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INTRODUCTION: Apoptosis and chronic inflammation are the main phenotypes in chronic obstructive pulmonary disease (COPD) pathogenesis. Cigarette smoke exposure is the leading risk factor for COPD, which causes aberrant airway epithelial structure and function. As a non-classical calpain, the molecular function of calpain5 (CAPN5) in COPD remains unclear. This study investigated the role of CAPN5 in mediating cigarette smoke extract (CSE)-induced apoptosis and inflammation. METHODS: Immunohistochemistry (IHC) and Western blotting (WB) were performed to detect the location and expression of CAPN5. In vitro, BEAS-2B cells were transfected with CAPN5 siRNA or CAPN5 plasmid, followed by phosphate-buffered saline (PBS) or cigarette smoke extract (CSE) treatment. The protein expression levels of CAPN5, NF-κB p65, p-p65, IκBα, p-IκBα and apoptosis proteins (BCL-2, BAX) were measured by WB. Flow cytometry (FCM) was performed to analyze the cell apoptosis index. RESULTS: CAPN5 was mainly expressed in the airway epithelium and significantly decreased in the COPD-smoker and emphysema-mouse groups. Silencing CAPN5 significantly decreased the protein expression of BCL-2, IκBα, and increased p-p65 and BAX protein expression. Additionally, an increased apoptosis index was detected after silencing CAPN5. Moreover, overexpression of CAPN5 partly inhibited IκBα degradation and p65 activation, and reduced CSE-induced inflammation and apoptosis. CONCLUSIONS: These combined results indicate that CAPN5 could protect against CSE-induced apoptosis and inflammation, which may provide a potential therapeutic target for smoking-related COPD.
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Chronic obstructive pulmonary disease (COPD) is a significant global cause of morbidity and mortality currently. Long-term exposure of cigarette smoke (CS) inducing persistent inflammation, small airway remodeling and emphysematous lung are the distinguishing features of COPD. Ferroptosis, occurred in lung epithelial cells has recently been reported to be associated with COPD pathogenesis. DNA dioxygenase ten-eleven translocation 2 (TET2) is an important demethylase and its genetic mutation is associated with low forced expiratory volume in 1 s (FEV1) of lung function. However, its role in COPD remains elusive. Here, we found that TET2 regulates CS induced lipid peroxidation through demethylating glutathione peroxidase 4 (GPx4), thus alleviating airway epithelial cell ferroptosis in COPD. TET2 protein levels were mainly reduced in the airway epithelia of COPD patients, mouse models, and CS extract-treated bronchial epithelial cells. The deletion of TET2 triggered ferroptosis and further exaggerated CS-induced airway remodeling, inflammation, and emphysema in vivo. Moreover, we demonstrated that TET2 silencing intensified ferroptosis, while TET2 overexpression inhibited ferroptosis in airway epithelial cell treated with CSE. Mechanically, TET2 protected airway epithelial cells from CS-induced lipid peroxidation and ferroptosis through demethylating the promoter of glutathione peroxidase 4 (GPx4). Finally, co-administration of methylation inhibitor 5'-aza-2'-deoxycytidine (5-AZA) and the antioxidant N-acetyl-cysteine (NAC) have more protective effects on CS-induced COPD than either administration alone. Overall, our study reveals that TET2 is an essential modulator in the lipid peroxidation and ferroptosis of airway epithelial cell, and could act as a potential therapeutic target for CS-induced COPD.
