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1.
Protein & Cell ; (12): 717-733, 2021.
Article in English | WPRIM | ID: wpr-888715

ABSTRACT

The coronavirus disease 2019 (COVID-19) pandemic is caused by infection with the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which is spread primary via respiratory droplets and infects the lungs. Currently widely used cell lines and animals are unable to accurately mimic human physiological conditions because of the abnormal status of cell lines (transformed or cancer cells) and species differences between animals and humans. Organoids are stem cell-derived self-organized three-dimensional culture in vitro and model the physiological conditions of natural organs. Here we showed that SARS-CoV-2 infected and extensively replicated in human embryonic stem cells (hESCs)-derived lung organoids, including airway and alveolar organoids which covered the complete infection and spread route for SARS-CoV-2 within lungs. The infected cells were ciliated, club, and alveolar type 2 (AT2) cells, which were sequentially located from the proximal to the distal airway and terminal alveoli, respectively. Additionally, RNA-seq revealed early cell response to virus infection including an unexpected downregulation of the metabolic processes, especially lipid metabolism, in addition to the well-known upregulation of immune response. Further, Remdesivir and a human neutralizing antibody potently inhibited SARS-CoV-2 replication in lung organoids. Therefore, human lung organoids can serve as a pathophysiological model to investigate the underlying mechanism of SARS-CoV-2 infection and to discover and test therapeutic drugs for COVID-19.


Subject(s)
Adenosine Monophosphate/therapeutic use , Alanine/therapeutic use , Alveolar Epithelial Cells/virology , Antibodies, Neutralizing/therapeutic use , COVID-19/virology , Down-Regulation , Drug Discovery , Human Embryonic Stem Cells/metabolism , Humans , Immunity , Lipid Metabolism , Lung/virology , RNA, Viral/metabolism , SARS-CoV-2/physiology , Virus Replication/drug effects
2.
Article in Chinese | WPRIM | ID: wpr-878923

ABSTRACT

This study aimed to assess whether chrysin(ChR) can inhibit epithelial-mesenchymal transition(EMT) of type Ⅱ alveolar epithelial cell and produce anti-pulmonary fibrosis effect by regulating the NF-κB/Twist 1 signaling pathway. Sixty rats were randomly divided into the control group, the bleomycin(BLC) group, BLC+ChR(50 mg·kg~(-1)) group and BLC+ChR(100 mg·kg~(-1)) group, with 15 rats in each group. The pulmonary fibrosis model was induced by intratracheal injection of BLC(7 500 U·kg~(-1)). Rats were orally administered with different doses of ChR after BLC injection for 28 days. The cells were divided into control group, TGF-β1 group(5 ng·mL~(-1)), and TGF-β1+ChR(1, 10, 100 μmol·L~(-1)) groups. The type Ⅱ alveolar epithelial cells were treated with TGF-β1 for 24 h, and then treated with TGF-β1 for 48 h in the presence or absence of different doses of ChR(1, 10 and 100 μmol·L~(-1)). The morphological changes and collagen deposition in lung tissues were analyzed by HE staining, Masson staining and immunohistochemistry. The mRNA and protein expression levels of collagen Ⅰ, E-cadherin, zonula occludens-1(ZO-1), vimentin, alpha smooth muscle actin(α-SMA), inhibitor of nuclear factor kappa B alpha(IκBα), nuclear factor-kappa B p65(NF-κB p65), phospho-NF-κB p65(p-p65) and Twist 1 in lung tissues and cells were detected by qPCR and Western blot, respectively. The animal experiment results showed that as compared with the BLC group, after administration of ChR for 28 days, bleomycin-induced pulmonary fibrosis in rats was significantly relieved, collagen Ⅰ expression in lung tissues was significantly reduced(P<0.05 or P<0.01), and EMT of alveolar epithelial cells was obviously inhibited [the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], concomitantly with significantly reduced IκBα and p65 phosphorylation level in cytoplasm and decreased NF-κB p65 and Twist 1 expression in nucleus(P<0.05 or P<0.01). The cell experiment results showed that different doses of ChR(1, 10 and 100 μmol·L~(-1)) significantly reduced TGF-β1-induced collagen Ⅰ expression(P<0.05 or P<0.01), significantly inhibited EMT of type Ⅱ alveolar epithelial cells[the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], and inhibited IκBα and p65 phosphorylation in cytoplasm and down-regulated NF-κB p65 and Twist 1 expression in nucleus induced by TGF-β1(P<0.05 or P<0.01). The results suggest that ChR can reverse EMT of type Ⅱ alveolar epithelial cell and alleviate pulmonary fibrosis in rats, and its mechanism may be associated with reducing IκBα phosphorylation and inhibiting NF-κB p65 phosphorylation and nuclear transfer, thus down-regulating Twist 1 expression.


Subject(s)
Alveolar Epithelial Cells/metabolism , Animals , Epithelial-Mesenchymal Transition , Flavonoids , NF-kappa B/metabolism , Rats , Signal Transduction , Transforming Growth Factor beta1/genetics
3.
Chinese Medical Journal ; (24): 261-274, 2021.
Article in English | WPRIM | ID: wpr-878043

