RÉSUMÉ
BACKGROUND: The retinal vasculature, a crucial component of the human body, mirrors various illnesses such as cardiovascular disease, glaucoma, and retinopathy. Accurate segmentation of retinal vessels in funduscopic images is essential for diagnosing and understanding these conditions. However, existing segmentation models often struggle with images from different sources, making accurate segmentation in crossing-source fundus images challenging. METHODS: To address the crossing-source segmentation issues, this paper proposes a novel Multi-level Adversarial Learning and Pseudo-label Denoising-based Self-training Framework (MLAL&PDSF). Expanding on our previously proposed Multiscale Context Gating with Breakpoint and Spatial Dual Attention Network (MCG&BSA-Net), MLAL&PDSF introduces a multi-level adversarial network that operates at both the feature and image layers to align distributions between the target and source domains. Additionally, it employs a distance comparison technique to refine pseudo-labels generated during the self-training process. By comparing the distance between the pseudo-labels and the network predictions, the framework identifies and corrects inaccuracies, thus enhancing the accuracy of the fine vessel segmentation. RESULTS: We have conducted extensive validation and comparative experiments on the CHASEDB1, STARE, and HRF datasets to evaluate the efficacy of the MLAL&PDSF. The evaluation metrics included the area under the operating characteristic curve (AUC), sensitivity (SE), specificity (SP), accuracy (ACC), and balanced F-score (F1). The performance results from unsupervised domain adaptive segmentation are remarkable: for DRIVE to CHASEDB1, results are AUC: 0.9806, SE: 0.7400, SP: 0.9737, ACC: 0.9874, and F1: 0.8851; for DRIVE to STARE, results are AUC: 0.9827, SE: 0.7944, SP: 0.9651, ACC: 0.9826, and F1: 0.8326. CONCLUSION: These results demonstrate the effectiveness and robustness of MLAL&PDSF in achieving accurate segmentation results from crossing-domain retinal vessel datasets. The framework lays a solid foundation for further advancements in cross-domain segmentation and enhances the diagnosis and understanding of related diseases.
Sujet(s)
Vaisseaux rétiniens , Humains , Vaisseaux rétiniens/imagerie diagnostique , Traitement d'image par ordinateur/méthodes , AlgorithmesRÉSUMÉ
Objective: Asthma, a chronic inflammatory disease in which type 2 T helper cells (Th2) play a causative role in the development of T2 asthma. N6-methyladenosine (m6A) modification, an mRNA modification, and methyltransferase-like 3 (METTL3) is involved in the development of T2 asthma by inhibiting Th2 cell differentiation. Sex determining region Y-box protein 5 (SOX5) is involved in regulating T cell differentiation, but its role in T2 asthma was unclear. The objective of this study was to explore the role of METTL3 and SOX5 in T2 asthma and whether there is an interaction between the two. Materials and methods: Adults diagnosed with T2 asthma (n = 14) underwent clinical information collection and pulmonary function tests. In vivo and in vitro T2 asthma models were established using female C57BL/6 mice and human bronchial epithelial cells (HBE). The expressions of METTL3 and SOX5 were detected by Western blot and qRT-PCR and Western blot. Th2 cell differentiation was determined by flow cytometry and IL-4 level was detected by ELISA. m6A methylation level was determined by m6A quantitative assay. The relationship between METTL3 expression and clinical parameters was determined by Spearman rank correlation analysis. The function of METTL3 and SOX5 genes in asthma was investigated in vitro and in vivo. The RNA immunoprecipitation assay detected the specific interaction between METTL3 and SOX5. Results: Patients with T2 asthma displayed lower METTL3 levels compared to healthy controls. Within this group, a negative correlation was observed between METTL3 and Th2 cells, while a positive correlation was noted between METTL3 and clinical parameters as well as Th1 cells. In both in vitro and in vivo models representing T2 asthma, METTL3 levels decreased significantly, while SOX5 levels showed the opposite trend. Overexpression of METTL3 gene in HBE cells significantly inhibited Th2 cell differentiation and increased m6A methylation activity. From a mechanism perspective, low METTL3 negatively regulates SOX5 expression through m6A modification dependence, while high SOX5 expression is positively associated with T2 asthma severity. Cell transfection experiments confirmed that METTL3 regulates Th2 cell differentiation and IL-4 release through SOX5. Conclusions: Overall, our results indicate that METTL3 alleviates Th2 cell differentiation in T2 asthma by modulating the m6A methylation activity of SOX5 in bronchial epithelial cells. This mechanism could potentially serve as a target for the prevention and management of T2 asthma.