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Fumar Cigarros , Dioxigenases , Ferroptose , Doença Pulmonar Obstrutiva Crônica , Camundongos , Animais , Humanos , Ferroptose/genética , Fumar Cigarros/efeitos adversos , Fosfolipídeo Hidroperóxido Glutationa Peroxidase/metabolismo , Pulmão/metabolismo , Doença Pulmonar Obstrutiva Crônica/genética , Células Epiteliais/metabolismo , Inflamação/metabolismo , DNA/metabolismo , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/metabolismo , Dioxigenases/metabolismo , Dioxigenases/farmacologiaRESUMO
Ovarian tumor family deubiquitinase 4 (OTUD4), a member of the OTU deubiquitinating enzyme, is implicated to decrease in cancer to regulate cell apoptosis. However, the role of OTUD4 in cigarette smoke induced epithelial cell apoptosis and its mechanism have not been elucidated. In this study, we showed that OTUD4 protein reduced in CSE treated mice and airway epithelial cells. OTUD4 silence aggravated cell apoptosis and emphysematous change in the lung tissue of cigarette smoke extract (CSE) treated mice. Additionally, restoration of OTUD4 in the lung of mice alleviated CSE induced apoptosis and emphysematous morphology change. The effect of OTUD4 on cell apoptosis was also confirmed in vitro. Through protein profile screening, we identified that OTUD4 may interact with plasminogen activator inhibitor 1(PAI-1). We further confirmed that OTUD4 interacted with PAI-1 for de-ubiquitination and inhibiting CSE induced PAI-1 degradation. Furthermore, the protective role of OTUD4 in airway epithelial cells apoptosis was blocked by PAI-1 deactivation. Taken together, our data suggest that OTUD4 regulates cigarette smoke (CS)-triggered airway epithelial cell apoptosis via modulating PAI-1 degradation. Targeting OUTD4/PAI-1 signaling might potentially provide a therapeutic target against the lung cell apoptosis in cigarette smoke (CS)-induced emphysema.
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Neoplasias Ovarianas , Inibidor 1 de Ativador de Plasminogênio , Animais , Camundongos , Feminino , Humanos , Inibidor 1 de Ativador de Plasminogênio/genética , Apoptose , Células Epiteliais , Pulmão , Proteases Específicas de UbiquitinaRESUMO
Background: Surgical and medical treatments are applied to pulmonary cryptococcosis (PC) in the real world, while the prognosis of different therapies is uncertain. This study investigated diagnosis, real-world therapy, follow-up outcomes, and prognosis factors, aiming to deepen our understanding of PC. Methods: Patients pathologically diagnosed with PC were retrospectively reviewed and followed up. Further comparisons and subgroup analyses were conducted in surgical and nonsurgical treatment individuals. Univariable and multivariable logistic regression methods were used to explore the risk factors associated with treatment failure. Results: One hundred and sixty-three patients were included in this study, of whom 92 underwent surgical removal of VATS or open lung surgery (68 of them received postoperative antifungal treatment) and 71 got antifungal drugs only. Compared with nonsurgical patients, surgical patients were more immunocompetent (73 [79.3%] cases vs 33 [46.5%]), showed milder symptoms and more limited pulmonary lesions. Although they had instant treatment response owing to lesions resection, there is no significant advantage in the rate of treatment failure. Multivariable regression showed independent predictive factors associated with treatment failure were polymorphonuclear (PMN)>6.30*109/L, albumin (Alb) <40g/L and antifungal dosage <400mg/d. Further analysis among patients with different immune statuses or symptoms demonstrated that sufficient antifungal dosage could reduce the rate of treatment failure. Conclusion: PC showed variable and nonspecific clinical features. PC patients with limited nodules/masses and mild symptoms often led to misdiagnosis and unnecessary lung resections. The potential risk factors including higher PMN and hypoalbuminemia could help clinicians to identify PC patients with poor treatment efficiency at an early stage. To note, sufficient antifungal dosage may improve the treatment outcomes.