ABSTRACT

There have been recent extensive studies and rapid advancement on the pathogenesis underlying idiopathic pulmonary fibrosis (IPF), and intricate pathogenesis of IPF has been suggested. The purpose of this study was to clarify the logical relationship between these mechanisms. An extensive search was undertaken of the PubMed using the following keywords: "etiology," "pathogenesis," "alveolar epithelial cell (AEC)," "fibroblast," "lymphocyte," "macrophage," "epigenomics," "histone," acetylation," "methylation," "endoplasmic reticulum stress," "mitochondrial dysfunction," "telomerase," "proteases," "plasminogen," "epithelial-mesenchymal transition," "oxidative stress," "inflammation," "apoptosis," and "idiopathic pulmonary fibrosis." This search covered relevant research articles published up to April 30, 2020. Original articles, reviews, and other articles were searched and reviewed for content; 240 highly relevant studies were obtained after screening. IPF is likely the result of complex interactions between environmental, genetic, and epigenetic factors: environmental exposures affect epigenetic marks; epigenetic processes translate environmental exposures into the regulation of chromatin; epigenetic processes shape gene expression profiles; in turn, an individual's genetic background determines epigenetic marks; finally, these genetic and epigenetic factors act in concert to dysregulate gene expression in IPF lung tissue. The pathogenesis of IPF involves various imbalances including endoplasmic reticulum, telomere length homeostasis, mitochondrial dysfunction, oxidant/antioxidant imbalance, Th1/Th2 imbalance, M1-M2 polarization of macrophages, protease/antiprotease imbalance, and plasminogen activation/inhibition imbalance. These affect each other, promote each other, and ultimately promote AEC/fibroblast apoptosis imbalance directly or indirectly. Excessive AEC apoptosis and impaired apoptosis of fibroblasts contribute to fibrosis. IPF is likely the result of complex interactions between environmental, genetic, and epigenetic factors. The pathogenesis of IPF involves various imbalances centered on AEC/fibroblast apoptosis imbalance.


Subject(s)
Alveolar Epithelial Cells , Apoptosis , Endoplasmic Reticulum Stress , Fibroblasts , Humans , Idiopathic Pulmonary Fibrosis/genetics
4.
Rev. Ciênc. Méd. Biol. (Impr.) ; 19(4): 602-605, dez 30, 2020.
Article in English | LILACS | ID: biblio-1355219

ABSTRACT

Introduction: the severe acute respiratory syndrome ­ coronavirus 2 (SARS Cov-2), leads to a diffuse alveolar deterioration due infection of type II pneumocytes. The type II pneumocytes are involved in synthesis and secretion of pulmonary surfactant in pulmonary alveoli. Objective: the purpose of this study is to discuss the indication of surfactant replacement as a potential adjunctive treatment modality for SARS CoV-2, similarly treatment to neonatal respiratory distress syndrome. Methodology: we argue that SARS can be triggered by surfactant deficiency secondary to production deficiency determined by type 2 pneumocyte injuries. In this sense, we carried out a bibliographic review. Conclusion: thus, the replacement of human surfactant could be a potential treatment modality for SARS CoV-2, in the same way that it is indicated for the treatment of neonatal respiratory distress syndrome.


Introdução: a síndrome respiratória aguda grave ­ coronavírus 2 (SARS Cov-2), leva a uma deterioração alveolar difusa devido à infecção do pneumócitos tipo II. Os pneumócitos tipo II estão envolvidos na síntese e secreção de surfactante pulmonar nos alvéolos pulmonares. Objetivo: o objetivo deste estudo é discutir a indicação de reposição de surfactante como uma potencial modalidade de tratamento adjuvante para SARS CoV-2, similarmente ao tratamento da síndrome do desconforto respiratório neonatal. Metodologia: argumentamos que a SARS pode ser desencadeada pela deficiência de surfactante, secundária à deficiência da sua produção determinada por lesões de pneumócitos tipo 2. Nesse sentido, realizamos uma revisão bibliográfica. Conclusão: o uso de surfactante humana pode ser uma potencial modalidade de tratamento para a SARS CoV-2, da mesma forma que é indicada para o tratamento da síndrome do desconforto respiratório neonatal.


Subject(s)
Pulmonary Surfactants , Severe Acute Respiratory Syndrome , Alveolar Epithelial Cells , SARS-CoV-2 , Review , Annual Report
5.
Int. j. odontostomatol. (Print) ; 14(4): 501-507, dic. 2020. graf
Article in Spanish | LILACS | ID: biblio-1134527

ABSTRACT

RESUMEN: Un nuevo coronavirus (SARS-CoV-2) ha sido reconocido como el agente etiológico de una misteriosa neumonía originada en Wuhan, China. La OMS ha nombrado a la nueva enfermedad como COVID-19 y, además, la ha declarado pandemia. Taxonómicamente, SARS-CoV-2 pertenece al género de los betacoronavirus junto con SARS-CoV y MERS-CoV. SARS-CoV-2 utiliza la enzima convertidora de la angiotensina 2 (ACE2) como el receptor objetivo para el ingreso en una célula huésped. La expresión de ACE2 en células de tejidos humanos podría indicar un potencial riesgo de reconocimiento por parte del virus y, por ende, ser susceptibles a la infección. Mediante algunas técnicas de laboratorio y de bioinformática, se ha visto una alta presencia de ACE2 en células epiteliales alveolares tipo II de pulmón y en enterocitos del intestino delgado. En la cavidad oral, se ha podido identificar la presencia de ACE2, principalmente, en células epiteliale s de glándulas salivales y células epiteliales de la lengua. Además, se ha reportado la manifestación de algunos síntomas, como sequedad bucal y ambligeustia, los que podrían estar relacionadas con una infección de SARS-CoV-2 en estos órganos. Sin embargo, son necesarios mayores estudios que evidencien esta situación.


