Monocyte inflammation augments acrolein-induced Muc5ac expression in mouse lung

1999 ◽  
Vol 277 (3) ◽  
pp. L489-L497 ◽  
Author(s):  
Michael T. Borchers ◽  
Scott Wesselkamper ◽  
Susan E. Wert ◽  
Steven D. Shapiro ◽  
George D. Leikauf
Keyword(s):  
Gene Expression ◽  
Neutrophil Elastase ◽  
Airway Hyperreactivity ◽  
Mouse Lung ◽  
Mrna Levels ◽  
Elastase Activity ◽  
Mucus Hypersecretion ◽  
Mucin Gene ◽  
Cell Counts ◽  
Macrophage Accumulation

Acrolein, an unsaturated aldehyde found in smog and tobacco smoke, can induce airway hyperreactivity, inflammation, and mucus hypersecretion. To determine whether changes in steady-state mucin gene expression ( Muc2 and Muc5ac) are associated with inflammatory cell accumulation and neutrophil elastase activity, FVB/N mice were exposed to acrolein (3.0 parts/million; 6 h/day, 5 days/wk for 3 wk). The levels of Muc2 and Muc5ac mRNA were determined by RT-PCR, and the presence of Muc5ac protein was detected by immunohistochemistry. Total and differential cell counts were determined from bronchoalveolar lavage (BAL) fluid, and neutrophil elastase activity was measured in the BAL fluid supernatant. Lung Muc5ac mRNA was increased on days 12and 19, and Muc5ac protein was detected in mucous granules and on the surface of the epithelium on day 19. Lung Muc2 mRNA was not detected at measurable levels in either control or exposed mice. Acrolein exposure caused a significant and persistent increase in macrophages and a rapid but transient increase in neutrophils in BAL fluid. Recoverable neutrophil elastase activity was not significantly altered at any time after acrolein exposure. To further examine the role of macrophage accumulation in mucin gene expression, additional strains of mice (including a strain genetically deficient in macrophage metalloelastase) were exposed to acrolein for 3 wk, and Muc5ac mRNA levels and macrophage accumulation were measured. The magnitude of macrophage accumulation coincided with increased Muc5ac mRNA levels, indicating that excessive macrophage accumulation augments acrolein-induced Muc5ac synthesis and secretion after repeated exposure. These findings support a role for chronic monocytic inflammation in the pathogenesis of mucus hypersecretion observed in chronic bronchitis.

scholarly journals Neutrophil elastase increasesMUC5ACmRNA and protein expression in respiratory epithelial cells

1999 ◽  
Vol 276 (5) ◽  
pp. L835-L843 ◽  
Author(s):  
Judith A. Voynow ◽  
Lisa Rosenthal Young ◽  
Yiqiong Wang ◽  
Teresa Horger ◽  
Mary C. Rose ◽  
...  
Keyword(s):  
Gene Expression ◽  
Gene Regulation ◽  
Epithelial Cells ◽  
Serine Protease ◽  
Neutrophil Elastase ◽  
A549 Cells ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Mucin Production ◽  
Mucin Gene

Chronic neutrophil-predominant inflammation and hypersecretion of mucus are common pathophysiological features of cystic fibrosis, chronic bronchitis, and viral- or pollution-triggered asthma. Neutrophils release elastase, a serine protease, that causes increased mucin production and secretion. The molecular mechanisms of elastase-induced mucin production are unknown. We hypothesized that as part of this mechanism, elastase upregulates expression of a major respiratory mucin gene, MUC5AC. A549, a human lung carcinoma cell line that expresses MUC5AC mRNA and protein, and normal human bronchial epithelial cells in an air-liquid interface culture were stimulated with neutrophil elastase. Neutrophil elastase increased MUC5AC mRNA levels in a time-dependent manner in both cell culture systems. Neutrophil elastase treatment also increased MUC5AC protein levels in A549 cells. The mechanism of MUC5AC gene regulation by elastase was determined in A549 cells. The induction of MUC5AC gene expression required serine protease activity; other classes of proteases had no effect on MUC5AC gene expression. Neutrophil elastase increased MUC5AC mRNA levels by enhancing mRNA stability. This is the first report of mucin gene regulation by this mechanism.