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INTRODUCTION: Apoptosis of lung structural cells is a significant upstream event involved in COPD pathogenesis. This study was designed to explore whether pirfenidone (PFD) was able to attenuate apoptosis induced by cigarette smoke extract (CSE). METHODS: A method of intraperitoneal CSE injection to BALB/C mice was used to establish emphysema mouse model. Terminal deoxynucleotidyl transferase dUTPnick end labeling (TUNEL) assay was applied to evaluate apoptotic cell ratio in mouse lung tissue. The cell viability of HBECs exposed to different concentrations of PFD was measured by Cell Counting Kit-8 (CCK-8) assay. The apoptosis index (AI) of HBECs was tested by flow cytometry. Levels of apoptosis-related protein were determined by Western blotting. RESULTS: PFD treatment significantly decreased the AI value in emphysema mouse lung tissue by TUNEL. In HBECs, flow cytometry showed that PFD could significantly reduce AI led by CSE. Both in vitro and in vivo, protein levels of Bax and Cleaved-caspase 3 in CSE group significantly increased in contrast with the control group; while Bcl-2 protein level in CSE group was significantly decreased; moreover, PFD significantly reversed protein level changes of Bcl-2, Bax, and Cleaved-caspase 3 led by CSE. CONCLUSIONS: This study reveals that PFD may potentially protect against CSE induced apoptosis.
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BACKGROUND: Dihydroquercetin (DHQ) is a flavonoid with strong anti-inflammatory and antioxidant effects. However, its protective activity against cigarette smoke-induced ferroptosis in the pathogenesis of chronic obstructive pulmonary disease and its underlying mechanisms remain unclear. PURPOSE: The present study was conducted to investigate the protective role of DHQ in the pathogenesis of COPD in vivo and in vitro. METHODS: A cigarette smoke-induced COPD mouse model was established by cigarette smoke (CS) exposure combined with intraperitoneal injection of cigarette smoke extract (CSE). During the modeling process, the mice were intraperitoneally injected with DHQ daily. HBE cells were cultured with CSE with or without pretreatment with DHQ (40, 80 µM) or ML385 (10 µM). Cell viability was assessed by a cell counting kit 8 (CCK-8). The contents of malondialdehyde (MDA) and superoxide dismutase (SOD) were determined by MDA and SOD assay kits, respectively, and reactive oxygen species (ROS) generation was detected by DCFH-DA assays. Protein expression levels of solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPx4) and nuclear factor erythroid 2-related factor 2 (Nrf2) were measured by western blot. Lipid peroxidation was determined by C11-BODIPY staining. Transmission electron microscopy was used to observe the morphological features of the mitochondria. RESULTS: Treatment with DHQ significantly elevated ferroptosis-related protein (SLC7A11 and GPx4) expression in vivo and in vitro. The mRNA levels of SLC7A11 and GPx4 were also increased after DHQ treatment. The excessive MDA and ROS production and depleted SOD activity induced by CSE were reversed by DHQ. DHQ notably reduced the increased lipid peroxidation induced by CSE in HBE cells. In addition, treatment with DHQ attenuated the morphological changes in the mitochondria caused by CSE. Moreover, we also found that DHQ increased the levels of Nrf2 in a concentration-dependent manner in the cigarette smoke-induced COPD mouse model and CSE-treated HBE cells. Additionally, after administering an Nrf2-specific inhibitor, ML385, to HBE cells, the elevated SLC7A11 and GPx4 mRNA and protein levels induced by DHQ were reversed. Moreover, ML385 treatment attenuated the protective effect of DHQ on lipid peroxidation. CONCLUSION: Our results show that treatment with DHQ significantly reverses the ferroptosis induced by cigarette smoke both in vivo and in vitro via a Nrf2-dependent signaling pathway.
