The impact of claudin 5 on clinical variables in patients with asthma

이푸른하늘,김병곤,서현정,백애린,박종숙,이준혁,박성우,김도진,박춘식,장안수 대한결핵 및 호흡기학회 2015 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.120 No.-

Background: The tight junction (TJ) protein,claudin 5 (CLDN5) is critical to the control of endothelial cellular polarity and pericellular permeability. The role of CLDN5 in asthma remains uncovered. Objective: The aim of this study was to determine CLDN5 levels in patients with bronchial asthma and to evaluate relationship between CLDN5 levels with clinical variables. Methods: Fifty patients with bronchial asthma (mean age: 54.9±14.1 years, mean disease duration: 7.28±13ㆍ5 years) and 25 healthy controls were enrolled. CLDN5 levels were checked in stable and exacerbated state of disease of asthmatic patients. Results: The mean plasmaCLDN5 level of patients with bronchial asthma was 0.665 ±0.10 ng/mg and 0.955 ±0.05 ng/mg in healthy controls (p=0.023). The mean plasma CLDN5 levelswas 0.930 ± 0.095 ng/mg in exacerbatedstate of asthmatics and 0.399 ± 0.02 ng/mg in controlstate of those (p=0.001). The plasmaCLDN5levelswere correlated with eosinophils (r=-0ㆍ237, P=0ㆍ019), total IgE (r=-0ㆍ225, P=0ㆍ012), FEV1 % pred.(r=-0ㆍ415, P=0ㆍ001), and FEV1/FVC (r=-0ㆍ294, P=0ㆍ004). The plasmaCLDN5levelswerenot correlated with sex, PC20 methacholine, smoke amount, and body mass index. Steroid treatment with stable patients reduced plasma CLDN5 levels (0.330 ± 0.028 ng/ml vs. 0.426 ± 0.035 ng/ml, p=0.03) Conclusion: CLDN5 may bea marker for asthma exacerbation and implicated in the pathogenesis of bronchial asthma.

The changes of lung microbiota in a mouse model exposed to air pollutants

이푸른하늘,김병곤,홍지수,백애린,이준혁,박성우,김도진,장안수 대한결핵 및 호흡기학회 2018 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.126 No.-

Background: Environmental microbes have been associated with both protective and adverse lung effects in chronic lung disease. Although the gut microbiome for human health was relatively well studied, the respiratory microbiome’s role in the human response to inhaled pollutants is largely unknown. Objectives: Our aim was to characterize the bacterial microbiome in lung samples collected in a mouse model exposed to nanoparticles. Methods: Mice were exposed to TiO2 particles (200 μg/m3 for 1hr, and saline control on days 1-5, respectively n=8) using an ultrasonic nebulizer. We determined the composition of microbial communities present in lung samples by amplifying and sequencing regions of rRNA genes from bacteria (16S). Sequencing of amplified 16S regions was performed on the Roche-454 Life Sciences Titanium pyrosequencing platform. Also airway inflammation and responsiveness were correlated with lung microbiota. Results: Relative to controls, TiO2 exposure mice revealed increased hyperresponsiveness and inflammatory cells. Pseudomonas, Acinetobacter, Brucella, Mesorhizobium, Enhydrobacter, Methylobacterium, Rhizobium, Chryseobacterium, Brevundimonas, Deinococcus, and Micrococcus were differently observed between TiO2 exposure and control miceta. One of them, Deinococcus significantly increased in TiO2 exposure mice compared to control mice. The airway inflammatory cells correlated with Deinococcus. Conclusion: Nanoparticles changed lung microbial taxa, suggesting that exposure to air pollutants may cause to lung inflammation via alterations in the lung microbiota.

