Inhibition of VCAM-1 expression in human bronchial epithelial cells by glucocorticoids

Jun Atsuta, Jim Plitt, Bruce S. Bochner, Robert P. Schleimer

Research output: Contribution to journalArticle

Abstract

We have demonstrated previously that cytokines induce surface expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) on BEAS-2B bronchial epithelial cells in vitro. The present studies demonstrate glucocorticoid inhibition of cytokine-induced VCAM-1 expression as detected using flow cytometry and Northern blot analysis. Several commonly used inhaled glucocorticoids were tested for their ability to inhibit VCAM-1 and ICAM-1 expression. All glucocorticoids tested inhibited VCAM-1 expression in a dose-dependent manner. No inhibition of ICAM-1 expression was observed. The most potent of the glucocorticoids tested for inhibition of VCAM-1 expression were mometasone furoate and fluticasone propionate (FP), which had IC50 values (i.e., concentrations at which each glucocorticoid produced 50% inhibition) of under 10 pM. Budesonide, triamcinolone acetonide, and beclomethasone dipropionate (BDP) had intermediate potency, and hydrocortisone and the BDP metabolite beclomethasone-17-monopropionate were the least potent of the steroids tested. Kinetic analysis of the ability of FP to inhibit VCAM-1 expression revealed that preincubation with FP for 3 h completely inhibited VCAM-1 expression induced by tumor necrosis factor-α (TNF-α). FP inhibited VCAM-1 expression by 50% even when added as late as 6 h after stimulation with TNF-α. Using Northern blot analysis, we confirmed inhibition of VCAM-1 and ICAM-1 messenger RNA (mRNA) expression by FP. Pretreatment with FP (10 11 M to about 10 7 M, 24 h) inhibited TNF-α-induced VCAM-1 mRNA expression in BEAS-2B in a dose-dependent manner, but did not inhibit expression of ICAM-1 mRNA. Studies with actinomycin D indicate that FP treatment accelerated the degradation of TNF-α-induced VCAM-1 mRNA. FP (10-7 M) also inhibited VCAM-1 mRNA expression induced by TNF-α in primary human bronchial epithelial cells as assessed by reverse transcription-polymerase chain reaction. These results suggest that suppression of epithelial VCAM-1 expression by glucocorticoids may contribute to their anti-inflammatory effects.

Original languageEnglish (US)
Pages (from-to)643-650
Number of pages8
JournalAmerican Journal of Respiratory Cell and Molecular Biology
Volume20
Issue number4
StatePublished - 1999

Fingerprint

Vascular Cell Adhesion Molecule-1
Glucocorticoids
Epithelial Cells
Intercellular Adhesion Molecule-1
Tumor Necrosis Factor-alpha
Messenger RNA
Beclomethasone
Mometasone Furoate
Northern Blotting
Cytokines
Triamcinolone Acetonide
Budesonide
Flow cytometry
Fluticasone
Polymerase chain reaction
Dactinomycin
Transcription
Metabolites
Inhibitory Concentration 50
Reverse Transcription

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Pulmonary and Respiratory Medicine

Cite this

Inhibition of VCAM-1 expression in human bronchial epithelial cells by glucocorticoids. / Atsuta, Jun; Plitt, Jim; Bochner, Bruce S.; Schleimer, Robert P.

In: American Journal of Respiratory Cell and Molecular Biology, Vol. 20, No. 4, 1999, p. 643-650.

Research output: Contribution to journalArticle

Atsuta, Jun ; Plitt, Jim ; Bochner, Bruce S. ; Schleimer, Robert P. / Inhibition of VCAM-1 expression in human bronchial epithelial cells by glucocorticoids. In: American Journal of Respiratory Cell and Molecular Biology. 1999 ; Vol. 20, No. 4. pp. 643-650.
@article{9e15e69e3b45466287da7d8b82b3e8e0,
title = "Inhibition of VCAM-1 expression in human bronchial epithelial cells by glucocorticoids",
abstract = "We have demonstrated previously that cytokines induce surface expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) on BEAS-2B bronchial epithelial cells in vitro. The present studies demonstrate glucocorticoid inhibition of cytokine-induced VCAM-1 expression as detected using flow cytometry and Northern blot analysis. Several commonly used inhaled glucocorticoids were tested for their ability to inhibit VCAM-1 and ICAM-1 expression. All glucocorticoids tested inhibited VCAM-1 expression in a dose-dependent manner. No inhibition of ICAM-1 expression was observed. The most potent of the glucocorticoids tested for inhibition of VCAM-1 expression were mometasone furoate and fluticasone propionate (FP), which had IC50 values (i.e., concentrations at which each glucocorticoid produced 50{\%} inhibition) of under 10 pM. Budesonide, triamcinolone acetonide, and beclomethasone dipropionate (BDP) had intermediate potency, and hydrocortisone and the BDP metabolite beclomethasone-17-monopropionate were the least potent of the steroids tested. Kinetic analysis of the ability of FP to inhibit VCAM-1 expression revealed that preincubation with FP for 3 h completely inhibited VCAM-1 expression induced by tumor necrosis factor-α (TNF-α). FP inhibited VCAM-1 expression by 50{\%} even when added as late as 6 h after stimulation with TNF-α. Using Northern blot analysis, we confirmed inhibition of VCAM-1 and ICAM-1 messenger RNA (mRNA) expression by FP. Pretreatment with FP (10 11 M to about 10 7 M, 24 h) inhibited TNF-α-induced VCAM-1 mRNA expression in BEAS-2B in a dose-dependent manner, but did not inhibit expression of ICAM-1 mRNA. Studies with actinomycin D indicate that FP treatment accelerated the degradation of TNF-α-induced VCAM-1 mRNA. FP (10-7 M) also inhibited VCAM-1 mRNA expression induced by TNF-α in primary human bronchial epithelial cells as assessed by reverse transcription-polymerase chain reaction. These results suggest that suppression of epithelial VCAM-1 expression by glucocorticoids may contribute to their anti-inflammatory effects.",
author = "Jun Atsuta and Jim Plitt and Bochner, {Bruce S.} and Schleimer, {Robert P.}",
year = "1999",
language = "English (US)",
volume = "20",
pages = "643--650",
journal = "American Journal of Respiratory Cell and Molecular Biology",
issn = "1044-1549",
publisher = "American Thoracic Society",
number = "4",