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Ferroptose , Doença Pulmonar Obstrutiva Crônica , Animais , Camundongos , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Quercetina/análogos & derivados , Fumaça/efeitos adversos , FumarRESUMO
BACKGROUND: Antioxidant and anti-inflammatory effects are two main pharmacological mechanisms of pirfenidone (PFD) besides the anti-fibrotic effect. This study aims to investigate whether PFD could mediate cigarette smoke extract (CSE) induced inflammation and oxidative stress in vitro and in vivo. METHODS: BALB/C mice and alveolar epithelial (A549) cells treated with CSE were established as disease models in vivo and in vitro. Effects of PFD treatment on disease models were further measured. Hematoxylin and eosin (HE) staining was used to evaluate the pathological changes in lung tissues of mice. CCK-8 assay kit was applied to measure the viability of A549 cells treated by different concentrations of PFD. Inflammation cytokine expression in cell supernatants was measured with ELISA kits. The mRNA and protein levels of inflammation and oxidative stress-related factors were determined by real-time quantitative polymerase chain reaction analysis (RT-qPCR) and Western blotting. Furthermore, myeloperoxidase (MPO), malondialdehyde (MDA), and total antioxidant capacity (T-AOC) were measured to detect the antioxidative activity of lung tissues. Moreover, an assay kit with fluorescent probe 2',7'-dichlorofluorescin diacetate (DCFH-DA) was used to evaluate the intracellular reactive oxygen species (ROS) generation. RESULTS: In vitro and in vivo, PFD significantly reversed TNF-α, IL-6, CCL2, SOD1, and CAT mRNA level changes led by CSE; in addition, PFD significantly decreased the ratios of p-p65 to p65, p-ikBα to ikBα and increased Nrf-2 protein level compared with CSE group. In mice, high-dose (100 mg/kg/d) PFD significantly reversed MPO and MDA increases induced by CSE. However, PFD didn't significantly reverse T-AOC decrease induced by CSE. In A549 cell supernatant, PFD dramatically reversed the elevated levels of TNF-α and IL-1ß induced by CSE. Furthermore, PFD could significantly reverse the increased level of ROS induced by CSE in A549 cells. CONCLUSION: Our study reveals the potential role of PFD in regulating inflammatory response and oxidative stress induced by CSE.
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Anti-Inflamatórios/uso terapêutico , Fumar Cigarros/efeitos adversos , Inflamação/tratamento farmacológico , Pulmão/patologia , Piridonas/uso terapêutico , Mucosa Respiratória/efeitos dos fármacos , Células A549 , Animais , Citocinas/metabolismo , Humanos , Inflamação/induzido quimicamente , Mediadores da Inflamação/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo , Extratos Vegetais/administração & dosagem , Espécies Reativas de Oxigênio/metabolismo , Mucosa Respiratória/fisiologia , Transdução de SinaisRESUMO
BACKGROUND: This study aimed to evaluate the C-reactive protein to serum albumin ratio (CAR) to predict prognosis in COPD patients with acute exacerbations (AECOPD). METHODS: A retrospective cohort study of AECOPD patients, admitted to a large tertiary hospital between January 2017 and June 2018, was conducted. Univariate and multivariate logistic regression models were built to assess the relationship between variables and different clinical outcomes in one-year follow up. In addition, Kaplan-Meier method was used to estimate the relationship between CAR and the time to first rehospitalization due to acute exacerbation of COPD. RESULTS: A total of 167 AECOPD patients were included in this study, with an overall age of 68.5 ± 9.4 years. Both univariate and multivariate logistic regression analysis demonstrated that CAR at admission was significantly associated with rehospitalization and frequent exacerbations in COPD patients (p < 0.05). Kaplan-Meier curve showed that the rehospitalization event-free rate was signiï¬cantly higher in the low CAR group than the high CAR group (p < 0.01). CONCLUSIONS: As an easily available parameter, CAR at admission can be an independent predictor for rehospitalization and frequent exacerbations in COPD patients with acute exacerbations.