N-acetylcysteine decreases airway inflammation and responsiveness in asthma by modulating claudin 18 expression

이푸른하늘,Jisu Hong,An-Soo Jang 대한내과학회 2020 The Korean Journal of Internal Medicine Vol.35 No.5

Background/Aims: N-acetylcysteine (NAC) affects signaling pathways involved in apoptosis, angiogenesis, cell growth and arrest, redox-regulated gene expression, and the inflammatory response. However, it is not known how the signal mechanism for tight junctional protein claudin (CLDN) 18 is regulated in asthma patients. Methods: To investigate the effects of NAC on CLDN18 expression in a mouse model of asthma, and to assess plasma levels of CLDN18 in asthma patients. A murine model of asthma induced by ovalbumin (OVA) was established using wild-type BALB/c female mice, and the levels of CLDNs, phosphorylated-pyruvate dehydrogenase kinase 1 (p-PDK1), and protein kinase B (Akt) pathway proteins following NAC treatment were examined by Western blotting and immunohistochemistry. In addition, the plasma levels of CLDN18 were evaluated in asthmatic patients and control subjects. Results: NAC diminished OVA-induced airway hyper-responsiveness and inflammation. Levels of CLDN18 protein were higher in lung tissue from OVA mice than tissue from control mice, and were increased by treatment with NAC or dexamethasone. Treatment with NAC or dexamethasone decreased the OVA-induced increase in interleukin-1α protein levels. Although treatment with NAC increased OVA-induced p-PDK1 protein levels, it decreased phosphorylated Akt (pAkt)/Akt levels. Soluble CLDN18 levels were lower in patients with asthma than in controls and were correlated with the percentage of neutrophils, forced expiratory volume in 1 second (FEV1)/forced vital capacity % (FVC%) and FEV1%. Conclusions: CLDN18 plays a role in the pathogenesis of asthma and NAC diminishes airway inflammation and responsiveness by modulating CLDN18 expression.

The Impact of Diesel Exhaust Particles on Tight Junctional Proteins on Nose and Lung in a Mouse Model

이푸른하늘,Kim Byeong-Gon,Park Moo-Kyun,Hong Jisu,Lee Yun-Gi,Jang An-Soo 대한천식알레르기학회 2021 Allergy, Asthma & Immunology Research Vol.13 No.2

P-42 Epithelial tight junction protein, claudin-4 involved in asthma pathogenesis

이푸른하늘,김병곤,이선혜,백애린,박종숙,이준혁,박성우,김도진,박춘식,장안수 대한결핵 및 호흡기학회 2016 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.121 No.-

Background: Claudins are tight junction proteins that regulate paracellular ion permeability of epithelium and endothelium. Claudin 4 has been reported to function as a paracellular sodium barrier and is one of three major claudins expressed in lung alveolar epithelial cells. But the possible role of CLDN4 in bronchial asthma has not yet been studied. In this study we aimed to elucidate the role of CLDN4 in the pathogenesis of bronchial asthma. Methods: Using mice sensitized with ovalbumin (OVA) and challenged with OVA, as well as mice sensitized and challenged with saline, we investigated whether CLDN4 is involved in the pathogenesis of bronchial asthma. Moreover, we also evaluated relationship between the levels of CLDN4 levels in blood from asthmatic patients and clinical variables. Results: The OVA-OVA mice showed that the transcript and protein of CLDN4 in lung tissue were significantly increased after OVA challenge, and significant increases in tight junction breaks and the density of CLDN4 staining. Moreover, we found that CLDN4 in blood from asthmatic patients were increased compared with those from healthy control subjects. Plasma CLDN4 levels were significantly high in exacerbated patients compared with control patients with BA. The plasma CLDN4 level was correlated with total IgE, FEV1 % pred, and FEV1/FVC. Conclusion: These results indicate that CLDN4 might be implicated in the pathogenesis of bronchial asthma.

Effect of TiO2 Nanoparticles on Inflammasome-Mediated Airway Inflammation and Responsiveness

김병곤,이푸른하늘,이선혜,박무균,장안수 대한천식알레르기학회 2017 Allergy, Asthma & Immunology Research Vol.9 No.3

