The effects of glucocorticoids on phorbol ester and cytokine stimulated transcription factor activation in human lung

doi:10.1016/0024-3205(94)00243-6

Life Sciences

Volume 55, Issue 14, 1994, Pages 1147-1153

https://doi.org/10.1016/0024-3205(94)00243-6 Get rights and content

Abstract

Glucocorticoids have a wide variety of effects which result in the dampening of inflammatory and immune responses and other challenges to homeostasis. An important site of steroid action may be on the control of transcription factor binding to DNA. The interaction of the transcription factors, activator protein 1 (AP-1) and nuclear factor kappa from B cells (NFκB) with DNA and glucocorticoid receptors was analysed by gel mobility shift assays following stimulation by tumour necrosis factor alpha (TNFα) and a phorbol ester (PMA) that activates protein kinase C. PMA and TNFα both caused significant (180–340%) increases in AP-1 and NFκB DNA binding which peaked at 15 minutes and decreased to a constant elevated level at between 1–3 hrs and was sustained for 24hrs. Dexamethasone (1μM) caused a rapid and long lasting 40–50% decrease in both AP-1 and NFκB DNA binding lasting over 24 hrs. Combined treatment with dexamethasone and PMA or TNFα prevented the increase in both AP-1 and NFκB binding due to PMA and TNFα returning levels to those seen in control untreated samples. This suggests that in human lung, the glucocorticoid receptor functionally interacts within the nucleus with other transcription factors that are induced by inflammatory mediators such as cytokines. This may be an important molecular site of steroid action in chronic inflammatory lung diseases such as asthma.

References (21)

R. Schüle et al.
Cell
(1990)
C. Jonat et al.
Cell
(1990)
H-F. Yang-Yen et al.
Cell
(1990)
K.J. Tracey et al.
Lancet
(1989)
M.J. Lenardo et al.
Cell
(1989)
M.E. Jackson
J. Cell Sci.
(1991)
A. Munck et al.
Am. Rev. Respir. Dis.
(1990)
P.J. Barnes
N. Engl. J. Med.
(1989)
D.S. Latchman
Biochem. J.
(1990)
J. Kelley
Am. Rev. Respir. Dis.
(1990)

There are more references available in the full text version of this article.

Cited by (75)