RÉSUMÉ
BACKGROUND: Allergic asthma is a heterogeneous disease and new strategies are needed to prevent or treat this disease. Studies have shown that probiotic interventions are effective in preventing asthma. Here, we investigated the impact of Saccharomyces boulardii (S. boulardii) on ovalbumin (OVA)-induced allergic asthma in mice, as well as the underlying mechanisms. METHODS: First, we constructed a mouse asthma model using OVA and given S. boulardii intervention. Next, we measured N6-methyladenosine (m6A) levels in lung injury tissues. 16 s rRNA was employed to identify different gut microbiota in fecal samples. The analysis of differential metabolites in feces was performed by non-targeted metabolomics. Pearson correlation coefficient was utilized to analyze correlation between gut microbiota, metabolites and methyltransferase-like 3 (METTL3). Finally, we collected mouse feces treated by OVA and S. boulardii intervention for fecal microbiota transplantation (FMT) and interfered with METTL3. RESULTS: S. boulardii improved inflammation and oxidative stress and alleviated lung damage in asthmatic mice. In addition, S. boulardii regulated m6A modification levels in asthmatic mice. 16 s rRNA sequencing showed that S. boulardii remodeled gut microbiota homeostasis in asthmatic mice. Non-targeted metabolomics analysis showed S. boulardii restored metabolic homeostasis in asthmatic mice. There was a correlation between gut microbiota, differential metabolites, and METTL3 analyzed by Pearson correlation. Additionally, through FMT and interference of METTL3, we found that gut microbiota mediated the up-regulation of METTL3 by S. boulardii improved inflammation and oxidative stress in asthmatic mice, and alleviated lung injury. CONCLUSIONS: S. boulardii alleviated allergic asthma by restoring gut microbiota and metabolic homeostasis via up-regulation of METTL3 in an m6A-dependent manner.
Sujet(s)
Adénosine , Asthme , Modèles animaux de maladie humaine , Microbiome gastro-intestinal , Homéostasie , Methyltransferases , Probiotiques , Saccharomyces boulardii , Régulation positive , Animaux , Asthme/thérapie , Asthme/métabolisme , Asthme/immunologie , Asthme/étiologie , Asthme/microbiologie , Methyltransferases/métabolisme , Methyltransferases/génétique , Microbiome gastro-intestinal/immunologie , Souris , Adénosine/métabolisme , Adénosine/analogues et dérivés , Probiotiques/administration et posologie , Probiotiques/usage thérapeutique , Femelle , Transplantation de microbiote fécal , Ovalbumine/immunologie , Souris de lignée BALB CRÉSUMÉ
Cardiovascular diseases are the leading cause of mortality, and accurate segmentation of ventricular regions incardiac magnetic resonance images (MRIs) is crucial for diagnosing and treating these diseases. However, fully automated and accurate right ventricle (RV) segmentation remains challenging due to the irregular cavities with ambiguous boundaries and mutably crescentic structures with relatively small targets of the RV regions in MRIs. In this article, a triple-path segmentation model, called FMMsWC, is proposed by introducing two novel image feature encoding modules, i.e., the feature multiplexing (FM) and multiscale weighted convolution (MsWC) modules, for the RV segmentation in MRIs. Considerable validation and comparative experiments were conducted on two benchmark datasets, i.e., the MICCAI2017 Automated Cardiac Diagnosis Challenge (ACDC), and the Multi-Centre, Multi-Vendor & Multi-Disease Cardiac Image Segmentation Challenge (M&MS) datasets. The FMMsWC outperforms state-of-the-art approaches, and its performance can approach that of the manual segmentation results by clinical experts, facilitating accurate cardiac index measurement for the rapid assessment of cardiac function and aiding diagnosis and treatment of cardiovascular diseases, which has great potential for clinical applications.