Purpose: Nanoparticles (NPs) may cause cell and tissue damage, leading to local and systemic inflammatory responses and adverse effects on health due to the inhalation of particulate matter. The inflammasome is a major regulator of inflammation through its activation of pro-caspase-1, which cleaves pro-interleukin-1β (pro-IL-1β) into its mature form and may induce acute and chronic immune responses to NPs. However, little is known about the response of the inflammasome to NP exposure via the airways in asthma. The aim of this study was to identify the impact of titanium dioxide (TiO2) NPs on inflammasome in a mouse model of allergic asthma. Methods: Mice were treated with ovalbumin (OVA) or TiO2 NPs. IL- 1β, IL-18, NAIP, CIITA, HET-E, TP-2 (NACHT), leucine-rich repeat (LRR), pyrin domain-containing protein 3 (NLRP3), and caspase-1 were assessed by Western blotting. Caspase-1 was assessed by immunohistochemistry (IHC). Levels of reactive oxygen species (ROS)—as markers of oxidative damage— and the mediators 8-isoprostane and carbonyl were measured by enzyme-linked immunosorbent assay (ELISA). Results: Airway hyperresponsiveness (AHR) and inflammation were increased in OVA-sensitized/challenged mice, and these responses were exacerbated by exposure to TiO2 NPs. NP treatment increased IL-1β and IL-18 expression in OVA-sensitized/challenged mice. NPs augmented the expression of NLRP3 and caspase- 1, leading to production of active caspase-1 in the lung. Caspase-1 expression was increased and exacerbated by TiO2 NP exposure in OVAsensitized/ challenged mice. ROS levels tended to be increased in OVA-sensitized/challenged and OVA-sensitized/challenged-plus-TiO2 NP-exposed mice. Conclusions: Our data demonstrated that inflammasome activation occured in asthmatic lungs following NP exposure, suggesting that targeting the inflammasome may assist in controling NP-induced airway inflammation and hyperresponsiveness.

P-104 Association of Annexin A5 with lung inflammation and fibrosis in asthma

이선혜,이푸른하늘,김병곤,서현정,백애린,박종숙,이준혁,박성우,김도진,박춘식,장안수 대한결핵 및 호흡기학회 2017 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.124 No.-

Background: Annexin-A5 (ANXA5) have a calcium channel activity and a potential role in cellular signal transduction, inflammation, growth and differentiation, and associated with anticoagulatory, pro-apoptotic activities, fibrosis. However, the exact role of ANXA5 in asthma remains to be determined. Objective: The aim of this study was to identify the role of ANXA5 in bronchial asthma, and to know relationship between clinical variables and the blood level. Methods: Female BALB/c mice were sensitized and challenged with ovalbumin (OVA-OVA), saline (control) or titanium dioxide (TiO₂). We investigated ANXA5 levels using ELISA assays or immunoblotting and immunohistochemical staining. Lung fibrosis and transforming growth factor β1 (TGF-β1) was observed in the mouse lung using Western blots. Moreover, we also checked ANXA5 levels in blood from asthmatic patients (stable and exacerbated states). Results: The protein of ANXA5 in lung tissue decreased in OVA-OVA mice compared with control mice. Lung ANXA5 and TGF-β1 protein was higher increased in OVA plus TiO₂ mice than in OVA-OVA mice. Dermatophagoides pteronyssinus treatment increased in lung ANXA5 protein in human lung fibroblast cells, but decreased lung ANXA5 protein in human lung epithelial cells. Plasma ANXA5 levels were decreased in asthmatic patients compared with healthy controls, and significantly higher in exacerbated patients compared with stable patients with bronchial asthma. ANXA5 correlated with FEV₁%, FEV₁/FVC%. Conclusion: Our results suggest that ANXA5 has potential role in asthma pathogenesis.

Long-Term Effects of Diesel Exhaust Particles on Airway Inflammation and Remodeling in a Mouse Model

김병곤,이푸른하늘,이신화,김영은,신미용,강예나,배성환,김민정,임태연,박춘식,장안수 대한천식알레르기학회 2016 Allergy, Asthma & Immunology Research Vol.8 No.3

Purpose: Diesel exhaust particles (DEPs) can induce and trigger airway hyperresponsiveness (AHR) and inflammation. The aim of this study was to investigate the effect of long-term DEP exposure on AHR, inflammation, lung fibrosis, and goblet cell hyperplasia in a mouse model. Methods: BALB/c mice were exposed to DEPs 1 hour a day for 5 days a week for 3 months in a closed-system chamber attached to a ultrasonic nebulizer (low dose: 100 μg/m3 DEPs, high dose: 3 mg/m3 DEPs). The control group was exposed to saline. Enhanced pause was measured as an indicator of AHR. Animals were subjected to whole-body plethysmography and then sacrificed to determine the performance of bronchoalveolar lavage and histology. Results: AHR was higher in the DEP group than in the control group, and higher in the high-dose DEP than in the low-dose DEP groups at 4, 8, and 12 weeks. The numbers of neutrophils and lymphocytes were higher in the high-dose DEP group than in the low-dose DEP group and control group at 4, 8, and 12 weeks. The levels of interleukin (IL)-5, IL-13, and interferon-γ were higher in the low-dose DEP group than in the control group at 12 weeks. The level of IL-10 was higher in the high-dose DEP group than in the control group at 12 weeks. The level of vascular endothelial growth factor was higher in the low-dose and high-dose DEP groups than in the control group at 12 weeks. The level of IL-6 was higher in the low-dose DEP group than in the control group at 12 weeks. The level of transforming growth factor-β was higher in the high-dose DEP group than in the control group at 4, 8, and 12 weeks. The collagen content and lung fibrosis in lung tissue was higher in the high-dose DEP group at 8 and 12 weeks. Conclusions: These results suggest that long-term DEP exposure may increase AHR, inflammation, lung fibrosis, and goblet cell hyperplasia in a mouse model.

