Document Detail


Interleukin-4 enhances 15-lipoxygenase activity and incorporation of 15(S)-HETE into cellular phospholipids in cultured pulmonary epithelial cells.
MedLine Citation:
PMID:  9870918     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
15(S)-Hydroxyeicosatetraenoic acid (15[S]-HETE) is a 15-lipoxygenase (15-LO) metabolite that may play an important role in different pulmonary diseases. 15-HETE is synthesized by different epithelial cells and may be subsequently incorporated into cellular phospholipids. We studied the role of interleukin-4 (IL-4) on 15-LO activity and on 15(S)-HETE incorporation into cellular phospholipids by WI-26 pulmonary epithelial cells. 15-LO activity was evaluated by measuring 15(S)-HETE production, through combined reverse-phase-high-pressure liquid chromatography (RP-HPLC) separation and specific radioimmunoassay (RIA), after incubation with arachidonic acid (AA). We also studied 15-LO messenger RNA (mRNA) expression, using primed in situ (PRINS) labeling. IL-4 (10 ng/ml) markedly increased the percentage of 15-LO mRNA-bearing cells as well as 15-LO activity after 24, 48, and 72 h, with a maximal response at 48 h. Uptake and incorporation into cellular phospholipid was studied with [3H]15(S)-HETE, which showed that IL-4 was able to increase significantly 15(S)-HETE incorporation into WI-26 cells, with a maximal effect observed at 72 h. Cellular-lipid-associated [3H]15(S)-HETE, evaluated with RP-HPLC after base-catalyzed hydrolysis, increased concomitantly with disappearance of the radiolabel from the supernatant. Class separation of cellular lipids with normal-phase HPLC (NP-HPLC) showed that IL-4 increased [3H]15(S)- HETE incorporation mainly in the phosphatidylinositol (PI) fraction. The ability of IL-4 to promote 15-LO activity and incorporation into cellular phospholipids of human lung epithelial cells may be important in airway inflammation and in modulation of the potential autocrine function of 15(S)-HETE.
Authors:
M Profita; A M Vignola; A Sala; A Mirabella; L Siena; E Pace; G Folco; G Bonsignore
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  American journal of respiratory cell and molecular biology     Volume:  20     ISSN:  1044-1549     ISO Abbreviation:  Am. J. Respir. Cell Mol. Biol.     Publication Date:  1999 Jan 
Date Detail:
Created Date:  1999-02-04     Completed Date:  1999-02-04     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  8917225     Medline TA:  Am J Respir Cell Mol Biol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  61-8     Citation Subset:  IM    
Affiliation:
Istituto di Fisiopatologia Respiratoria, Consiglio Nazionale delle Ricerche, Italy.
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MeSH Terms
Descriptor/Qualifier:
Arachidonate 15-Lipoxygenase / genetics,  metabolism*
Cell Line
Epithelial Cells / metabolism
Gene Expression
Humans
Hydroxyeicosatetraenoic Acids / metabolism*
Interleukin-4 / pharmacology*
Lung / metabolism*
Phospholipids / metabolism*
RNA, Messenger / metabolism
Tritium
Chemical
Reg. No./Substance:
0/Hydroxyeicosatetraenoic Acids; 0/Phospholipids; 0/RNA, Messenger; 10028-17-8/Tritium; 207137-56-2/Interleukin-4; 73945-47-8/15-hydroxy-5,8,11,13-eicosatetraenoic acid; EC 1.13.11.33/Arachidonate 15-Lipoxygenase

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