates were assessed for EPX binding using a mobility shift assay. This process identified a lead aptamer candidate termed EAP1-05 with low nanomolar affinity and high specificity for EPX over other common sputum proteins. This aptamer sequence was further optimized through truncation and used to develop an easy-to-use colourimetric pull-down assay that can detect EPX over a concentration range from 1 - 100 nM in processed sputum. Forty-six clinical samples were processed using a new sputum dispersal method, appropriate for a rapid assessment assay, that avoids centrifugation and lengthy processing times. The assay showed 89% sensitivity and 96% specificity to detect eosinophilia (compared to gold standard sputum cytometry), with results being produced in under an hour. This assay could allow for an easy assessment of eosinophil activity in the airway to guide anti-inflammatory therapy for several airway diseases.


Assuntos
Asma , Eosinofilia , Humanos , Peroxidase de Eosinófilo/metabolismo , Escarro/metabolismo , Eosinofilia/patologia , Eosinófilos/metabolismo , Asma/metabolismo
15.
Int J Mol Sci ; 23(23)2022 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-36499612

RESUMO

Classically, the effects elicited by corticosteroids (CS) are mediated by the binding and activation of cytosolic glucocorticoid receptors (GR). However, several of the non-genomic effects of CS seem to be mediated by putative non-classic membrane receptors characterized by pharmacological properties that are different from those of classic cytosolic GR. Since pre-clinical findings suggest that inhaled CS (ICS) may also regulate the bronchial contractile tone via putative CS membrane-associate receptors, the aim of this review was to systematically report and discuss the impact of CS on human airway smooth muscle (ASM) contractility and airway hyperresponsiveness (AHR). Current evidence indicates that CS have significant genomic/non-genomic beneficial effects on human ASM contractility and AHR, regardless of their anti-inflammatory effects. CS are effective in reducing either the expression, synthesis or activity of α-actin, CD38, inositol phosphate, myosin light chain kinase, and ras homolog family member A in response to several pro-contractile stimuli; overall these effects are mediated by the genomic action of CS. Moreover, CS elicited a strong bronchorelaxant effect via the rapid activation of the Gsα-cyclic-adenosine-monophosphate-protein-kinase-A pathway in hyperresponsive airways. The possibility of modulating the dose of the ICS in a triple ICS/long-acting ß2-adrenoceptor agonist/long-acting muscarinic antagonist fixed-dose combination supports the use of a Triple MAintenance and Reliever Therapy (TriMART) in those asthmatic patients at Step 3-5 who may benefit from a sustained bronchodilation and have been suffering from an increased parasympathetic tone.


Assuntos
Asma , Músculo Liso , Humanos , Músculo Liso/metabolismo , Asma/metabolismo , Brônquios/metabolismo , Contração Muscular/fisiologia , Corticosteroides/uso terapêutico
16.
Respir Res ; 23(1): 381, 2022 Dec 28.
Artigo em Inglês | MEDLINE | ID: mdl-36578010

RESUMO

BACKGROUND: Airway fibrosis is one of the pathological characteristics of severe asthma. Transforming growth factor (TGF)-ß has been known to promote epithelial-mesenchymal transition formation and to play a role in the progression of tissue fibrosis. Cellular communication network factor 2 (CCN2) and fibronectin (FN) are well-known markers of EMT and fibrosis. However, whether AREG is involved in TGF-ß-induced CCN2 and FN expression in human lung epithelial cells is unknown. METHODS: AREG and FN were analyzed by immunofluorescence staining on ovalbumin-challenged mice. CCN2 and FN expression were evaluated in human lung epithelial (A459) cells following TGF or AREG treatment for the indicated times. Secreted AREG from A549 cells was detected by ELISA. Cell migration was observed by a wound healing assay. Chromatin immunoprecipitation was used to detect the c-Jun binding to the CCN2 promoter. RESULTS: AREG and FN expression colocalized in lung tissues from mice with ovalbumin-induced asthma by immunofluorescence staining. Moreover, TGF-ß caused the release of AREG from A549 cells into the medium. Smad3 siRNA down-regulated AREG expression. AREG also stimulated CCN2 and FN expression, JNK and c-Jun phosphorylation, and cell migration in A549 cells. AREG small interfering (si) RNA inhibited TGF-ß-induced expression of CCN2, FN, and cell migration. Furthermore, AREG-induced CCN2 and FN expression were inhibited by EGFR siRNA, a JNK inhibitor (SP600125), and an activator protein-1 (AP-1) inhibitor (curcumin). EGFR siRNA attenuated AREG-induced JNK and c-Jun phosphorylation. Moreover, SP600125 downregulated AREG-induced c-Jun phosphorylation. CONCLUSION: These results suggested that AREG mediates the TGF-ß-induced EMT in human lung epithelial cells through EGFR/JNK/AP-1 activation. Understanding the role of AREG in the EMT could foster the development of therapeutic strategies for airway remodeling in severe asthma.