}

TY - JOUR

T1 - Inhibition of VCAM-1 expression in human bronchial epithelial cells by glucocorticoids

AU - Atsuta, Jun

AU - Plitt, Jim

AU - Bochner, Bruce S.

AU - Schleimer, Robert P.

PY - 1999

Y1 - 1999

N2 - We have demonstrated previously that cytokines induce surface expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) on BEAS-2B bronchial epithelial cells in vitro. The present studies demonstrate glucocorticoid inhibition of cytokine-induced VCAM-1 expression as detected using flow cytometry and Northern blot analysis. Several commonly used inhaled glucocorticoids were tested for their ability to inhibit VCAM-1 and ICAM-1 expression. All glucocorticoids tested inhibited VCAM-1 expression in a dose-dependent manner. No inhibition of ICAM-1 expression was observed. The most potent of the glucocorticoids tested for inhibition of VCAM-1 expression were mometasone furoate and fluticasone propionate (FP), which had IC50 values (i.e., concentrations at which each glucocorticoid produced 50% inhibition) of under 10 pM. Budesonide, triamcinolone acetonide, and beclomethasone dipropionate (BDP) had intermediate potency, and hydrocortisone and the BDP metabolite beclomethasone-17-monopropionate were the least potent of the steroids tested. Kinetic analysis of the ability of FP to inhibit VCAM-1 expression revealed that preincubation with FP for 3 h completely inhibited VCAM-1 expression induced by tumor necrosis factor-α (TNF-α). FP inhibited VCAM-1 expression by 50% even when added as late as 6 h after stimulation with TNF-α. Using Northern blot analysis, we confirmed inhibition of VCAM-1 and ICAM-1 messenger RNA (mRNA) expression by FP. Pretreatment with FP (10 11 M to about 10 7 M, 24 h) inhibited TNF-α-induced VCAM-1 mRNA expression in BEAS-2B in a dose-dependent manner, but did not inhibit expression of ICAM-1 mRNA. Studies with actinomycin D indicate that FP treatment accelerated the degradation of TNF-α-induced VCAM-1 mRNA. FP (10-7 M) also inhibited VCAM-1 mRNA expression induced by TNF-α in primary human bronchial epithelial cells as assessed by reverse transcription-polymerase chain reaction. These results suggest that suppression of epithelial VCAM-1 expression by glucocorticoids may contribute to their anti-inflammatory effects.

AB - We have demonstrated previously that cytokines induce surface expression of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) on BEAS-2B bronchial epithelial cells in vitro. The present studies demonstrate glucocorticoid inhibition of cytokine-induced VCAM-1 expression as detected using flow cytometry and Northern blot analysis. Several commonly used inhaled glucocorticoids were tested for their ability to inhibit VCAM-1 and ICAM-1 expression. All glucocorticoids tested inhibited VCAM-1 expression in a dose-dependent manner. No inhibition of ICAM-1 expression was observed. The most potent of the glucocorticoids tested for inhibition of VCAM-1 expression were mometasone furoate and fluticasone propionate (FP), which had IC50 values (i.e., concentrations at which each glucocorticoid produced 50% inhibition) of under 10 pM. Budesonide, triamcinolone acetonide, and beclomethasone dipropionate (BDP) had intermediate potency, and hydrocortisone and the BDP metabolite beclomethasone-17-monopropionate were the least potent of the steroids tested. Kinetic analysis of the ability of FP to inhibit VCAM-1 expression revealed that preincubation with FP for 3 h completely inhibited VCAM-1 expression induced by tumor necrosis factor-α (TNF-α). FP inhibited VCAM-1 expression by 50% even when added as late as 6 h after stimulation with TNF-α. Using Northern blot analysis, we confirmed inhibition of VCAM-1 and ICAM-1 messenger RNA (mRNA) expression by FP. Pretreatment with FP (10 11 M to about 10 7 M, 24 h) inhibited TNF-α-induced VCAM-1 mRNA expression in BEAS-2B in a dose-dependent manner, but did not inhibit expression of ICAM-1 mRNA. Studies with actinomycin D indicate that FP treatment accelerated the degradation of TNF-α-induced VCAM-1 mRNA. FP (10-7 M) also inhibited VCAM-1 mRNA expression induced by TNF-α in primary human bronchial epithelial cells as assessed by reverse transcription-polymerase chain reaction. These results suggest that suppression of epithelial VCAM-1 expression by glucocorticoids may contribute to their anti-inflammatory effects.

UR - http://www.scopus.com/inward/record.url?scp=0033111565&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033111565&partnerID=8YFLogxK

M3 - Article

C2 - 10100995

AN - SCOPUS:0033111565

VL - 20

SP - 643

EP - 650

JO - American Journal of Respiratory Cell and Molecular Biology

JF - American Journal of Respiratory Cell and Molecular Biology

SN - 1044-1549

IS - 4

ER -