Sujet(s)
Maladies cardiovasculaires , Ventricules cardiaques , Humains , Ventricules cardiaques/imagerie diagnostique , IRM dynamique/méthodes , Imagerie par résonance magnétique/méthodes , Coeur , Traitement d'image par ordinateur/méthodesRÉSUMÉ
OBJECTIVES: Obstructive sleep apnea hypopnea syndrome (OSAHS) is a common disease that seriously affects health. Continuous positive airway pressure (CPAP) therapy is the preferred treatment for moderate-to-severe OSAHS patients. However, poor adherence to CPAP is a major obstacle in the treatment of OSAHS. Information-motivation-behavioral (IMB) skills, as a kind of mature technology to change the behavior, has been used in various health areas to improve treatment adherence. This study aims to explore the effects of the IMB skills intervention on CPAP adherence in OSAHS patients. METHODS: Patients who were primary diagnosed with moderate-to-severe OSAHS were randomly divided into the IMB group (n=62) and the control group (n=58). The patients in the IMB group received CPAP therapy and the IMB skills intervention for 4 weeks. The patients in the control group received CPAP therapy and a usual health care provided by a registered nurse. We collected the baseline data of the general information, including age, sex, body mass index (BMI), the Epworth Sleepiness Scale (ESS) score, the Hospital Anxiety and Depression Scale (HADS) score, and indicators about disease severity [apnea-hypopnea index (AHI), percentage of time with arterial oxygen saturation SaO2<90% (T90), average SaO2, lowest SaO2, arousal index]. After CPAP titration, we collected CPAP therapy-relevant parameters (optimal pressure, maximum leakage, average leakage, 95% leakage, and residual AHI), score of satisfaction and acceptance of CPAP therapy, and score of willingness to continue CPAP therapy. After 4 weeks treatment, we collected the ESS score, HADS score, CPAP therapy-relevant parameters, effective CPAP therapy time per night, CPAP therapy days within 4 weeks, CPAP adherence rate, score of satisfaction and acceptance of CPAP therapy, and score of willingness to continue CPAP therapy. Visual analog scale (VAS) of 0-5 was used to evaluate the satisfaction and acceptance of IMB intervention measures in the IMB group. RESULTS: There were no significant differences in the baseline level of demographic parameters, ESS score, HADS score, disease severity, and CPAP therapy related parameters between the IMB group and the control group (all P>0.05). There were no significant differences in score of willingness to continue CPAP therapy, as well as score of satisfaction and acceptance of CPAP therapy after CPAP titration between the IMB group and the control group (both P>0.05). After 4 weeks treatment, the ESS score, HADS score, maximum leakage, average leakage, and 95% leakage of the IMB group were significantly decreased, while the score of satisfaction and acceptance of CPAP therapy and willingness to continue CPAP therapy of the IMB group were significantly increased (all P<0.05); while the above indexes in the control group were not different before and after 4 weeks treatment (all P<0.05). Compared with the control group, the ESS score, HADS score, maximum leakage, average leakage, and 95% leakage of the IMB group after 4 weeks treatment were significantly lower (all P<0.05); the effective CPAP therapy time, CPAP therapy days within 4 weeks, score of satisfaction and acceptance of CPAP therapy, score of willingness to continue CPAP therapy of the IMB group were significantly higher (all P<0.05). The rate of CPAP therapy adherence in 4 weeks of the IMB group was significantly higher than that of the control group (90.3% vs 62.1%, P<0.05). The VAS of overall satisfaction with IMB skills intervention measures was 4.46±0.35. CONCLUSIONS: IMB skills intervention measures can effectively improve the adherence of CPAP therapy in OSAHS patients, and is suitable for clinical promotion.
Sujet(s)
Ventilation en pression positive continue , Syndrome d'apnées obstructives du sommeil , Humains , Motivation , Oxymétrie , Observance par le patient , Syndrome d'apnées obstructives du sommeil/diagnostic , SyndromeRÉSUMÉ
Accurate cardiac segmentation of multimodal images, e.g., magnetic resonance (MR), computed tomography (CT) images, plays a pivot role in auxiliary diagnoses, treatments and postoperative assessments of cardiovascular diseases. However, training a well-behaved segmentation model for the cross-modal cardiac image analysis is challenging, due to their diverse appearances/distributions from different devices and acquisition conditions. For instance, a well-trained segmentation model based on the source domain of MR images is often failed in the segmentation of CT images. In this work, a cross-modal images-oriented cardiac segmentation scheme is proposed using a symmetric full convolutional neural network (SFCNN) with the unsupervised multi-domain adaptation (UMDA) and a spatial neural attention (SNA) structure, termed UMDA-SNA-SFCNN, having the merits of without the requirement of any annotation on the test domain. Specifically, UMDA-SNA-SFCNN incorporates SNA to the classic adversarial domain adaptation network to highlight the relevant regions, while restraining the irrelevant areas in the cross-modal images, so as to suppress the negative transfer in the process of unsupervised domain adaptation. In addition, the multi-layer feature discriminators and a predictive segmentation-mask discriminator are established to connect the multi-layer features and segmentation mask of the backbone network, SFCNN, to realize the fine-grained alignment of unsupervised cross-modal feature domains. Extensive confirmative and comparative experiments on the benchmark Multi-Modality Whole Heart Challenge dataset show that the proposed model is superior to the state-of-the-art cross-modal segmentation methods.