ABSTRACT: A novel coronavirus (SARS-CoV-2) has been recognized as a etiologic agent of a mysterious pneumonia originating in Wuhan, China. WHO has named the new disease as COVID-19 and, in addition, has declared it a pandemic. Taxonomically, SARS-CoV-2 belongs to the betacoronavirus genus along with SARS-CoV and MERS-CoV. SARS-CoV-2 uses angiotensin-converting enzyme 2 (ACE2) as the target receptor for entry into a host cell. The expression of ACE2 in cells of human tissues could indicate a potential risk of recognition by the virus and, therefore, be susceptible to infection. Through some laboratory and bioinformatics techniques, high presence of ACE2 has been seen in type II alveolar epithelial cells of the lung and enterocytes of the small intestine. In oral cavity, mainly presence of ACE2 has been identified in epithelial cells of salivary glands and epithelial cells of tongue. In addition, manifestation of some symptoms, such as dry mouth and amblygeustia, have been reported, which could be related to a SARS-CoV-2 infection in these organs. However, further studies are needed to prove this situation.


Subject(s)
Humans , Angiotensin-Converting Enzyme Inhibitors , Coronavirus Infections/epidemiology , Peptidyl-Dipeptidase A/chemistry , Betacoronavirus/chemistry , Tissue Culture Techniques/methods , Alveolar Epithelial Cells/cytology , Alveolar Epithelial Cells/virology , Mouth/virology
6.
Article in English | WPRIM | ID: wpr-827259

ABSTRACT

BACKGROUND@#We previously demonstrated that continuous exposure to nitrous acid gas (HONO) for 4 weeks, at a concentration of 3.6 parts per million (ppm), induced pulmonary emphysema-like alterations in guinea pigs. In addition, we found that HONO affected asthma symptoms, based on the measurement of respiratory function in rats exposed to 5.8 ppm HONO. This study aimed to investigate the dose-response effects of HONO exposure on the histopathological alterations in the respiratory tract of guinea pigs to determine the lowest observed adverse effect level (LOAEL) of HONO.@*METHODS@#We continuously exposed male Hartley guinea pigs (n = 5) to four different concentrations of HONO (0.0, 0.1, 0.4, and 1.7 ppm) for 4 weeks (24 h/day). We performed histopathological analysis by observing lung tissue samples. We examined samples from three guinea pigs in each group under a light microscope and measured the alveolar mean linear intercept (Lm) and the thickness of the bronchial smooth muscle layer. We further examined samples from two guinea pigs in each group under a scanning electron microscope (SEM) and a transmission electron microscope (TEM).@*RESULTS@#We observed the following dose-dependent changes: pulmonary emphysema-like alterations in the centriacinar regions of alveolar ducts, significant increase in Lm in the 1.7 ppm HONO-exposure group, tendency for hyperplasia and pseudostratification of bronchial epithelial cells, and extension of the bronchial epithelial cells and smooth muscle cells in the alveolar duct regions.@*CONCLUSIONS@#These histopathological findings suggest that the LOAEL of HONO is < 0.1 ppm.


Subject(s)
Alveolar Epithelial Cells , Animals , Bronchi , Dose-Response Relationship, Drug , Emphysema , Epithelial Cells , Guinea Pigs , Hyperplasia , Inhalation Exposure , Lung , Pathology , Male , Microscopy, Electron, Scanning , Microscopy, Electron, Transmission , Myocytes, Smooth Muscle , Nitrous Acid , Toxicity
7.
Protein & Cell ; (12): 707-722, 2020.
Article in English | WPRIM | ID: wpr-827023

ABSTRACT

The 2019 novel coronavirus disease (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has occurred in China and around the world. SARS-CoV-2-infected patients with severe pneumonia rapidly develop acute respiratory distress syndrome (ARDS) and die of multiple organ failure. Despite advances in supportive care approaches, ARDS is still associated with high mortality and morbidity. Mesenchymal stem cell (MSC)-based therapy may be an potential alternative strategy for treating ARDS by targeting the various pathophysiological events of ARDS. By releasing a variety of paracrine factors and extracellular vesicles, MSC can exert anti-inflammatory, anti-apoptotic, anti-microbial, and pro-angiogenic effects, promote bacterial and alveolar fluid clearance, disrupt the pulmonary endothelial and epithelial cell damage, eventually avoiding the lung and distal organ injuries to rescue patients with ARDS. An increasing number of experimental animal studies and early clinical studies verify the safety and efficacy of MSC therapy in ARDS. Since low cell engraftment and survival in lung limit MSC therapeutic potentials, several strategies have been developed to enhance their engraftment in the lung and their intrinsic, therapeutic properties. Here, we provide a comprehensive review of the mechanisms and optimization of MSC therapy in ARDS and highlighted the potentials and possible barriers of MSC therapy for COVID-19 patients with ARDS.


Subject(s)
Adoptive Transfer , Alveolar Epithelial Cells , Pathology , Animals , Apoptosis , Betacoronavirus , Body Fluids , Metabolism , CD4-Positive T-Lymphocytes , Allergy and Immunology , Clinical Trials as Topic , Coinfection , Therapeutics , Coronavirus Infections , Allergy and Immunology , Disease Models, Animal , Endothelial Cells , Pathology , Extracorporeal Membrane Oxygenation , Genetic Therapy , Methods , Genetic Vectors , Therapeutic Uses , Humans , Immunity, Innate , Inflammation Mediators , Metabolism , Lung , Pathology , Mesenchymal Stem Cell Transplantation , Methods , Mesenchymal Stem Cells , Physiology , Multiple Organ Failure , Pandemics , Pneumonia, Viral , Allergy and Immunology , Respiratory Distress Syndrome , Allergy and Immunology , Pathology , Therapeutics , Translational Medical Research
8.
Protein & Cell ; (12): 707-722, 2020.
Article in English | WPRIM | ID: wpr-828750