Acrolein-induced MUC5ac expression in rat airways

1998 ◽  
Vol 274 (4) ◽  
pp. L573-L581 ◽  
Author(s):  
Michael T. Borchers ◽  
Susan E. Wert ◽  
George D. Leikauf
Keyword(s):  
Gene Expression ◽  
Immunohistochemical Analysis ◽  
Airway Hyperreactivity ◽  
Chronic Obstructive ◽  
Mrna Levels ◽  
Temporal Expression ◽  
Histochemical Analysis ◽  
Obstructive Pulmonary Disease ◽  
Mucus Hypersecretion ◽  
Mucin Glycoproteins

Acrolein, a low-molecular-weight aldehyde found in photochemical smog and tobacco smoke, can induce mucus hypersecretion, inflammation, and airway hyperreactivity. To determine whether changes in steady-state mucin gene expression (MUC2 and MUC5ac) are associated with histological signs of mucus hypersecretion, rats were exposed to acrolein (3.0 parts/million, 6 h/day, 5 days/wk, 2 wk), and the trachea with the main stem bronchi was separated from the intrapulmonary airways (lung). The temporal expression of MUC2 and MUC5ac mRNA was determined by RT-PCR, and acidic mucin glycoproteins were detected by Alcian blue histochemical analysis. MUC5ac protein content in the airways was determined by immunohistochemical analysis. Tracheal MUC5ac mRNA increased within 2 days and was accompanied by an increase in MUC5ac immunostaining on the surface of the airways and in submucosal gland epithelium. By comparison, increases in lung MUC5ac mRNA and mucin glycoproteins were delayed and were elevated after exposures on days 5 and 9, respectively. Increased MUC5ac immunostaining was detected within the lumen and airway epithelium of the lung on day 12. In contrast, MUC2 mRNA levels were not significantly changed in the trachea or lung. These findings indicate that acrolein-induced mucus hypersecretion is due, in part, to increases in MUC5ac rather than to MUC2 gene expression. These findings suggest that aldehyde-induced increases in MUC5ac may play a role in chronic mucus hypersecretion, a pathognomonic feature of chronic obstructive pulmonary disease.

scholarly journals Transcriptional and posttranscriptional modulation of human neutrophil elastase gene expression

Blood ◽  
1992 ◽  
Vol 79 (10) ◽  
pp. 2733-2740 ◽  
Author(s):  
K Yoshimura ◽  
RG Crystal
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Gene Transcription ◽  
Neutrophil Elastase ◽  
Human Neutrophil ◽  
Leukemia Cell Line ◽  
Human Neutrophil Elastase ◽  
Mrna Levels ◽  
Mrna Transcript ◽  
Lineage Differentiation

Abstract Human neutrophil elastase (NE), a 29-Kd potent serine protease stored in azurophilic granules of mature neutrophils, is coded for by the NE gene, a single copy gene with 5 exons spanning a 6-kb segment of chromosome 11 at q14. With the knowledge that the NE gene expression is limited to early myeloid cell differentiation, mechanisms modulating expression of the NE gene were evaluated in the HL-60 promyelocytic leukemia cell line, a model of early bone marrow precursor cells. Consistent with the presence of NE messenger RNA (mRNA) transcripts in undifferentiated HL-60 cells, nuclear transcription run-on analyses showed that HL-60 cells actively transcribed the NE gene. However, the transcription rate of the NE gene was relatively low, only 40% of the myeloperoxidase gene, a gene expressed in parallel with NE. When induced toward the mononuclear phagocytic lineage with phorbol 12- myristate 13-acetate (PMA), HL-60 cells exhibited marked suppression of NE gene transcription, declining to 17% of the resting rate within 2 days. Induction toward mononuclear phagocytic lineage differentiation caused no change in NE mRNA transcript half-life (T1/2), but mRNA levels decreased markedly over time, with levels undetectable 1.5 days after PMA stimulation. In contrast, when induced toward the myelocytic lineage with dimethyl sulfoxide, the rate of NE gene transcription increased 1.9-fold within 5 days. Interestingly, the mRNA transcript levels increased 2.5-fold by 5 days despite the fact that induction toward myelocytic lineage differentiation was accompanied by a marked reduction of NE mRNA transcript T1/2. Together, these observations suggest that the NE gene expression during bone marrow differentiation is modulated mainly at the transcriptional level, with some posttranscriptional modulation contributing, particularly during myelocytic lineage differentiation.