Sujet(s)
Traitement d'image par ordinateur , , Attention , Coeur/imagerie diagnostique , Humains , Imagerie par résonance magnétiqueRÉSUMÉ
METHODS: Ovalbumin was used to induce allergic asthma following administration of YFP for one week in mice, to collect the lung tissues, bronchoalveolar lavage fluid (BLFA), and feces. The pathological state, tight-junction proteins, inflammatory and oxidative stress-associated biomarkers, and TLRs/NF-κB signaling pathway of the lung tissues were evaluated by HE staining, immunofluorescence, ELISA, and WB, separately. RT-PCR was used to test oxidative stress-associated genes. Leukocyte counts of BLFA and intestinal microbiota were also analyzed using a hemocytometer and 16S rDNA-sequencing, separately. RESULT: YFP ameliorated the lung injury of the mouse asthma model by inhibiting peribronchial and perivascular infiltrations of eosinophils and increasing tight-junction protein expression. YFP inhibited the decrease in the number of BALF leukocytes and expression of inflammatory-related genes and reversed OVA-induced TLRs/NF-κB signaling pathway activation. YFP ameliorated the level of oxidative stress in the lung of the mouse asthma model by inhibiting MDA and promoting the protein level of GSH-PX, SOD, CAT, and oxidative-related genes. ATG5, Beclin1, and LC3BII/I were significantly upregulated in asthma mice, which were greatly suppressed by the introduction of YFP, indicating that YFP ameliorated the autophagy in the lung of the mouse asthma model. Lastly, the distribution of bacterial species was slightly changed by YFP in asthma mice, with a significant difference in the relative abundance of 6 major bacterial species between the asthma and YFP groups. CONCLUSION: Our research showed that YFP might exert antiasthmatic effects by inhibiting airway allergic inflammation and oxidative stress level through suppressing autophagy.
Sujet(s)
Autophagie , Liquide de lavage bronchoalvéolaire/microbiologie , Fermentation , Stress oxydatif , Prébiotiques , Levures/métabolisme , Animaux , Antiasthmatiques/pharmacologie , Asthme/métabolisme , Marqueurs biologiques/métabolisme , ADN ribosomique/métabolisme , Inflammation , Poumon/métabolisme , Mâle , Souris , Souris de lignée BALB C , Analyse de séquence d'ADN , Transduction du signal/effets des médicaments et des substances chimiquesRÉSUMÉ
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the cause of the outbreak of coronavirus disease 2019 in Wuhan City, China. The SARS-CoV-2 is genetically similar to the coronavirus derived from bat. The SARS-CoV-2, the SARS-CoV and the Middle East respiratory syndrome coronavirus (MERS-CoV) all belong to beta coronavirus. Since the outbreak of the coronavirus disease 2019, effective antiviral drugs have become a hot issue in the world. Very little about SARS-CoV-2 is known and there is no precedent for treatment. The National Health Commission has repeatedly revised the diagnosis and treatment guide for the coronavirus disease 2019. The latest guide is "New Coronary Virus-Infected Pneumonia Diagnosis and Treatment Plan (Seventh Trial Version)"(short for Seventh Version of Diagnosis and Treatment Plan). But the use of antiviral drugs is still on trial and no rigorous clinical trials data is available. Hot anti-SARS-CoV-2 drugs include interferon α, ribavirin, lopinavir/ritonavir, chloroquine phosphate, abidol, as well as hydroxychloroquine sulfate and remdesivir. But the later 2 drugs aren't mentioned in the Seventh Version of Diagnosis and Treatment Plan.
Sujet(s)
Antiviraux/usage thérapeutique , Infections à coronavirus/traitement médicamenteux , Pneumopathie virale/traitement médicamenteux , Betacoronavirus , COVID-19 , Chine , Humains , Pandémies , Guides de bonnes pratiques cliniques comme sujet , SARS-CoV-2 , Traitements médicamenteux de la COVID-19RÉSUMÉ
Anaplastic lymphoma kinase (ALK) fusion events account for ~3-7% genetic alterations in patients with non-small cell lung cancer (NSCLC). In this study, we identified the ALK fusion patterns and a novel ALK fusion partner in 44 ALK positive NSCLC patients using a customized HapOncoCDx panel, and identified ALK fusion partners. The most common partner is EML4, forming the variant 1 (v1, E13:A20, 18/44), variant 2 (v2, E20:A20, 5/44), and variant 3 (v3, E6:A20, 13/44). Moreover, we detected a new ALK fusion partner HMBOX1. At the mutation level, TP53 is the most frequently mutated gene (24%), followed by ALK (12%) and STED2 (12%). The median tumor mutation burden (TMB) of these samples is 2.29 mutations/Mb, ranging from 0.76 mut/Mb to 16.79 muts/Mb. We further elaborately portrayed the TP53 mutation sites on the peptide sequence of the encoded protein by lollipop. The mutational signature and copy number alterations (CNAs) of the samples were also analyzed. The CNA events were found in 13 (13/44) patients, and the most commonly amplified genes were MDM2 (n = 4/13) and TERT (n = 4/13). Together, these results may guide personalized clinical management of patients with ALK fusion in the era of precision medicine.