ABSTRACT

The 2019 novel coronavirus disease (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has occurred in China and around the world. SARS-CoV-2-infected patients with severe pneumonia rapidly develop acute respiratory distress syndrome (ARDS) and die of multiple organ failure. Despite advances in supportive care approaches, ARDS is still associated with high mortality and morbidity. Mesenchymal stem cell (MSC)-based therapy may be an potential alternative strategy for treating ARDS by targeting the various pathophysiological events of ARDS. By releasing a variety of paracrine factors and extracellular vesicles, MSC can exert anti-inflammatory, anti-apoptotic, anti-microbial, and pro-angiogenic effects, promote bacterial and alveolar fluid clearance, disrupt the pulmonary endothelial and epithelial cell damage, eventually avoiding the lung and distal organ injuries to rescue patients with ARDS. An increasing number of experimental animal studies and early clinical studies verify the safety and efficacy of MSC therapy in ARDS. Since low cell engraftment and survival in lung limit MSC therapeutic potentials, several strategies have been developed to enhance their engraftment in the lung and their intrinsic, therapeutic properties. Here, we provide a comprehensive review of the mechanisms and optimization of MSC therapy in ARDS and highlighted the potentials and possible barriers of MSC therapy for COVID-19 patients with ARDS.


Subject(s)
Adoptive Transfer , Alveolar Epithelial Cells , Pathology , Animals , Apoptosis , Betacoronavirus , Body Fluids , Metabolism , CD4-Positive T-Lymphocytes , Allergy and Immunology , Clinical Trials as Topic , Coinfection , Therapeutics , Coronavirus Infections , Allergy and Immunology , Disease Models, Animal , Endothelial Cells , Pathology , Extracorporeal Membrane Oxygenation , Genetic Therapy , Methods , Genetic Vectors , Therapeutic Uses , Humans , Immunity, Innate , Inflammation Mediators , Metabolism , Lung , Pathology , Mesenchymal Stem Cell Transplantation , Methods , Mesenchymal Stem Cells , Physiology , Multiple Organ Failure , Pandemics , Pneumonia, Viral , Allergy and Immunology , Respiratory Distress Syndrome , Allergy and Immunology , Pathology , Therapeutics , Translational Medical Research
9.
Protein & Cell ; (12): 707-722, 2020.
Article in English | WPRIM | ID: wpr-828586

ABSTRACT

The 2019 novel coronavirus disease (COVID-19), caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has occurred in China and around the world. SARS-CoV-2-infected patients with severe pneumonia rapidly develop acute respiratory distress syndrome (ARDS) and die of multiple organ failure. Despite advances in supportive care approaches, ARDS is still associated with high mortality and morbidity. Mesenchymal stem cell (MSC)-based therapy may be an potential alternative strategy for treating ARDS by targeting the various pathophysiological events of ARDS. By releasing a variety of paracrine factors and extracellular vesicles, MSC can exert anti-inflammatory, anti-apoptotic, anti-microbial, and pro-angiogenic effects, promote bacterial and alveolar fluid clearance, disrupt the pulmonary endothelial and epithelial cell damage, eventually avoiding the lung and distal organ injuries to rescue patients with ARDS. An increasing number of experimental animal studies and early clinical studies verify the safety and efficacy of MSC therapy in ARDS. Since low cell engraftment and survival in lung limit MSC therapeutic potentials, several strategies have been developed to enhance their engraftment in the lung and their intrinsic, therapeutic properties. Here, we provide a comprehensive review of the mechanisms and optimization of MSC therapy in ARDS and highlighted the potentials and possible barriers of MSC therapy for COVID-19 patients with ARDS.


Subject(s)
Adoptive Transfer , Alveolar Epithelial Cells , Pathology , Animals , Apoptosis , Betacoronavirus , Body Fluids , Metabolism , CD4-Positive T-Lymphocytes , Allergy and Immunology , Clinical Trials as Topic , Coinfection , Therapeutics , Coronavirus Infections , Allergy and Immunology , Disease Models, Animal , Endothelial Cells , Pathology , Extracorporeal Membrane Oxygenation , Genetic Therapy , Methods , Genetic Vectors , Therapeutic Uses , Humans , Immunity, Innate , Inflammation Mediators , Metabolism , Lung , Pathology , Mesenchymal Stem Cell Transplantation , Methods , Mesenchymal Stem Cells , Physiology , Multiple Organ Failure , Pandemics , Pneumonia, Viral , Allergy and Immunology , Respiratory Distress Syndrome , Allergy and Immunology , Pathology , Therapeutics , Translational Medical Research
10.
Rev. chil. enferm. respir ; 35(4): 261-263, dic. 2019. graf
Article in Spanish | LILACS | ID: biblio-1092702

ABSTRACT

La fibrosis pulmonar idiopática (FPI) es una forma específica de neumonía intersticial idiopática, de tipo fibrosante crónica y progresiva, con patrón radiológico y/o histológico de neumonía intersticial usual (NIU). Su patogenia es compleja, el modelo más aceptado actualmente es basado en las células epiteliales alveolares, aberrantemente activadas que conducen a la proliferación de fibroblastos y su diferenciación a miofibroblastos que depositan matriz extracelular y destruyen irreversiblemente la arquitectura pulmonar. No existe un claro factor inicial que explique la activación y posterior mantención del mecanismo de la fibrosis. El factor de crecimiento transformante beta (TGF-β) liberado por las células epiteliales alveolares se ha implicado como unos de los principales conductores de la inducción y proliferación de fibroblastos alterados que persiste mucho tiempo después de la estimulación inicial, lo que explicaría en gran parte el comportamiento clínico progresivo y crónico.