Assuntos
Asma , Fator de Crescimento Transformador beta , Humanos , Camundongos , Animais , Fator de Crescimento Transformador beta/metabolismo , Anfirregulina/genética , Anfirregulina/metabolismo , Fibronectinas/metabolismo , Ovalbumina/toxicidade , Fator de Transcrição AP-1/metabolismo , Pulmão/metabolismo , Células Epiteliais/metabolismo , Transição Epitelial-Mesenquimal , Asma/metabolismo , Receptores ErbB/metabolismo , RNA Interferente Pequeno/metabolismo , Fibrose , Fator de Crescimento Transformador beta1/farmacologia
17.
Front Immunol ; 13: 1015529, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36518751

RESUMO

Background: Neutrophils are involved in the pathophysiology of allergic asthma, where the Eosinophil Cationic Protein (ECP) is a critical inflammatory mediator. Although ECP production is attributed to eosinophils, we reported that ECP is also present in neutrophils from allergic patients where, in contrast to eosinophils, it is produced in an IgE-dependent manner. Given the key role of ECP in asthma, we investigated the molecular mechanisms involved in ECP production as well as the effects induced by agonists and widely used clinical approaches. We also analyzed the correlation between ECP production and lung function. Methods: Neutrophils from allergic asthmatic patients were challenged with allergens, alone or in combination with cytokines, in the presence of cell-signaling inhibitors and clinical drugs. We analyzed ECP levels by ELISA and confocal microscopy. Lung function was assessed by spirometry. Results: IgE-mediated ECP release is dependent on phosphoinositide 3-kinase, the extracellular signal-regulated kinase (ERK1/2) and the production of reactive oxygen species by NADPH-oxidase. Calcineurin phosphatase and the transcription factor NFAT are also involved. ECP release is enhanced by the cytokines interleukin (IL)-5 and granulocyte macrophage-colony stimulating factor, and inhibited by interferon-γ, IL-10, clinical drugs (formoterol, tiotropium and budesonide) and allergen-specific IT. We also found an inverse correlation between asthma severity and ECP levels. Conclusions: Our results suggest the molecular pathways involved in ECP production and potential therapeutic targets. We also provide a new method to evaluate disease severity in asthmatic patients based on the quantification of in vitro ECP production by peripheral neutrophils.


Assuntos
Asma , Hipersensibilidade , Humanos , Proteína Catiônica de Eosinófilo/metabolismo , Neutrófilos/metabolismo , Fosfatidilinositol 3-Quinases , Alérgenos , Asma/tratamento farmacológico , Asma/metabolismo , Citocinas/metabolismo , Imunoglobulina E
18.
Front Immunol ; 13: 973673, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36479132

RESUMO

Asthmatics are more susceptible to viral infections than healthy individuals and are known to have impaired innate anti-viral defences. Influenza A virus causes significant morbidity and mortality in this population. Immuno-modulatory regulators (IMRs) such as PD-1 are activated on T cells following viral infection as part of normal T cell activation responses, and then subside, but remain elevated in cases of chronic exposure to virus, indicative of T cell exhaustion rather than activation. There is evidence that checkpoint inhibition can enhance anti-viral responses during acute exposure to virus through enhancement of CD8+T cell function. Although elevated PD-1 expression has been described in pulmonary tissues in other chronic lung diseases, the role of IMRs in asthma has been relatively unexplored as the basis for immune dysfunction. We first assessed IMR expression in the peripheral circulation and then quantified changes in IMR expression in lung tissue in response to ex-vivo influenza infection. We found that the PD-1 family members are not significantly altered in the peripheral circulation in individuals with severe asthma but are elevated in pulmonary tissues following ex-vivo influenza infection. We then applied PD-1 Mab inhibitor treatment to bronchial biopsy tissues infected with influenza virus and found that PD-1 inhibition was ineffective in asthmatics, but actually increased infection rates in healthy controls. This study, therefore, suggests that PD-1 therapy would not produce harmful side-effects when applied in people with severe asthma, but could have important, as yet undescribed, negative effects on anti-viral responses in healthy individuals that warrant further investigation.