scholarly journals Amelioration of Cigarette Smoke-Induced Mucus Hypersecretion and Viscosity by Dendrobium officinale Polysaccharides In Vitro and In Vivo

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Rui Chen ◽  
Yingmin Liang ◽  
Mary Sau Man Ip ◽  
Kalin Yanbo Zhang ◽  
Judith Choi Wo Mak
Keyword(s):  
Gene Expression ◽  
Cigarette Smoke ◽  
Secretory Granules ◽  
Dendrobium Officinale ◽  
In Vivo Models ◽  
Mucus Hypersecretion ◽  
Mucin Gene ◽  
Starting Point

Chronic obstructive pulmonary disease (COPD), characterized by oxidative stress and inflammation, is one of the leading causes of death worldwide, in which cigarette smoke (CS) is the major risk factor. Dendrobium officinale polysaccharides (DOPs) are the main active ingredients extracted from Dendrobium officinale, which have been reported to have antioxidant and anti-inflammatory activity as well as inhibition of mucin gene expression. This study is aimed at investigating the effect of DOPs on CS-induced mucus hypersecretion and viscosity in vitro and in vivo. For in vitro study, primary normal human bronchial epithelial cells (HBECs) differentiated at the air-liquid interface (ALI) culture for 28 days were stimulated with cigarette smoke medium (CSM) in the absence or presence of various concentrations of DOPs or N-acetylcysteine (NAC) for 24 hours. For in vivo study, male Sprague-Dawley rats were randomized to sham air (SA) as control group or CS group for 56 days. At day 29, rats were subdivided and given water as control, DOPs, or NAC as positive control as a mucolytic drug via oral gavage for the remaining duration. Samples collected from apical washing, cell lysates, bronchoalveolar lavage (BAL), and lung tissues were evaluated for mucin gene expression, mucus secretion, and viscosity. DOPs ameliorated the CS-induced mucus hypersecretion and viscosity as shown by the downregulation of MUC5AC mRNA, MUC5AC secretary protein, and mucus viscosity via inhibition of mucus secretory granules in both in vitro and in vivo models. DOPs produced its effective effects on the CS-induced mucus hypersecretion and viscosity via the inhibition of the mucus secretory granules. These findings could be a starting point for considering the potential role of DOPs in the management of the smoking-mediated COPD. However, further research is needed.

Induction of mucin gene expression in human colonic cell lines by PMA is dependent on PKC-ε

1999 ◽  
Vol 277 (5) ◽  
pp. G1041-G1047 ◽  
Author(s):  
D.-H. Hong ◽  
G. Petrovics ◽  
W. B. Anderson ◽  
J. Forstner ◽  
G. Forstner
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Dominant Negative ◽  
Mrna Levels ◽  
Wild Type ◽  
Mucin Gene ◽  
Time Period ◽  
Mucin Expression ◽  
Mucin Genes

Treatment of HT-29 cells with phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C (PKC), induces MUC2 expression. To investigate the role of PKC in regulating mucin genes in intestinal cells, we examined the regulation of MUC1, MUC2, MUC5AC, MUC5B, and MUC6 expression in two human mucin-producing colonic cell lines, T84 and HT29/A1. T84 and HT29/A1 cells (at 80–90% confluency) were exposed to 100 nM PMA for 0, 3, and 6 h. Twofold or greater increases in mRNA levels for MUC2 and MUC5AC were observed in both cell lines during this time period, whereas the levels of MUC1, MUC5B, and MUC6 mRNAs were only marginally affected. These results indicated that PKC differentially regulates mucin gene expression and that it may be responsible for altered mucin expression. Our previous results suggested that the Ca2+-independent PKC-ε isoform appeared to mediate PMA-regulated mucin exocytosis in these cell lines. To determine if PKC-ε was also involved in MUC2/MUC5AC gene induction, HT29/A1 cells were stably transfected with either a wild-type PKC-ε or a dominant-negative ATP-binding mutant of PKC-ε (PKC-ε K437R). Overexpression of the dominant-negative PKC-ε K437R blocked induction of both mucin genes, whereas PMA-induced mucin gene expression was not prevented by overexpression of wild-type PKC-ε. PMA-dependent MUC2 mucin secretion was also blocked in cells overexpressing the dominant-negative PKC-ε K437R. On the basis of these observations, PKC-ε appears to mediate the expression of two major gastrointestinal mucins in response to PMA as well as PMA-regulated mucin exocytosis.