Idiopathic pulmonary fibrosis (IPF) is a specific form of idiopathic interstitial pneumonia, of chronic and progressive fibrosing type, with radiological and / or histological pattern of usual interstitial pneumonia (UIP). Its pathogenesis is complex, the most accepted model currently is based on the fact that the alveolar epithelial cells, aberrantly activated, lead to the proliferation of fibroblasts and their differentiation to myofibroblasts that deposit extracellular matrix and irreversibly destroy the pulmonary architecture. There is no clear initial trigger that explains the activation and subsequent maintenance of the fibrosis mechanism. The transforming growth factor beta (TGF-β), released by the alveolar epithelial cells, has been implicated as one of the main drivers of the induction and proliferation of altered fibroblasts that persists long after the initial stimulation, which would largely explain progressive and chronic clinical behavior.


Subject(s)
Humans , Idiopathic Pulmonary Fibrosis/etiology , Idiopathic Pulmonary Fibrosis/physiopathology , Idiopathic Pulmonary Fibrosis/epidemiology , Risk Factors , Transforming Growth Factor beta , Extracellular Matrix , Alveolar Epithelial Cells
11.
Acta Physiologica Sinica ; (6): 575-580, 2019.
Article in Chinese | WPRIM | ID: wpr-777154

ABSTRACT

The aim of the present study was to investigate the effect of salidroside (Sal) on inflammatory activation induced by lipopolysaccharide (LPS) in the co-culture of rat alveolar macrophages (AM) NR 8383 and type II alveolar epithelial cells (AEC II) RLE-6TN. CCK-8 colorimetric method was used to detect cell proliferation percentage. The enzyme-linked immunosorbent assay (ELISA) was used to determine the content of tumor necrosis factor alpha (TNF-α), macrophage inflammatory protein-2 (MIP-2) and interleukin-10 (IL-10) in the supernatant. Western blot was used to examine the expression levels of phosphorylated AKT (p-AKT) and total AKT protein. The results showed that pretreatment of RLE-6TN cells or co-culture of RLE-6TN and NR 8383 cells with 32 and 128 µg/mL Sal for 1 h, followed by continuous culture for 24 h, significantly increased the cell proliferation (P < 0.05). Compared with control group, 32 and 128 µg/mL Sal pretreatment significantly increased the ratio of p-AKT/AKT in RLE-6TN cells (P < 0.05). Pretreatment of 32 µg/mL Sal not only inhibited the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05), but also enhanced the inhibitory effect of RLE-6TN and NR 8383 cells co-culture on the secretion of TNF-α and MIP-2 by NR 8383 cells induced by LPS (P < 0.05). In addition, 32 µg/mL Sal pretreatment promoted LPS-induced IL-10 secretion by NR 8383 cells (P < 0.05), and enhanced the promoting effect of co-culture of RLE-6TN and NR 8383 cells on the IL-10 secretion by LPS-induced NR 8383 cells (P < 0.05). In conclusion, Sal may directly inhibit LPS-induced inflammatory activation of AM (NR 8383), promote the proliferation of AEC II (RLE-6TN) through PI3K/AKT signaling pathway, and enhance the regulatory effect of AEC II on LPS-induced inflammatory activation of AM.


Subject(s)
Alveolar Epithelial Cells , Metabolism , Animals , Cell Line , Chemokine CXCL2 , Metabolism , Coculture Techniques , Glucosides , Pharmacology , Interleukin-10 , Metabolism , Lipopolysaccharides , Macrophages, Alveolar , Metabolism , Phenols , Pharmacology , Phosphatidylinositol 3-Kinases , Metabolism , Proto-Oncogene Proteins c-akt , Metabolism , Rats , Signal Transduction , Tumor Necrosis Factor-alpha , Metabolism
12.
Braz. j. med. biol. res ; 51(2): e6950, 2018. tab, graf
Article in English | LILACS | ID: biblio-889028

ABSTRACT

Alveolar epithelia play an essential role in maintaining the integrity and homeostasis of lungs, in which alveolar epithelial type II cells (AECII) are a cell type with stem cell potential for epithelial injury repair and regeneration. However, mechanisms behind the physiological and pathological roles of alveolar epithelia in human lungs remain largely unknown, partially owing to the difficulty of isolation and culture of primary human AECII cells. In the present study, we aimed to characterize alveolar epithelia generated from A549 lung adenocarcinoma cells that were cultured in an air-liquid interface (ALI) state. Morphological analysis demonstrated that A549 cells could reconstitute epithelial layers in ALI cultures as evaluated by histochemistry staining and electronic microscopy. Immunofluorescent staining further revealed an expression of alveolar epithelial type I cell (AECI) markers aquaporin-5 protein (AQP-5), and AECII cell marker surfactant protein C (SPC) in subpopulations of ALI cultured cells. Importantly, molecular analysis further revealed the expression of AQP-5, SPC, thyroid transcription factor-1, zonula occludens-1 and Mucin 5B in A549 ALI cultures as determined by both immunoblotting and quantitative RT-PCR assay. These results suggest that the ALI culture of A549 cells can partially mimic the property of alveolar epithelia, which may be a feasible and alternative model for investigating roles and mechanisms of alveolar epithelia in vitro.