RÉSUMÉ
Accurate segmentation of brain tumors from magnetic resonance (MR) images play a pivot role in assisting diagnoses, treatments and postoperative evaluations. However, due to its structural complexities, e.g., fuzzy tumor boundaries with irregular shapes, accurate 3D brain tumor delineation is challenging. In this paper, an intersection over union (IOU) constraint 3D symmetric full convolutional neural network (IOUC-3DSFCNN) model fused with multimodal auto-context is proposed for the 3D brain tumor segmentation. IOUC-3DSFCNN incorporates 3D residual groups into the classic 3DU-Net to further deepen the network structure to obtain more abstract voxel features under a five-layer cohesion architecture to ensure the model stability. The IOU constraint is used to address the issue of extremely unbalanced tumor foreground and background regions in MR images. In addition, to obtain more comprehensive and stable 3D brain tumor profiles, the multimodal auto-context information is fused into the IOUC-3DSFCNN model to achieve end-to-end 3D brain tumor profiles. Extensive confirmatory and comparative experiments conducted on the benchmark BRATS 2017 dataset demonstrate that the proposed segmentation model is superior to classic 3DU-Net-relevant and other state-of-the-art segmentation models, which can achieve accurate 3D tumor profiles on multimodal MRI volumes even with blurred tumor boundaries and big noise.
Sujet(s)
Tumeurs du cerveau/imagerie diagnostique , Apprentissage profond , Imagerie tridimensionnelle/méthodes , Imagerie par résonance magnétique/méthodes , Jeux de données comme sujet , Humains , Modèles statistiques , Reproductibilité des résultatsRÉSUMÉ
We report a case of H7N9 avian influenza pneumonia in a pregnant woman who developed acute respiratory distress syndrome (ARDS) managed with extracorporeal membrane oxygenation (ECMO). A 29-year-old, 27 week pregnant woman developed rapidly progressive pneumonia with bilateral infiltrates on chest x-ray and was confirmed to have influenza A (H7N9) infection. Her condition deteriorated and she developed ARDS which was managed with veno-venous extracorporeal membrane oxygenation (V-V ECMO) and treated with antimicrobials. Her clinical symptoms and oxygenation gradually improved and the ECMO was discontinued on the 19t day. Unfortunately, she suddenly died a few days later, due to a presumed pulmonary embolism. Based on our experience, ECMO may be useful to manage pneumonia due to H7N9 avian influenza and ARDS in pregnant women.
Sujet(s)
Oxygénation extracorporelle sur oxygénateur à membrane/méthodes , Grippe humaine/thérapie , Pneumopathie virale/thérapie , Complications infectieuses de la grossesse/thérapie , /thérapie , Adulte , Issue fatale , Femelle , Humains , Sous-type H7N9 du virus de la grippe A/isolement et purification , Grippe humaine/virologie , Pneumopathie virale/virologie , Grossesse , Complications infectieuses de la grossesse/virologie , /virologieRÉSUMÉ
BACKGROUND: Chronic persistent asthma is characterized by airway remodeling, in which epithelial-mesenchymal transition (EMT) may play a significant role. Dehydroepiandrosterone (DHEA), a steroid hormone and testosterone analog, is considered as an important immunomodulating hormone. However, its role in EMT remains unclear. We sought to investigate whether transforming growth factor-ß1 (TGF-ß1) stimulates human bronchial epithelial cells (16HBE-14o) to undergo EMT, and whether this transition can be abrogated by DHEA. METHODS: The 16HBE-14o cells were stimulated with 5 ng/ml TGF-ß1 for 3 days to induce EMT, with or without DHEA pretreatment, and assayed for epithelial or mesenchymal markers using Western Blot. The involvement of phosphoinositide 3-kinase (PI3K) -mediated signaling pathway was also evaluated, the epithelial cells were also incubated with pharmacological approaches (agonists and antagonists of Akt, LY294002 or IGF-1) or flutamide, the antagonist of androgen receptor. Results were analyzed using nonparametric statistical tests. RESULTS: Our data demonstrate that treatment of 16HBE-14o cells with TGF-ß1 for 3 days induced EMT as reflected by conversion to the spindle-like morphology, loss of E-cadherin, and acquisition of a-smooth muscle actin (a-SMA). Pretreatment of 16HBE-14o cells with DHEA preserved the epithelial-like morphology, restored the expression of E-cadherin, and abolished the activation of a-SMA, and this effect is a PI3K-dependent mechanism. CONCLUSION: Our results indicate that TGF-ß1 induces EMT in a PI3K-dependent manner in 16HBE-14o cells. DHEA inhibits the bronchial epithelial to mesenchymal transition via the inhibition of PI3K/Akt-dependent signal pathway stimulated by TGF-ß1. Therefore, DHEA may be a useful therapy for asthma.