scholarly journals Changes in airway inflammation during pulmonary exacerbations in patients with cystic fibrosis and primary ciliary dyskinesia

2015 ◽  
Vol 47 (3) ◽  
pp. 829-836 ◽  
Author(s):  
Felix Ratjen ◽  
Valerie Waters ◽  
Michelle Klingel ◽  
Nancy McDonald ◽  
Sharon Dell ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Inflammatory Response ◽  
Neutrophil Elastase ◽  
Primary Ciliary Dyskinesia ◽  
Bacterial Pathogens ◽  
Pulmonary Exacerbation ◽  
Elastase Activity ◽  
Cell Counts ◽  
Pulmonary Exacerbations ◽  
Ciliary Dyskinesia

Lung disease in patients with both primary ciliary dyskinesia (PCD) or cystic fibrosis (CF) is associated with impaired mucociliary clearance; however, clinical outcomes are typically worse in CF patients. We assessed whether CF and PCD patients differ in inflammatory response in the airways during pulmonary exacerbation.We first studied clinically stable PCD patients with a spectrum of bacterial pathogens to assess inflammatory response to different pathogens. Subsequently, PCD and CF patients with similar bacterial pathogens were studied at the time of a pulmonary exacerbation and after 21 days of antibiotics treatment. Qualitative and quantitative microbiology, cell counts, interleukin-8 concentrations, and neutrophil elastase activity were assessed in sputum samples obtained before and after treatment.In stable PCD patients, no significant differences were found in sputum inflammatory markers between individuals colonised with different bacterial pathogens. Pulmonary exacerbation severity assessed by a pulmonary exacerbation score and lung function decline from their previous baseline did not differ between CF and PCD patients. Bacterial density for Staphylococcus aureus and Haemophilus influenzae was higher in CF versus PCD (p<0.05), but absolute neutrophil counts were higher in PCD patients (p=0.02). While sputum elastase activity was similar in PCD and CF at the time of exacerbation, it decreased with antibiotic therapy in PCD (p<0.05) but not CF patients.PCD patients differ from those with CF in their responses to treatment of pulmonary exacerbations, with higher neutrophil elastase activity persisting in the CF airways at the end of treatment.

scholarly journals Transcriptional and posttranscriptional modulation of human neutrophil elastase gene expression

Blood ◽  
1992 ◽  
Vol 79 (10) ◽  
pp. 2733-2740
Author(s):  
K Yoshimura ◽  
RG Crystal
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Gene Transcription ◽  
Neutrophil Elastase ◽  
Human Neutrophil ◽  
Leukemia Cell Line ◽  
Human Neutrophil Elastase ◽  
Mrna Levels ◽  
Mrna Transcript ◽  
Lineage Differentiation

Human neutrophil elastase (NE), a 29-Kd potent serine protease stored in azurophilic granules of mature neutrophils, is coded for by the NE gene, a single copy gene with 5 exons spanning a 6-kb segment of chromosome 11 at q14. With the knowledge that the NE gene expression is limited to early myeloid cell differentiation, mechanisms modulating expression of the NE gene were evaluated in the HL-60 promyelocytic leukemia cell line, a model of early bone marrow precursor cells. Consistent with the presence of NE messenger RNA (mRNA) transcripts in undifferentiated HL-60 cells, nuclear transcription run-on analyses showed that HL-60 cells actively transcribed the NE gene. However, the transcription rate of the NE gene was relatively low, only 40% of the myeloperoxidase gene, a gene expressed in parallel with NE. When induced toward the mononuclear phagocytic lineage with phorbol 12- myristate 13-acetate (PMA), HL-60 cells exhibited marked suppression of NE gene transcription, declining to 17% of the resting rate within 2 days. Induction toward mononuclear phagocytic lineage differentiation caused no change in NE mRNA transcript half-life (T1/2), but mRNA levels decreased markedly over time, with levels undetectable 1.5 days after PMA stimulation. In contrast, when induced toward the myelocytic lineage with dimethyl sulfoxide, the rate of NE gene transcription increased 1.9-fold within 5 days. Interestingly, the mRNA transcript levels increased 2.5-fold by 5 days despite the fact that induction toward myelocytic lineage differentiation was accompanied by a marked reduction of NE mRNA transcript T1/2. Together, these observations suggest that the NE gene expression during bone marrow differentiation is modulated mainly at the transcriptional level, with some posttranscriptional modulation contributing, particularly during myelocytic lineage differentiation.