Subject(s)
Humans , Culture Media, Conditioned , Cell Culture Techniques/methods , Alveolar Epithelial Cells/physiology , A549 Cells/physiology , Reference Values , Time Factors , Microscopy, Electron, Scanning , Immunoblotting , Cell Count , Reproducibility of Results , Analysis of Variance , Pulmonary Surfactant-Associated Protein C/analysis , Aquaporin 5/analysis , Mucin-5B/analysis , Real-Time Polymerase Chain Reaction , Zonula Occludens-1 Protein/analysis , Thyroid Nuclear Factor 1/analysis
13.
Rev. bras. anestesiol ; 67(6): 600-606, Nov.-Dec. 2017. graf
Article in English | LILACS | ID: biblio-897789

ABSTRACT

Abstract Background and objectives Dexmedetomidine (DEX) has demonstrated the preconditioning effect and shown protective effects against organize injury. In this study, using A549 (human alveolar epithelial cell) cell lines, we investigated whether DEX preconditioning protected against acute lung injury (ALI) in vitro. Methods A549 were randomly divided into four groups (n = 5): control group, DEX group, lipopolysaccharides (LPS) group, and D-LPS (DEX + LPS) group. Phosphate buffer saline (PBS) or DEX were administered. After 2 h preconditioning, the medium was refreshed and the cells were challenged with LPS for 24 h on the LPS and D-LPS group. Then the malondialdehyde (MDA), superoxide dismutase (SOD), Bcl-2, Bax, caspase-3 and the cytochrome c in the A549 were tested. The apoptosis was also evaluated in the cells. Results Compare with LPS group, DEX preconditioning reduced the apoptosis (26.43% ± 1.05% vs. 33.58% ± 1.16%, p < 0.05) in the A549, which is correlated with decreased MDA (12.84 ± 1.05 vs. 19.16 ± 1.89 nmoL.mg-1 protein, p < 0.05) and increased SOD activity (30.28 ± 2.38 vs. 20.86 ± 2.19 U.mg-1 protein, p < 0.05). DEX preconditioning also increased the Bcl-2 level (0.53 ± 0.03 vs. 0.32 ± 0.04, p < 0.05) and decreased the level of Bax (0.49 ± 0.04 vs. 0.65 ± 0.04, p < 0.05), caspase-3 (0.54 ± 0.04 vs. 0.76 ± 0.04, p < 0.05) and cytochrome c. Conclusion DEX preconditioning has a protective effect against ALI in vitro. The potential mechanisms involved are the inhibition of cell death and improvement of antioxidation.


Resumo Justificativa e objetivos Dexmedetomidina (DEX) demonstrou ter efeito pré-condicionante e também efeitos protetores contra lesão organizada. Neste estudo, com células A549 (células epiteliais alveolares humanas), investigamos se o pré-condicionamento com DEX proporcionaria proteção contra lesão pulmonar aguda (LPA) in vitro. Métodos Células A549 foram aleatoriamente distribuídas em quatro grupos (n = 5): controle, DEX, lipopolissacarídeos (LPS) e D-LPS (DEX + LPS). Administramos solução de PBS (tampão fosfato-alcalino) ou DEX. Após 2 h de pré-condicionamento, o meio foi renovado e as células desafiadas com LPS por 24 h nos grupos LPS e D-LPS. Em seguida, malondialdeído (MDA), superóxido dismutase (SOD), Bcl-2, Bax, caspase-3 e em A549 foram testados. Apoptose também foi avaliada nas células. Resultados Em comparação com o grupo LPS, o pré-condicionamento com DEX reduziu a apoptose (26,43% ± 1,05% vs. 33,58% ± 1,16%, p < 0,05) em células A549, o que está correlacionado com a diminuição de MDA (12,84 ± 1,05 vs. 19,16 ± 1,89 nmol.mg-1 de proteína, p < 0,05) e aumento da atividade de SOD (30,28 ± 2,38 vs. 20,86 ± 2,19 U.mg-1 de proteína, p < 0,05). O pré-condicionamento com DEX também aumentou o nível de Bcl-2 (0,53 ± 0,03 vs. 0,32 ± 0,04, p < 0,05) e diminuiu o nível de Bax (0,49 ± 0,04 vs. 0,65 ± 0,04, p < 0,05), caspase-3 (0,54 ± 0,04 vs. 0,76 ± 0,04, p < 0,05) e citocromo c. Conclusão O pré-condicionamento com DEX tem efeito protetor contra LPA in vitro. Os potenciais mecanismos envolvidos são inibição da morte celular e melhoria da antioxidação.


Subject(s)
Humans , Lipopolysaccharides/adverse effects , Dexmedetomidine/pharmacology , Alveolar Epithelial Cells/drug effects , Hypnotics and Sedatives/pharmacology , Random Allocation , Cells, Cultured , Lipopolysaccharides/antagonists & inhibitors
14.
Arq. bras. med. vet. zootec. (Online) ; 69(3): 593-596, jun. 2017. ilus
Article in Portuguese | LILACS, VETINDEX | ID: biblio-846892

ABSTRACT

As doenças pulmonares intersticiais constituem um grupo de doenças difusas do parênquima pulmonar, no qual a fibrose pulmonar intersticial está incluída. Histologicamente, esta se caracteriza por hiperplasia de pneumócitos tipo II, hiperplasia ou hipertrofia de músculo liso e fibrose. Embora a patogenia da fibrose pulmonar intersticial não esteja bem elucidada, devido às semelhanças microscópicas encontradas nos pneumócitos tipo II em felinos e na forma familiar da doença em humanos, acredita-se que haja caráter genético para o seu desenvolvimento. Os sinais clínicos frequentemente relatados incluem desconforto respiratório, cianose, letargia e perda de peso. Devido ao caráter progressivo e à ausência de tratamento específico, a doença apresenta prognóstico desfavorável. Foi atendida uma gata de 12 anos de idade, com histórico de dispneia há 20 dias. Ao exame clínico, o animal apresentou dispneia expiratória restritiva, crepitação à ausculta torácica e foi visualizado padrão intersticial ao exame radiográfico do tórax. A paciente foi submetida à punção com agulha fina de tecido pulmonar e veio a óbito algumas horas após o procedimento, apresentando insuficiência respiratória aguda. No exame histológico do tecido pulmonar, foi verificada a ocorrência de fibrose pulmonar idiopática. O objetivo do presente trabalho é relatar um caso de dispneia expiratória restritiva em um felino doméstico devido à fibrose pulmonar idiopática, já que, segundo o conhecimento dos autores, não há nenhum relato da ocorrência da doença no país.(AU)


