Abstract
Objective - To isolate and culture primary equine airway epithelial cells in vitro and elucidate the major cytokines involved in expression of the gel-forming mucin gene MUC5AC in horses. Sample Population - 12 tracheas obtained within 5 hours after euthanasia from horses free from respiratory tract disease. Procedures - Tracheal rings were digested overnight in 0.2% protease, and dissociated airway epithelial cells were grown in a serum-free defined medium at an air-liquid interface until confluence was achieved. Differentiated airway epithelial cells were treated with a panel of recombinant equine cytokines followed by quantitative reverse transcriptase PCR assay for mRNA of equine MUC5AC and the control gene glyceraldehyde 3-phosphate dehydrogenase. Cultures were incubated in the presence of isohelenin, a nuclear factor κB-DNA-binding inhibitor, to investigate transcriptional regulation of MUC5AC. Results - Light and electron microscopy revealed a differentiated epithelium with ciliated cells, nonciliated mucous cells, and basal-like cells. Recombinant equine tumor necrosis factor-α was the major mediator in the cytokine panel that significantly increased MUC5AC mRNA by a factor of 5 in a dose- and time-dependent manner. This enhancement was attenuated by isohelenin. Conclusions and Clinical Relevance - Data suggested that a nuclear factor κB-based transcriptional mechanism is involved in induction of MUC5AC expression by tumor necrosis factor-α. Understanding the molecular mechanism of cytokine-enhanced MUC5AC expression in horses may lead to better treatment options and understanding of the pathogenesis of equine pulmonary diseases.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 690-696 |
| Number of pages | 7 |
| Journal | American Journal of Veterinary Research |
| Volume | 71 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 2010 |
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ASJC Scopus subject areas
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Evaluation of MUC5AC expression and upregulation in airway epithelial cells of horses. / Oslund, Karen L.; Adamson, Grete; Wu, Reen.
In: American Journal of Veterinary Research, Vol. 71, No. 6, 06.2010, p. 690-696.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Evaluation of MUC5AC expression and upregulation in airway epithelial cells of horses
AU - Oslund, Karen L.
AU - Adamson, Grete
AU - Wu, Reen
PY - 2010/6
Y1 - 2010/6
N2 - Objective - To isolate and culture primary equine airway epithelial cells in vitro and elucidate the major cytokines involved in expression of the gel-forming mucin gene MUC5AC in horses. Sample Population - 12 tracheas obtained within 5 hours after euthanasia from horses free from respiratory tract disease. Procedures - Tracheal rings were digested overnight in 0.2% protease, and dissociated airway epithelial cells were grown in a serum-free defined medium at an air-liquid interface until confluence was achieved. Differentiated airway epithelial cells were treated with a panel of recombinant equine cytokines followed by quantitative reverse transcriptase PCR assay for mRNA of equine MUC5AC and the control gene glyceraldehyde 3-phosphate dehydrogenase. Cultures were incubated in the presence of isohelenin, a nuclear factor κB-DNA-binding inhibitor, to investigate transcriptional regulation of MUC5AC. Results - Light and electron microscopy revealed a differentiated epithelium with ciliated cells, nonciliated mucous cells, and basal-like cells. Recombinant equine tumor necrosis factor-α was the major mediator in the cytokine panel that significantly increased MUC5AC mRNA by a factor of 5 in a dose- and time-dependent manner. This enhancement was attenuated by isohelenin. Conclusions and Clinical Relevance - Data suggested that a nuclear factor κB-based transcriptional mechanism is involved in induction of MUC5AC expression by tumor necrosis factor-α. Understanding the molecular mechanism of cytokine-enhanced MUC5AC expression in horses may lead to better treatment options and understanding of the pathogenesis of equine pulmonary diseases.
AB - Objective - To isolate and culture primary equine airway epithelial cells in vitro and elucidate the major cytokines involved in expression of the gel-forming mucin gene MUC5AC in horses. Sample Population - 12 tracheas obtained within 5 hours after euthanasia from horses free from respiratory tract disease. Procedures - Tracheal rings were digested overnight in 0.2% protease, and dissociated airway epithelial cells were grown in a serum-free defined medium at an air-liquid interface until confluence was achieved. Differentiated airway epithelial cells were treated with a panel of recombinant equine cytokines followed by quantitative reverse transcriptase PCR assay for mRNA of equine MUC5AC and the control gene glyceraldehyde 3-phosphate dehydrogenase. Cultures were incubated in the presence of isohelenin, a nuclear factor κB-DNA-binding inhibitor, to investigate transcriptional regulation of MUC5AC. Results - Light and electron microscopy revealed a differentiated epithelium with ciliated cells, nonciliated mucous cells, and basal-like cells. Recombinant equine tumor necrosis factor-α was the major mediator in the cytokine panel that significantly increased MUC5AC mRNA by a factor of 5 in a dose- and time-dependent manner. This enhancement was attenuated by isohelenin. Conclusions and Clinical Relevance - Data suggested that a nuclear factor κB-based transcriptional mechanism is involved in induction of MUC5AC expression by tumor necrosis factor-α. Understanding the molecular mechanism of cytokine-enhanced MUC5AC expression in horses may lead to better treatment options and understanding of the pathogenesis of equine pulmonary diseases.
UR - http://www.scopus.com/inward/record.url?scp=77954042787&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=77954042787&partnerID=8YFLogxK
U2 - 10.2460/ajvr.71.6.690
DO - 10.2460/ajvr.71.6.690
M3 - Article
C2 - 20513186
AN - SCOPUS:77954042787
VL - 71
SP - 690
EP - 696
JO - American Journal of Veterinary Research
JF - American Journal of Veterinary Research
SN - 0002-9645
IS - 6
ER -