Peripheral blood mononuclear cell NF-κB p105 mRNA decreases during asthmatic attacks
2008, Biomedicine and Pharmacotherapy
Citation Excerpt :
Within resting cells, NF-κB is retained in the cytoplasm, complexed to an inhibitor protein from the inhibitor of κB (IκB) family. NF-κB is cleaved and released by various activation signals such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) [6,7], interleukin-2 (IL-2), leukotriene B4 (LTB4), allergens [8–10], mitogens, lipopolysaccharide, viral infection [11–13], oxidative stress [14], and exposure to reactive oxygen [15]. Stimulation activates the IκB kinase (IKK) complex, which then phosphorylates IκB.
NF-κB is a transcription factor involved in expression of many inflammatory cytokines, chemical transmitters, and adhesion molecules. It has been reported to play a major role in the pathogenesis of asthma. NF-κB p50, which is the actual subunit that results from the cleavage of p105, is required for the induction of eosinophilia via IL-5 and chemokines.
The subjects were 10 patients with a mean age of 59.3 years (14–82 years). NF-κB p105 mRNA in peripheral blood mononuclear cells during the presence or absence of asthmatic attacks was investigated. Total RNA was extracted from peripheral blood mononuclear cells. After cDNA was synthesized using random primers, NF-κB p105 mRNA level was measured by real-time polymerase chain reaction.
The NF-κB p105 mRNA level in peripheral blood mononuclear cells was lower during asthmatic attacks than in the absence of attacks, showing a significant difference (Wilcoxon's signed rank test: p < 0.01).
A drop in NF-κB p105 during an asthma attack could result in increased NF-κB activity. There is a possibility that a change in the NF-κB p105 mRNA level might indicate some pathogenetic state in bronchial asthma attacks.
Anti-inflammatory effects of hydroxycinnamic acid derivatives
2007, Biochemical and Biophysical Research Communications
Citation Excerpt :
Nitrate production was suppressed by CAF, and SOD activity was enhanced by CAF. NF-κB activation has been described in a variety of inflammatory disease models [21–23] and is thought to significantly contribute to the progression of the disease through the enhanced expression of target inflammatory genes. ROS would be involved in activation of the IKK complex, inducing phosphorylation of IκB and activation of NF-κB [24–26].
NF-κB family of transcription factors are involved in numerous cellular processes, including differentiation, proliferation, and inflammation. It was reported that hydroxycinnamic acid derivatives (HADs) are inhibitors of NF-κB activation. Rice bran oil contains a lot of phytosteryl ferulates, one of HADs. We have investigated effects of phytosteryl ferulates on NF-κB activation in macrophage. Cycloartenyl ferulate (CAF), one of phytosteryl ferulates, significantly reduced lipopolysaccharide (LPS)-induced NO production and mRNA expression of inducible NO synthase and cyclooxygenese-2 but upregulated SOD activity. Electrophoresis mobility shift assay revealed that CAF inhibited DNA-binding of NF-κB. CAF and phytosteryl ferulates probably have potentially anti-inflammatory properties.
Tumor necrosis factor-α: Molecular and cellular mechanisms in skeletal pathology
2003, Gene
Tumor necrosis factor-α (TNF) is one member of a large family of inflammatory cytokines that share common signal pathways, including activation of the transcription factor nuclear factor kappa B (Nf-κB) and stimulation of the apoptotic pathway. Data derived from early work supported a role for TNF as a skeletal catabolic agent that stimulates osteoclastogenesis while simultaneously inhibiting osteoblast function. The finding that estrogen deficiency was associated with increased production of cytokines led to a barrage of studies and lively debate on the relative contributions of TNF and other cytokines on bone loss, on the potential cell sources of TNF in the bone microenvironment, and on the mechanism of TNF action. TNF has a central role in bone pathophysiology. TNF is necessary for stimulation of osteoclastogenesis along with the receptor activator of Nf-κB ligand (RANKL). TNF also stimulates osteoblasts in a manner that hinders their bone-formative action. TNF suppresses recruitment of osteoblasts from progenitor cells, inhibits the expression of matrix protein genes, and stimulates expression of genes that amplify osteoclastogenesis. TNF may also affect skeletal metabolism by inducing resistance to 1,25-dihydroxyvitamin D₃ (1,25(OH)₂D₃) by a mechanism that extends to other members of the steroid hormone nuclear receptor family. Thus, TNF assails bone at many levels. This review will focus on the cellular and molecular mechanisms of TNF action in the skeleton that result in increased bone resorption and impaired formation. TNF and its signal pathway remains an important target for the development of new therapies for bone loss from osteoporosis and inflammatory arthritis.
High Dose of Inhaled Fluticasone Reduces High Levels of Urinary Leukotriene E<inf>4</inf> in the Early Morning in Mild and Moderate Nocturnal Asthma
2003, Chest
Citation Excerpt :
We could not determine the mechanism by which ICS suppressed LT production. Hancox et al30 reported that 6 weeks of treatment with inhaled budesonide (400 μg/d) reduced the binding activity of the proinflammatory transcription factor nuclear factor-κB (NFkB) in bronchial mucosa in asthmatic subjects, which supports the finding that corticosteroids reduce the level of NFkB binding DNA in human lung tissue and peripheral blood mononuclear cells.3132 Wilson et al33 also demonstrated that the anti-inflammatory effects of ICS are mediated through inhibition of NFkB-regulated gene expression.
The circadian variation in urinary leukotriene E₄ (LTE₄) excretion with a morning acrophase has recently been reported in nocturnal asthma (NA); however, the effects of inhaled corticosteroids (ICS) on this circadian rhythmicity of leukotriene (LT) in patients with NA are controversial.
We first measured peak expiratory flow (PEF), urinary LTE₄, 11-dehydro-thromboxane B₂ (TXB₂), and creatinine levels six times every 4 h for 24 h in two groups: patients with mild-to-moderate, steroid-naive NA (n = 10, group A), and patients with severe NA treated with high-dose ICS (n = 10, group B). Next, group A patients received 2 weeks of treatment with 800 μg/d of inhaled fluticasone propionate (FP), and we compared the measured parameters before and after treatment.
In group A, a circadian rhythm in urinary LTE₄ with peak levels at approximately 4 am associated with reduced PEF was observed. Group B had suppression of urinary LTE₄ excretion and had no circadian rhythmicity, as seen in group A, despite a dip in PEF at 4 am. A high dose of FP in group A significantly (p < 0.05) reduced LTE₄ levels and abolished the circadian rhythm, as well as improving PEF. We found no significant difference in the circadian rhythm of urinary 11-dehydro-TXB₂ between groups A and B, and high-dose FP partially decreased urinary 11-dehydro-TXB₂ levels but not significantly.
A high-dose of ICS reduced urinary LTE₄ levels and abolished their circadian variation in patients with asthma, suggesting that LT might contribute to the mechanism of NA.
Refractory asthma
2002, Medical Clinics of North America
Review of the molecular and cellular mechanisms of action of glucocorticoids for use in asthma
2002, Pulmonary Pharmacology and Therapeutics
Asthma is characterized by inflammation in the lung and glucocorticoids (GCs) are the most clinically effective treatment available. The success of chronic GC therapy for asthma stems largely from the ability of the GC–GC receptor (GR) complex to alter transcription of a wide array of molecules involved in the inflammatory process. Many of the adverse effects of elevated systemic GC levels have been reduced through the use of inhalation as a method of administration, as opposed to oral GC. GCs exert their effects by binding to the wild-type GR, GR_α. The GR_α complex can directly or indirectly alter gene transcription by binding to specific DNA sites or by activating transcription factors. There is also evidence to support GR_α involvement in post-translational activities. In the management of asthma, the GR_α down-regulates proinflammatory mediators such as interleukin-(IL)-1, 3, and 5, and up-regulates anti-inflammatory mediators such as IκB [inhibitory molecule for nuclear factor κB1 IL-10, and 12. Newer GCs are being designed to increase potency and topical activity. Mometasone furoate (MF), has recently been developed for the treatment of asthma and inhibits key anti-inflammatory processes with a potency equal to or greater than that of fluticasone propionate. A better understanding of the molecular mechanisms involved might provide strategies for optimizing the effectiveness of GC in the treatment of asthma.

View all citing articles on Scopus

View full text

The effects of glucocorticoids on phorbol ester and cytokine stimulated transcription factor activation in human lung

Abstract

Cell

Cell

Cell

Lancet

Cell

J. Cell Sci.

Am. Rev. Respir. Dis.

N. Engl. J. Med.

Biochem. J.

Am. Rev. Respir. Dis.