Effects of topical corticosteroids on inflammatory mediator-induced eicosanoid release by human airway epithelial cells.
BACKGROUND
Airway epithelial cells are among the first cells to come in contact with aerosolized corticosteroids. However, the relative potencies and time course of action of the several commonly used aerosolized corticosteroids on eicosanoid production by airway epithelial cells are unknown.


OBJECTIVE
This study compared the effects of fluticasone, budesonide, and triamcinolone on eicosanoid output by human airway epithelial cells in vitro. We also determined the spectrum of eicosanoids affected and the mechanism for corticosteroid action.


METHODS
Cultured BEAS-2B airway epithelial cells (a transformed cell line) were exposed to corticosteroids (1 nmol/L to 1 micromol/L) for 2 to 48 hours and then assayed for basal- and bradykinin (BK)-stimulated eicosanoid output. The eicosanoid profile was identified by HPLC in tritiated arachidonic acid prelabelled cells, and PGE2, the major eicosanoid product, was quantitated by RIA. The effect of corticosteroids on the immunoreactivity of key proteins involved in eicosanoid metabolism (ie, cyclooxygenase [COX], phospholipase A2 [PLA2], and Clara cell protein, a PLA2 inhibitor) was determined by Western blotting.


RESULTS
Eicosanoid output was largely confined to prostaglandins with values of 5 +/- 2 and 82 +/- 35 ng PGE2/10(6) cells for basal- and BK stimulation, respectively (n = 8). All 3 corticosteroids inhibited basal- and BK-induced PGE2 output in a dose- and time-dependent manner. Fluticasone and budesonide completely eliminated PGE2 output in nanomolar concentrations in contrast to triamcinolone, which required micromolar concentration. The rank order of potency was: fluticasone = budesonide > triamcinolone. The time course of action for PGE2 inhibition also differed, with budesonide acting more slowly than the other 2 corticosteroids (P = .04). All 3 corticosteroids markedly reduced COX2 with little effect on COX1, cPLA2 (Type IV), or iPLA2 (Type VI) immunoreactivity or their relative distribution in cytosol versus membrane fractions. Clara cell protein immunoreactivity was undetectable in control and corticosteroid-treated cell lysates.


CONCLUSIONS
These results show that in a human airway epithelial cell line, the 3 inhaled corticosteroids commonly used to treat asthma differ in onsets of action as inhibitors of prostaglandin synthesis and vary considerably in potency. All 3 corticosteroids act mechanistically in similar fashion by inhibiting COX2 synthesis.
Aksoy
M O
MO
Pulmonary Division, Department of Medicine, Temple University School of Medicine, Philadelphia, USA.
Li
X
X
Borenstein
M
M
Yi
Y
Y
Kelsen
S G
SG
D000287:Administration, Topical; D000305:Adrenal Cortex Hormones / Q000008:administration & dosage*; D000730:Androstadienes / Q000008:administration & dosage; D000893:Anti-Inflammatory Agents / Q000008:administration & dosage; D001920:Bradykinin / Q000494:pharmacology; D001980:Bronchi / Q000166:cytology; D019819:Budesonide / Q000008:administration & dosage; D002478:Cells, Cultured; D015232:Dinoprostone / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015777:Eicosanoids / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism*; D000068298:Fluticasone; D005938:Glucocorticoids; D006801:Humans; D018836:Inflammation Mediators / Q000494:pharmacology*; D010741:Phospholipases A / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D054467:Phospholipases A2; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D014176:Protein Biosynthesis; D013997:Time Factors; D014221:Triamcinolone / Q000008:administration & dosage; D014598:Uteroglobin
D000305:Adrenal Cortex Hormones; D000730:Androstadienes; D000893:Anti-Inflammatory Agents; D015777:Eicosanoids; D004791:Enzyme Inhibitors; D005938:Glucocorticoids; D018836:Inflammation Mediators; C556389:SCGB1A1 protein, human; D014221:Triamcinolone; D019819:Budesonide; D014598:Uteroglobin; D000068298:Fluticasone; D011451:Prostaglandin-Endoperoxide Synthases; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone; D001920:Bradykinin