Identification of Hox genes in newborn lung and effects of gestational age and retinoic acid on their expression

1994 ◽  
Vol 266 (4) ◽  
pp. L448-L454 ◽  
Author(s):  
C. W. Bogue ◽  
I. Gross ◽  
H. Vasavada ◽  
D. W. Dynia ◽  
C. M. Wilson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Retinoic Acid ◽  
Gestational Age ◽  
Hox Genes ◽  
Homeobox Genes ◽  
Mouse Lung ◽  
Newborn Mouse ◽  
Mrna Levels ◽  
Rat Lung ◽  
Hox Gene

Hox genes are sequence-specific DNA transcription factors, which are important in embryonic development and are expressed in a number of fetal tissues, including the lung. Additionally, retinoic acid (RA) has been shown to modulate Hox gene expression in a number of cell types. The specific aims of this study were to 1) identify those Hox genes expressed in newborn mouse lung using reverse transcription-polymerase chain reaction (RT-PCR), 2) study the ontogeny of Hox gene expression in fetal mouse and rat lung by Northern analysis using cDNAs for mouse Hox genes, and 3) study the effects of RA on whole lung Hox mRNA levels in cultured fetal rat lung explants. Our data show that 16 different homeobox genes are expressed in newborn mouse lung. This includes seven Hox genes not previously identified in lung, as well as the divergent homeobox gene Hex. Steady-state mRNA levels of Hox A5 (Hox 1.3), B5 (Hox 2.1), B6 (Hox 2.2), and B8 (Hox 2.4) decrease with advancing gestational age in mouse lungs (E14 to adult). Similarly, Hox A5, B5, and B6 follow the same decreasing pattern of expression with advancing gestational age in rat lungs (E15 to adult). RA treatment of E17 rat lung explants in culture resulted in a significant dose- and time-dependent increase in Hox A5, B5, and B6 mRNA levels. The highest mRNA levels were seen in explants treated with 1 x 10(-5) M RA for 4-16 h. We conclude that there are many homeobox genes expressed in developing rodent lung and that their mRNA levels are affected by both gestational age and RA.

Neutrophil elastase induces mucus cell metaplasia in mouse lung

2004 ◽  
Vol 287 (6) ◽  
pp. L1293-L1302 ◽  
Author(s):  
Judith A. Voynow ◽  
Bernard M. Fischer ◽  
David E. Malarkey ◽  
Lauranell H. Burch ◽  
Teresa Wong ◽  
...  
Keyword(s):  
Proteolytic Activity ◽  
Goblet Cell ◽  
Neutrophil Elastase ◽  
Mouse Lung ◽  
Pathological Feature ◽  
Cell Hyperplasia ◽  
Mucus Cell ◽  
Elastase Inhibitor ◽  
Cell Counts ◽  
Goblet Cell Metaplasia

Goblet cell hyperplasia in the superficial airway epithelia is a signature pathological feature of chronic bronchitis and cystic fibrosis. In these chronic inflammatory airway diseases, neutrophil elastase (NE) is found in high concentrations in the epithelial lining fluid. NE has been reported to trigger mucin secretion and increase mucin gene expression in vitro. We hypothesized that chronic NE exposure to murine airways in vivo would induce goblet cell metaplasia. Human NE (50 μg) or PBS saline was aspirated intratracheally by male Balb/c (6 wk of age) mice on days 1, 4, and 7. On days 8, 11, and 14, lung tissues for histology and bronchoalveolar lavage (BAL) samples for cell counts and cytokine levels were obtained. NE induced Muc5ac mRNA and protein expression and goblet cell metaplasia on days 8, 11, and 14. These cellular changes were the result of proteolytic activity, since the addition of an elastase inhibitor, methoxysuccinyl Ala-Ala-Pro-Val chloromethylketone (AAPV-CMK), blocked NE-induced Muc5ac expression and goblet cell metaplasia. NE significantly increased keratinocyte-derived chemokine and IL-5 in BAL and increased lung tissue inflammation and BAL leukocyte counts. The addition of AAPV-CMK reduced these measures of inflammation to control levels. These experiments suggest that NE proteolytic activity initiates an inflammatory process leading to goblet cell metaplasia.

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