P-68 The impact of endothelial tight junction protein, claudin 5 on COPD

김병곤,이푸른하늘,이선혜,백애린,박종숙,이준혁,박성우,김도진,박춘식,장안수 대한결핵 및 호흡기학회 2016 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.121 No.-

Background: The tight junction protein, claudin 5 (CLDN5) is critical to the control of endothelial cellular polarity and pericellular permeability. The role of CLDN5 in COPD remains uncovered. Objective: The aim of this study was to check CLDN5 levels in patients with COPD and to know relationship between CLDN5 levels with clinical variables. Methods: Thirty patients with COPD (mean age: ± years) and 25 healthy controls were enrolled to the study. Plasma CLDN5 level was checked in stable and in exacerbated state of patients with COPD. Results: The mean plasma CLDN5 level of total patients with COPD was 0.675 ± 0.034 ng/ml and 6.918 ± 0.78 ng/ml in healthy controls (p=0.001). The mean plasma CLDN5 level was 0.713 ± 0.043 ng/ml in exacerbated and 0.638± 0.053 ng/ml in stable patients with COPD (p=0.001). The plasma CLDN5 level in stable patients of COPD was correlated with smoking amount (r=-0·603, P=0·001), initial WBC(r=-0·447, P=0·009), BMI (r=0·4, P=0·001), FEV1 % pred.(r=0·573, P=0·001), and FEV1/FVC (r=0·609, P=0·001). The plasma CLDN5 level was not correlated with age. Conclusion: CLDN5 may be a marker for COPD exacerbation and implicated in the pathogenesis of COPD.

Titanium dioxide nanoparticles induced inflammasome IL-1β and IL-18 airway hyperresponsiveness and inflammation in a mouse model of asthma

김병곤,이푸른하늘,서현정,백애린,박종숙,이준혁,박성우,김도진,박춘식,장안수 대한결핵 및 호흡기학회 2015 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.120 No.-

Background: Nanoparticles (NPs) may pose adverse health effects due to particulate matter inhalation and induces cell and tissue damage, causing local and systemic inflammatory responses. The inflammasome is a major regulator of inflammation through its activation of pro-caspase-1, which cleaves pro-IL-1β into its mature form and may signal acute and chronic immune responses to NPs. However, little is known about the inflammasome of NPs exposure via the airways in asthma. Results: Mice were treated with saline Sham, OVA, or TiO2 NPs. Lung IL-1β, IL-18, NACHT, LRR and PYD domains-containing protein 3 (NLRP3) and caspase-1 levels were assessed with Western Blot. Caspase-1 was checked by immunohistochemical staining. Reactive oxygen species (ROS) were measured for the marker 8-isoprostance by ELISA. Airway inflammation and hyperresponsiveness increased in OVA sensitized mice and these responses were exaggerated by TiO2 NPs exposure. TiO2 NPs treatment increased IL-1β and IL-18 protein expression in OVA sensitized mice. TiO2 NPs augmented the expression of NLRP3 and caspase-1 leading to the formation of an active caspase-1 in the lung. Lung caspase-1 expression was increased in OVA sensitized mice and these responses were exaggerated by TiO2 NPs exposure. ROS tended to be increased in OVA sensitized mice and in OVA sensitized plus TiO2 NPs exposed mice. Conclusion: Our data demonstrate that inflammasome activation occurs in asthmatic lungs following NPs exposure, suggesting that targeting the inflammasome may help control NPs-induced airway inflammation and responsiveness.