Sujet(s)
Bronches/effets des médicaments et des substances chimiques , Déhydroépiandrostérone/pharmacologie , Transition épithélio-mésenchymateuse/effets des médicaments et des substances chimiques , Bronches/anatomopathologie , Cellules cultivées , Relation dose-effet des médicaments , Cellules épithéliales/effets des médicaments et des substances chimiques , Cellules épithéliales/anatomopathologie , Humains , Phosphatidylinositol 3-kinases/physiologie , Protéines proto-oncogènes c-akt/physiologie , Récepteurs aux androgènes/physiologie , Facteur de croissance transformant bêta-1/pharmacologieRÉSUMÉ
Previous studies have consistently demonstrated that dopamine D1-like receptor (D1-like-R) signalling is implicated in the pathogenesis of experimental autoimmune encephalomyelitis and type I diabetes. Given that allergic asthma shares certain disease aetiology similarities with autoimmune diseases, we conducted studies in OVA-induced mice aiming to address the impact of D1-like-R signalling on the pathogenesis of allergic asthma. It was noted that blockade of D1-like-R signalling provided protection for mice against OVA-induced acute asthma. Particularly, treatment of OVA-induced mice with SCH23390, a D1-like-R antagonist, significantly attenuated inflammatory infiltration in the airways along with repressed goblet cell hyperplasia and mucus production, as well as airway resistance. By contrast, administration of SKF83959, a D1-like-R agonist, displayed the opposite effect. Blockade of D1-like-R signalling impaired Th17 function, as manifested by a significant reduction of Th17 cells in the spleen and bronchoalveolar lavage fluid. Mechanistic studies revealed that D1-like-R signalling enhances B-cell activating transcription factor activity, which then transcribes the expression of RORγt, a Th17 transcription factor; accordingly, D1-like-R signalling regulates Th17 differentiation to promote the development of allergic asthma. Taken together, the data obtained in the present suggest that blockade of D1-like-R signalling could be an effective therapeutic strategy for the prevention and treatment of allergic asthma in clinical practice.
Sujet(s)
Asthme/prévention et contrôle , Facteurs de transcription à motif basique et à glissière à leucines/antagonistes et inhibiteurs , Benzazépines/pharmacologie , Ovalbumine/toxicité , Récepteur dopamine D1/antagonistes et inhibiteurs , Cellules Th17/effets des médicaments et des substances chimiques , Maladie aigüe , Animaux , Asthme/induit chimiquement , Asthme/métabolisme , Lymphocytes B/effets des médicaments et des substances chimiques , Lymphocytes B/métabolisme , Lymphocytes B/anatomopathologie , Facteurs de transcription à motif basique et à glissière à leucines/génétique , Facteurs de transcription à motif basique et à glissière à leucines/métabolisme , Technique de Western , Liquide de lavage bronchoalvéolaire , Différenciation cellulaire/effets des médicaments et des substances chimiques , Prolifération cellulaire/effets des médicaments et des substances chimiques , Dopamine/métabolisme , Test ELISA , Femelle , Cytométrie en flux , Techniques immunoenzymatiques , Poumon/cytologie , Poumon/effets des médicaments et des substances chimiques , Poumon/métabolisme , Souris , Souris de lignée BALB C , Petit ARN interférent/génétique , Cellules Th17/cytologie , Cellules Th17/métabolismeRÉSUMÉ
Severe asthma is a chronic airway disease characterized by the Th2/Th17-polarized inflammation along with permanent airway remodeling. Despite past extensive studies, the exact role for Th2 and Th17 cytokines in asthmatic pathoetiology, particularly in the pathogenesis of bronchial epithelial-mesenchymal transition (EMT), is yet to be fully addressed. We herein conducted studies in 16-HBE cells and demonstrated that Th2-derived IL-4 and Th17-derived IL-17A provide a chronic inflammatory milieu that favors TGF-ß1 to induce bronchial EMT. A synergic action was noted between TGF-ß1, IL-4 and IL-17A in terms of induction of EMT. IL-4 and IL-17A synergized with TGF-ß1 to induce epithelial cells re-entering cell cycle, and to promote epithelial to mesenchymal morphological transistion, and by which they enhanced the capacity of TGF-ß1 to suppress E-cadherin expression, and to induce a-SMA expression in epithelial cells. Mechanistic studies revealed that this synergic action is coordinated by the regulation of ERK1/2 activity. Our results not only provide a novel insight into the understanding of the mechanisms underlying airway remodeling in asthmatic condition, but also have the potential for developing more effective therapeutic strategies against severe asthmatics in clinical settings.