Assuntos
Asma , Influenza Humana , Receptor de Morte Celular Programada 1 , Humanos , Influenza Humana/complicações , Receptor de Morte Celular Programada 1/antagonistas & inibidores , Receptor de Morte Celular Programada 1/metabolismo , Asma/metabolismo , Asma/virologia , Progressão da Doença , Linfócitos T CD8-Positivos
19.
Can Respir J ; 2022: 1485719, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36582191

RESUMO

Asthma is a chronic inflammatory disturbance of the airways in which many cells and cellular elements are involved. Wheezing, breathlessness, chest tightness, and coughing, especially at night or in the early morning, are typical symptoms of asthma. At present, inhaled corticosteroid (ICS) and long-acting ß-agonists (LABAs) are standard treatments for regular management. Oral corticosteroids (OCSs) were recommended for controlling asthma exacerbation but only for a short-term treatment because of the side effects on organs. Biologic therapies have achieved exciting and notable effects in clinical treatment but are not applicable for all phenotypes of asthma. At present, some new approaches are under exploration to lessen side effects and improve curative effects. Studies have revealed that bone marrow mesenchymal stem cells (BMMSCs) hold various curative effects in asthma and may benefit in the long term with high safety. Extracellular vesicles (EVs) enriched in body fluid were characterized as subcomponents of extracellular vesicles and delivered carriers combined with genetic messages in vivo. The therapeutic potential of exosomes has become a research hotspot in many diseases. BMMSC-derived exosomes were considered as the dominant part of BMMSCs in cell-to-cell communications and playing curative effects. Points also hold that BMMSC-Exo could interfere with airway inflammation and airway remolding in asthma via modulating the immune response, regulating gene expression, adjusting the phenotype of macrophage, etc. However, BMMSC-Exo still lacked more clinical trials for evaluating the effects on asthma, and the technology of extraction and purification still needs to be improved for wide use. This review aims to draw the relationship among asthma, BMMSC, and exosome, which may provide innovate ideas for treatment of asthma, and arouse attention about the curative potential of BMMSC-Exo.


Assuntos
Asma , Exossomos , Células-Tronco Mesenquimais , Humanos , Asma/tratamento farmacológico , Asma/metabolismo , Corticosteroides/uso terapêutico , Quimioterapia Combinada , Células-Tronco Mesenquimais/metabolismo
20.
Respir Res ; 23(1): 378, 2022 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-36572876

RESUMO

BACKGROUND: To date, reliable biomarkers for asthma have not been identified. MicroRNAs (miRNAs) are small, non-coding RNAs that negatively regulate post-transcriptional gene expression, and they are involved in various diseases, including asthma. MiRNAs may serve as ideal biomarkers due to their ability to regulate multiple pathways. This study aims to identify miRNA biomarker signatures for asthma. METHODS: We used the house dust mite (HDM) mouse model of allergic inflammation. Mice were phenotyped by assessing lung function, allergic response, airway inflammation, and remodeling. The miRNA signature profiles in serum and lung tissue were determined by small RNA sequencing, and data were analyzed using Qiagen CLC Genomics Workbench. To identify relevant gene targets, we performed mRNA sequencing, followed by miRNA-targets analysis. These miRNAs and targets were subject to subsequent pathway and functional analyses. RESULTS: Mice exposed to HDM developed phenotypic features of allergic asthma. miRNA sequencing analysis showed that 213 miRNAs were substantially dysregulated (FDR p-value < 0.05 and fold change expression > + 1.5 and < - 1.5) in the lung of HDM mice relative to the control mice. In contrast, only one miRNA (miR-146b-5p) was significantly increased in serum. Target analysis of lung dysregulated miRNAs revealed a total of 131 miRNAs targeting 211 mRNAs. Pathway analysis showed T helper 2/1 (Th2/Th1) as the top significantly activated signaling pathway associated with the dysregulated miRNAs. The top enriched diseases were inflammatory response and disease, which included asthma. Asthma network analysis indicated that 113 of 131 miRNAs were directly associated with asthma pathogenesis. CONCLUSIONS: These findings suggest that most dysregulated miRNAs in the HDM model were associated with asthma pathogenesis via Th2 signaling. We identified a panel of 30 miRNAs as potential biomarker candidates for asthma.


Assuntos
Asma , MicroRNAs , Animais , Camundongos , MicroRNAs/genética , Redes Reguladoras de Genes , Asma/diagnóstico , Asma/genética , Asma/metabolismo , Biomarcadores , Pyroglyphidae , Inflamação
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