Interstitial lung diseases are a group of diffuse parenchymal lung diseases in which interstitial lung fibrosis is included. Histologically, it is characterized by type II pneumocyte hyperplasia, hypertrophy or hyperplasia of smooth tissue and fibrosis. Although the pathogenesis of interstitial lung fibrosis has not been elucidated, due to the microscopic similarities found in type II pneumocytes in cats and familial form of the disease in humans, it is believed that there is a genetic trait for development. The frequently reported clinical signs include respiratory distress, cyanosis, lethargy, and weight loss. Due to the progressive nature and the absence of specific treatment, the disease has a poor prognosis. A 12-year-old cat with dyspnea for 20 days was assisted. The animal underwent fine needle aspiration of lung tissue and died few hours after the procedure, with acute respiratory failure. Upon histological examination of lung tissue, the occurrence of idiopathic pulmonary fibrosis was found. The aim of this study is to report a case of restrictive expiratory dyspnea in a domestic feline due to idiopathic pulmonary fibrosis, because, according to our knowledge, there is no report on the occurrence of the disease in our country.(AU)


Subject(s)
Animals , Cats , Dyspnea/veterinary , Idiopathic Pulmonary Fibrosis/veterinary , Alveolar Epithelial Cells , Lung Diseases, Interstitial/veterinary
16.
Article in Chinese | WPRIM | ID: wpr-236385

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the effects of ischemic postconditioning (IPostC) on pneumocyte apoptosis after lung ischemia/reperfusion injury in rats.</p><p><b>METHODS</b>Adult male SD rats were randomly divided into 3 groups based upon the intervention (n = 8): control group (C), lung ischemic reperfusion group (LIR), LIR+ IPostC group (IPostC). At the end of the experiment, blood specimens drawn from the arteria carotis were tested for the content of malondialdehyde (MDA), the activity of superoxide dismutase (SOD) and myeloperoxidase (MPO); the pneumocyte apoptosis index (AI) was achieved by tennrminal deoxynucleotidyl transferase mediated dUTP nick end abeling (TUNEL); the expression of Bcl-2, Bax protein in lung tissue was accessed by quantitative immunohistochemistry (MHC) and Bcl-2, Bax mRNA by RT-PCR.</p><p><b>RESULTS</b>IPostC could significantly attenuate the MDA level, MPO activity and improve SOD activity in blood serum which was comparable to I/R and significantly reduced the number of TUNEL-positive cells compared with I/R group, expressed as Al (% total nuclei) from (39.0 +/- 3.46) to (8.0 +/- 0.88) (P < 0.01). The protein and mRNA expression of Bcl-2 and Bax showed that IPO significantly attenuated the ischemia/reperfusion-upregulated expression of Bax protein but improved the expression of Bcl-2 that improved the Bcl-2/Bax ratio (P < 0.01) .</p><p><b>CONCLUSION</b>IPostC may attenuate pneumocyte apoptosis in LIRI by up-regulating expression of Bcl-2/Bax ratio and by inhibiting oxidant generation and neutrophils filtration.</p>


Subject(s)
Alveolar Epithelial Cells , Cell Biology , Animals , Apoptosis , Ischemic Postconditioning , Lung , Metabolism , Pathology , Lung Injury , Male , Malondialdehyde , Metabolism , Peroxidase , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Rats , Rats, Sprague-Dawley , Reperfusion Injury , Superoxide Dismutase , Metabolism , bcl-2-Associated X Protein , Metabolism
17.
Article in Chinese | WPRIM | ID: wpr-236333

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the role of p38 MAPK on ischemic postconditioning (IPO) attenuating pneumocyte apoptosis after lung ischemia/reperfusion injury (LIRI).</p><p><b>METHODS</b>Forty adult male SD rats were randomly divided into 5 groups based upon the intervention (n = 8): control group (C), LIR group (I/R), LIR + IPO group (IPO), IPO + solution control group (D), IPO + SB203580 group (SB). Left lung tissue was isolated after the 2 hours of reperfusion, the ratio of wet lung weight to dry lung weight (W/D), and total lung water content (TLW) were measured. The histological structure of the left lung was observed under light and electron transmission microscopes, and scored by alveolar damage index of quantitative assessment (IQA). Apoptosis index (AI) of lung tissue was determined by terminal deoxynuleotidyl transferase mediated dUTP nick end and labeling (TUNEL) method. The mRNA expression and protein levels of and Bax were measured by RT-PCR and quantitative immunohistochemistry (IHC).</p><p><b>RESULTS</b>Compared with C group, W/D, TLW, IQA, AI and the expression of Bax of I/R were significantly increased, the expression of Bcl-2 and Bcl-2/Bax were significantly decreased (P < 0.05, P < 0.01), and was obviously morphological abnormality in lung tissue. Compared with I/R group, all the indexes of IPO except for the expression of Bcl-2 and Bcl-2/ Bax were obviously reduced, the expression of Bcl-2 and Bcl-2/Bax were increased (P < 0.05, P < 0.01). All the indexes between D and IPO were little or not significant( P > 0.05). The expression of Bcl-2 and Bcl-2/Bax of SB were significantly increased and other indexes were reduced than those of IPO (P < 0.05, P < 0.01).</p><p><b>CONCLUSION</b>IPO may attenuate pneumocyte apoptosis in LIRI by inactivation of p38 MAPK, up-regulating expression of Bcl-2/Bax ratio.</p>