Sujet(s)
Remodelage des voies aériennes/physiologie , Transition épithélio-mésenchymateuse/physiologie , Interleukine-17/métabolisme , Interleukine-4/métabolisme , Facteur de croissance transformant bêta-1/métabolisme , Asthme/immunologie , Asthme/métabolisme , Asthme/anatomopathologie , Technique de Western , Bronches/immunologie , Bronches/métabolisme , Bronches/anatomopathologie , Lignée cellulaire , Test ELISA , Humains , Inflammation/immunologie , Inflammation/métabolisme , Interleukine-17/immunologie , Interleukine-4/immunologie , Réaction de polymérisation en chaine en temps réel , Cellules Th17/immunologie , Lymphocytes auxiliaires Th2/immunologie , Facteur de croissance transformant bêta-1/immunologieRÉSUMÉ
In recent years, accumulating evidence suggests that atrial natriuretic peptide (ANP), a hormone widely known as a result of its significant effects on the cardiovascular system mediated by natriuretic peptide receptor A (NPRA), may play a nonnegligible role in the regulation of immune responses. In this study, we firstly investigated whether ANP signaling could regulate the differentiation and capacity of Th17 cells and discovered ANP-dose (10(-8)-10(-6)M) dependently indeed suppressed the differentiation of Th17 cells along with the reduced IL-17 production by polarizing naïve CD4(+) T cells isolated from splenocytes to Th17 phenotype in vitro. Moreover, ANP primarily signals through NPRA and cGMP-dependent protein kinase (PKG) which could be antagonized when pretreated with either ANP/NPRA signaling antagonist or PKG inhibitor. In addition, we also found that ANP signaling could upregulate the levels of phosphorylation of Akt which was hypothesized to be implicated in ANP-induced inhibition of Th17 development in our studies, and the effect of ANP on the development of murine Th17 cells seemed to be partially reversed when an inhibitor of phosphatidylinositol 3'-kinase (PI3K)/Akt had been performed in advance. Briefly, we showed for the first time that ANP signaling could suppress murine Th17 cell development from naïve CD4(+) T cells in vitro through NPRA/PKG pathway and the PI3K-Akt signal was implicated in the ANP-mediated suppression of Th17 development.
Sujet(s)
Cyclic GMP-Dependent Protein Kinases/génétique , Phosphatidylinositol 3-kinases/génétique , Protéines proto-oncogènes c-akt/génétique , Cellules Th17/effets des médicaments et des substances chimiques , Animaux , Facteur atrial natriurétique/immunologie , Facteur atrial natriurétique/pharmacologie , Différenciation cellulaire , Cyclic GMP-Dependent Protein Kinases/antagonistes et inhibiteurs , Cyclic GMP-Dependent Protein Kinases/immunologie , Régulation de l'expression des gènes/effets des médicaments et des substances chimiques , Interleukine-17/biosynthèse , Interleukine-17/immunologie , Souris , Phosphatidylinositol 3-kinases/immunologie , Inhibiteurs des phosphoinositide-3 kinases , Phosphorylation , Inhibiteurs de protéines kinases/pharmacologie , Protéines proto-oncogènes c-akt/antagonistes et inhibiteurs , Protéines proto-oncogènes c-akt/immunologie , Récepteur facteur natriurétique auriculaire/antagonistes et inhibiteurs , Récepteur facteur natriurétique auriculaire/génétique , Récepteur facteur natriurétique auriculaire/immunologie , Transduction du signal/effets des médicaments et des substances chimiques , Cellules Th17/cytologie , Cellules Th17/immunologieRÉSUMÉ
In traditional Chinese medicine, arsenous compounds, including arsenic trioxide (ATO), are often used to treat many diseases, which are safe and effective. Recently, studies have indicated that Th17- IL-17 involved in the pathogenesis and development of asthma. The goal of this study was to investigate the effect and mechanism of ATO on asthma, especially the Th17- IL-17 axis.We used oval bumin (OVA)-immunized mice as a model for asthma and treated mice with ATO or dexamethasone. The mice were then monitored airway responsiveness, airway inflammation, mucus production, IL-17 levels in BALF and the positive rate of Th17 cells. In vitro, CD4+ T cells from splenic cell suspensions were separated and purified. We measured the expression of IL-17 and caspase-12 protein in purified CD4+ T cells, and detected IL-17 levels in CD4+ T lymphocyte culture solution with or without ATO. Moreover, apoptosis, mitochondrial membrane potential, cytosolic calcium were analyzed. We found that ATO could reduce airway responsiveness, airway inflammation, mucus hyperplasia, the expression of IL-17 in BALF and the positive rate of Th17 cells at a level comparable to treatment with DXM. In vitro data suggested that ATO can induce CD4+ T cells apoptosis, cause mitochondrial dysfunction, Ca2+ overload and promote caspase-12 activation. Our study suggested that ATO had potential medical value for the treatment of human asthma..