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Proteína C-Reativa , Doença Pulmonar Obstrutiva Crônica , Idoso , Biomarcadores , Progressão da Doença , Humanos , Pessoa de Meia-Idade , Prognóstico , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Estudos Retrospectivos , Albumina SéricaRESUMO
OBJECTIVE: To determine if there is a relationship between the levels of serum uric acid and the different Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages (1-4) classified by the severity of the airflow limitation in patients with stable chronic obstructive pulmonary disease (COPD). METHODS: Electronic databases, including PubMed®, Embase®, Web of Science™ and China National Knowledge Infrastructure (CNKI), were searched from inception to December 2018. Observational studies that reported serum uric acid levels in stable COPD patients were included. Two investigators independently extracted data and RevMan version 5.3 was used to carry out the statistical analyses. RESULTS: Seven studies with 932 stable COPD patients and 401 healthy control subjects were included in this meta-analysis. Serum uric acid levels were significantly higher in stable COPD patients compared with healthy control subjects (mean difference [MD] 1.91, 95% confidence interval [CI] 1.55, 2.28). Serum uric acid levels were significantly lower in the GOLD 1+2 subgroup compared with the GOLD 3+4 subgroup (MD -1.39, 95% CI -1.63, -1.15). CONCLUSION: Serum uric acid might be a useful biomarker for identifying disease severity in stable COPD patients, but further studies are needed to confirm this finding.
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Doença Pulmonar Obstrutiva Crônica , Ácido Úrico , Biomarcadores , China , Humanos , Pulmão , Doença Pulmonar Obstrutiva Crônica/diagnósticoRESUMO
Obstructive sleep apnea syndrome (OSAS), a state of sleep disorder, is characterized by repetitive apnea, chronic hypoxia, oxygen desaturation, and hypercapnia. Previous studies have revealed that intermittent hypoxia (IH) conditions in OSAS patients elicited neuron injury (especially in the hippocampus and cortex), leading to cognitive dysfunction, a significant and extraordinary complication of OSAS patients. The repeated courses of airway collapse and obstruction in OSAS patients resulted in apnea and arousal during sleep, leading to IH and excessive daytime sleepiness (EDS) and subsequently contributing to the development of inflammation. IH-mediated inflammation could further trigger various types of cognitive dysfunction. Many researchers have found that, besides continuous positive airway pressure (CPAP) treatment and surgery, anti-inflammatory substances might alleviate IH-induced neurocognitive dysfunction. Clarifying the role of inflammation in IH-mediated cognitive impairment is crucial for potentially valuable therapies and future research in the related domain. The objective of this article was to critically review the relationship between inflammation and cognitive deficits in OSAS.
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Disfunção Cognitiva/patologia , Inflamação/patologia , Apneia Obstrutiva do Sono/patologia , Córtex Cerebral/patologia , Disfunção Cognitiva/etiologia , Hipocampo/patologia , Humanos , Microglia/patologia , Apneia Obstrutiva do Sono/complicaçõesRESUMO
BACKGROUND: China announced the implementation of the universal two-child policy in Oct, 2015; every couple was allowed to have two children. However, its influences on maternal well-being and infants' outcomes are still to be discovered. OBJECTIVES: To detect influences of the universal two-child policy. To provide information for maternal health care under the new policy. STUDY DESIGN: This study enrolled 859 and 1230 women who delivered their second child (hereafter second-time mothers) before and after the policy's implementation, respectively, and the data included maternal demographic characteristics, gestational complications, delivery mode and infants' outcomes. RESULTS: After the policy's implementation, the proportion of second-time mothers with advanced age increased significantly. The advanced gestational age is well acknowledged to correlate with higher risk during the pregnancy, both for pregnant women and their babies. However, in our study, the incidence of hypertensive disorders in pregnancy, placenta previa and postpartum haemorrhage decreased significantly after the introduction of the policy and no differences were noted in other gestational complications. Moreover, the hospitalization time was shortened, and caesarean delivery was chosen less frequently. As for the infants, foetal distress exhibited an alleviation and the incidence of premature labour and low birth weight decreased as well. CONCLUSIONS: Even though the age of second-time mothers increased after the introduction of the universal two-child policy, their general gestational health condition improved and their infants also exhibited a better outcome, which might be attributed to the improvement of China's maternal medical care system.