Subject(s)
Alveolar Epithelial Cells , Cell Biology , Animals , Apoptosis , Disease Models, Animal , Ischemic Postconditioning , Lung , Pathology , Male , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Rats , Rats, Sprague-Dawley , Reperfusion Injury , Pathology , bcl-2-Associated X Protein , Metabolism , p38 Mitogen-Activated Protein Kinases , Metabolism
18.
Korean Journal of Medicine ; : 623-626, 2014.
Article in Korean | WPRIM | ID: wpr-151952

ABSTRACT

Alveolar adenoma is a very rare benign intraparenchymal lung tumor originating from type II pneumocytes. It can be mistaken for other benign tumors or lung cancer in radiological images. It is especially difficult to distinguish alveolar adenoma from sclerosing hemangioma. A small aspiration biopsy specimen, such as with percutaneous needle aspiration, is insufficient for a pathological diagnosis. Surgical resection is the only method by which a pathological diagnosis can be made and the disease treated. An alveolar adenoma presenting as multiple nodules is very rare and has to our knowledge not been reported in Korea previously. Here, we report a case of alveolar adenoma in multiple nodules in a 57-year-old female and review the literature.


Subject(s)
Adenoma , Biopsy, Needle , Diagnosis , Female , Histiocytoma, Benign Fibrous , Humans , Korea , Lung Neoplasms , Lung , Middle Aged , Needles , Alveolar Epithelial Cells
19.
Article in English | WPRIM | ID: wpr-31121

ABSTRACT

BACKGROUND AND OBJECTIVES: The fibrosing form of lung injury (occupational, environmental, infective or drug induced) is associated with significant morbidity and mortality. Amiodarone (AM), often prescribed for control of arrhythmias is considered a potential cause. No effective treatment was confirmed, except lung transplantation. Intravenous (IV) stem cell therapy may produce pulmonary emboli or infarctions. Despite being commonly used in clinical practice, the intraperitoneal (IP.) route has been rarely used for cell delivery. The present study aimed at investigating and comparing the possible effect of IP stem cell therapy (SCT) on pulmonary toxicity versus the intravenous route in a rat model of amiodarone induced lung damage. METHODS AND RESULTS: 36 adult male albino rats were divided into 4 groups. Rats of AM group were given 30 mg/kg daily orally for 4 weeks. Rats of IV SCT group were injected with stem cells in the tail vein. Rats of IP SCT group received IP cell therapy. Histological, histochemical, immunohistochemical and morphometric studies were performed. Obstructed bronchioles, overdistended alveoli, reduced type I pneumocytes, increased thickness of alveolar septa and vessels wall besides increased area% of collagen fibers regressed in response to IV and IP SCT. The improvement was more obvious in IV group. The area% of Prussion blue +ve and CD105 +ve cells was significantly higher in IV group. CONCLUSIONS: Cord blood MSC therapy proved definite amelioration of lung injury ending in fibrosis. The effect of IP SCT was slightly inferior to that of IV SCT, which may be overwhelmed by repeated IP injection.


Subject(s)
Adult , Amiodarone , Animals , Arrhythmias, Cardiac , Bronchioles , Cell- and Tissue-Based Therapy , Collagen , Fetal Blood , Fibrosis , Humans , Infarction , Lung , Lung Injury , Lung Transplantation , Male , Mesenchymal Stem Cells , Models, Animal , Mortality , Alveolar Epithelial Cells , Rats , Stem Cells , Veins
20.
Article in English | WPRIM | ID: wpr-31117

ABSTRACT

AIM OF WORK: To demonstrate the bleomycin induced histological changes in the lung and the possible protective and/or therapeutic effect of stem cell therapy. MATERIALS AND METHODS: Study was carried out on 36 adult male albino rats, classified into 4 groups: group I (control), group II (bleomycin treated group), group III (early stem cell treated group: immediately after bleomycin), group IV (late stem cell treated group: 7 days after bleomycin). Sections were taken at the 14th day of experiment. stained with Hematoxylin and Eosin, Masson's trichrome, immunohistochemichal stains for alpha-SMA & PCNA. Sections were examined by light & immunofluroscent microscopy. Area percent of collagen fibers, area percent & optical density of alpha-SMA immunopositive cells were measured as well as the number of H&E and PCNA stained pneumocytes type II was counted. RESULTS: Group II showed, thickening of septa, extravasation of blood, dividing pneumocytes type II cells with acinar formation, cellular infiltration, fibroblast cells, almost complete loss of normal lung architecture in certain fields, consolidation and replacement of the lung tissue with fibrous tissue in other fields. Restoring of lung tissue with significant decrease in mean area % of collagen fibers, alpha-SMA immunopositive cells were detected in group III. CONCLUSIONS: Early treatment with bone marrow derived mesenchymal stem cells (BMSCs) immediately after bleomycin administration showed a significant reduction in fibrotic changes, however the late treatment with BMSCs (7 days) after bleomycin administration showed non significant results.


Subject(s)
Adult , Animals , Bleomycin , Bone Marrow , Collagen , Coloring Agents , Eosine Yellowish-(YS) , Fibroblasts , Hematoxylin , Humans , Lung , Male , Mesenchymal Stem Cells , Microscopy , Alveolar Epithelial Cells , Proliferating Cell Nuclear Antigen , Pulmonary Fibrosis , Rats , Stem Cells
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