Sujet(s)
Antiasthmatiques/pharmacologie , Composés de l'arsenic/pharmacologie , Asthme/traitement médicamenteux , Hyperréactivité bronchique/traitement médicamenteux , Interleukine-17/métabolisme , Poumon/effets des médicaments et des substances chimiques , Oxydes/pharmacologie , Cellules Th17/effets des médicaments et des substances chimiques , Animaux , Apoptose/effets des médicaments et des substances chimiques , Trioxyde d'arsenic , Asthme/immunologie , Asthme/métabolisme , Asthme/anatomopathologie , Asthme/physiopathologie , Hyperréactivité bronchique/immunologie , Hyperréactivité bronchique/métabolisme , Hyperréactivité bronchique/anatomopathologie , Hyperréactivité bronchique/physiopathologie , Tests de provocation bronchique , Calcium/métabolisme , Caspase-12/métabolisme , Cellules cultivées , Dexaméthasone , Modèles animaux de maladie humaine , Femelle , Poumon/immunologie , Poumon/métabolisme , Poumon/anatomopathologie , Poumon/physiopathologie , Potentiel de membrane mitochondriale/effets des médicaments et des substances chimiques , Souris , Souris de lignée BALB C , Mucus/métabolisme , Ovalbumine , Transduction du signal/effets des médicaments et des substances chimiques , Cellules Th17/immunologie , Cellules Th17/métabolisme , Cellules Th17/anatomopathologieRÉSUMÉ
Allergic asthma is a complex and chronic inflammatory airway disease. Interleukin-17 is a pro-inflammatory cytokine which plays critical role in the pathogenesis of allergic asthma. It has been reported that ß-arrestin2 regulated the development of allergic asthma at a proximal step in the inflammatory cascade. In this study, the influence of ß-arrestin2 on Interleukin-17 production and expression of CD4+ T lymphocytes in a murine asthma model was investigated. Splenic CD4+ T lymphocytes from wild-type mice and those from a murine asthma model were purified. CD4+ T lymphocytes from a murine asthma model were transfected with siRNAs targeting the ß-arrestin2 or were pretreated with the ERK1/2 inhibitor, PD98059. After stimulation, the protein expression of ß-arrestin2ãphosphorylated-ERK1/2 and IL-17 were detection by Western blot; the mRNA expression of IL-17 were detected by real-time PCR; the accumulation of IL-17 in supernatants were detected by ELISA. We found that ß-arrestin2ãphosphorylated-ERK1/2 and IL-17 expression in CD4+ T lymphocytes from a murine asthma model were increased compared with those from wild-type mice (p < 0.01). Treatment of CD4+ T lymphocytes with siRNAs targeting the ß-arrestin2 down-regulated phosphorylated- ERK 1/2 and IL-17 expression (p < 0.01). PD98059 decreased IL-17 production and expression in CD4+ T lymphocytes in a murine asthma model (p < 0.05). We conclude that ß-arrestin2 stimulated IL-17 production and expression of CD4+ T lymphocytes in a murine asthma model. The effect was partly mediated by ERK 1/2 activation. Targeting ß-arrestin2 biological activity could be a valid therapeutic approach for the treatment of allergic asthma.
Sujet(s)
Arrestines/immunologie , Asthme/immunologie , Lymphocytes T CD4+/immunologie , Interleukine-17/immunologie , Animaux , Arrestines/métabolisme , Asthme/métabolisme , Technique de Western , Lymphocytes T CD4+/métabolisme , Séparation cellulaire , Modèles animaux de maladie humaine , Test ELISA , Cytométrie en flux , Hypersensibilité/immunologie , Hypersensibilité/métabolisme , Interleukine-17/métabolisme , Activation des lymphocytes/immunologie , Système de signalisation des MAP kinases/immunologie , Souris , Souris de lignée BALB C , Mitogen-Activated Protein Kinase 1/immunologie , Mitogen-Activated Protein Kinase 1/métabolisme , Mitogen-Activated Protein Kinase 3/immunologie , Mitogen-Activated Protein Kinase 3/métabolisme , Interférence par ARN , Réaction de polymérisation en chaine en temps réel , bêta-ArrestinesRÉSUMÉ
Beta-arrestin 2 has been shown to participate in the pathogenesis of asthma by inducing Th2 cell migration to the lungs. Whether beta-arrestin 2 regulates cytokine production of CD4+ T cells is still unknown. The aim of the present study was to investigate the effect of beta-arrestin 2 on the cytokine production of CD4+ T lymphocytes and the mechanism involved in a mouse model for asthma. After silencing beta-arrestin 2 expression in CD4+ T lymphocytes from asthmatic mice by RNA interference (RNAi), the interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) levels in CD4+ T lymphocyte culture supernatants with or without terbutaline stimulation were determined. Cell-surface beta2 adrenergic receptor (beta2AR) as well as GATA3 expression of CD4+ T lymphocytes were also measured. CD4+ T lymphocytes of mice with allergic asthma expressed higher levels of beta-arrestin 2 on both mRNA and protein levels. beta-arrestin 2 RNAi decreased IL-4 (43.16%) and GATA3 (protein 77.21%, mRNA 62.98%) expression after terbutaline stimulation. Cell-surface beta2AR of CD4+ T lymphocytes decreased (15.27%) after terbutaline treatment, but recovered after beta-arrestin 2 RNAi down-modulation. These findings demonstrate that beta-arrestin 2 regulates IL-4 production and GATA3 expression of CD4+ T lymphocytes partly through the beta2AR signaling pathway in an allergic asthma model.