CoChat
Sign Up
Cited by 197Wound Healing and TreatmentsAdrenal Hormones and DisordersInflammatory mediators and NSAID effects

The DOI matches its Crossref record; 67 of the 67 references it cites were found in a registry or an index.

Cortisol Synthesis in Epidermis Is Induced by IL-1 and Tissue Injury

Open access
Vukelic, Sasa, Stojadinovic, Olivera, Pastar, Irena, Rabach, Morgan, Krzyzanowska, Agata, Lebrun, Elizabeth, Davis, Stephen C., Resnik, Sydney, Brem, Harold, Tomic-Canic, Marjana·2011·Journal of Biological Chemistry
DOI 10.1074/jbc.m110.188268PMC PMC3060481
Open this paper in CoChat — free accountOpen PDF

Abstract

Glucocorticoids (GCs) are known inhibitors of wound healing. In this study we report the novel finding that both keratinocytes in vitro and epidermis in vivo synthesize cortisol and how this synthesis regulates wound healing. We show that epidermis expresses enzymes essential for cortisol synthesis, including steroid 11 β-hydroxylase (CYP11B1), and an enzyme that controls negative feedback mechanism, 11β-hydroxysteroid dehydrogenase 2 (11βHSD2). We also found that cortisol synthesis in keratinocytes and skin can be stimulated by ACTH and inhibited by metyrapone (CYP11B1 enzyme inhibitor). Interestingly, IL-1β, the first epidermal signal of tissue injury, induces the expression of CYP11B1 and increases cortisol production by keratinocytes. Additionally, we found induction of CYP11B1 increased production of cortisol and activation of GR pathway during wound healing ex vivo and in vivo using human and porcine wound models, respectively. Conversely, inhibition of cortisol synthesis during wound healing increases IL-1β production, suggesting that cortisol synthesis in epidermis may serve as a local negative feedback to proinflammatory cytokines. Local GCs synthesis, therefore, may provide control of the initial proinflammatory response, preventing excessive inflammation upon tissue injury. Inhibition of GC synthesis accelerated wound closure in vivo, providing the evidence that modulation of cortisol synthesis in epidermis may be an important regulatory mechanism during wound healing.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1074/jbc.m110.188268

Cites

67 of 67 references verified0 citation errors · 0 not checkable

Statistics

Figures

FIGURE 1 GRreceptor is activated in human epidermis. A , immunofluorescence staining with GR- phosphorylated Ser 211 antibody of human skin explants maintainedintheabsenceofGCsfor24hrevealedthenuclearpresenceofthereceptorinkeratinocytes.Dexamethasone( DEX )-treated skin, a positive control, confirmed nuclear translocation of GR after topical corticosteroid treatment. No staining is found in the absence of primary antibody. B , qRT- PCR confirms expression of CYP11B1 in HEK and skin. Expression levels were normalized to GAPDH. C , immunoperoxidase staining of human skin with CYP11B antibody ( Ab ) showsincreased staining of basal and first suprabasal keratinocytes, whereas staining intensity decreases in upper epidermal layers. No staining was observed in sections incubated without primary antibody. As expected, signal was present in zona fasciculata of mouse adrenal gland. D , Western blot with CYP11B1 antibodyandproteinextractsfromadrenalgland,skin,andHEKandlymphocytes( Lym ) is shown. Both keratinocytes and skin showed significant protein levels of CYP11B1 in comparison to adrenal gland (positive control), whereas no CYP11B1 was detected in lymphocytes, which do not express CYP11B1 (52) and served as negative control. E , ACTH stimulates CYP11B1 expression in keratinocytes. HEK were incubated with/without ACTH (10 -7 M) for 24 h ( n = 4). Quantitative real time PCR for the expression of CYP11B1 is shown. Expression levels were normalized to HPRT1. Error bars represent S.D. F , ACTH stimulates CYP11B1 expression in skin. Skin specimens were incubated with/without ACTH (10 -7 M) for 24 h ( n = 4). Quantitative real time PCR for the expression of CYP11B1 is shown. Error bars represent S.D.
FIGURE 1 GRreceptor is activated in human epidermis. A , immunofluorescence staining with GR- phosphorylated Ser 211 antibody of human skin explants maintainedintheabsenceofGCsfor24hrevealedthenuclearpresenceofthereceptorinkeratinocytes.Dexamethasone( DEX )-treated skin, a positive control, confirmed nuclear translocation of GR after topical corticosteroid treatment. No staining is found in the absence of primary antibody. B , qRT- PCR confirms expression of CYP11B1 in HEK and skin. Expression levels were normalized to GAPDH. C , immunoperoxidase staining of human skin with CYP11B antibody ( Ab ) showsincreased staining of basal and first suprabasal keratinocytes, whereas staining intensity decreases in upper epidermal layers. No staining was observed in sections incubated without primary antibody. As expected, signal was present in zona fasciculata of mouse adrenal gland. D , Western blot with CYP11B1 antibodyandproteinextractsfromadrenalgland,skin,andHEKandlymphocytes( Lym ) is shown. Both keratinocytes and skin showed significant protein levels of CYP11B1 in comparison to adrenal gland (positive control), whereas no CYP11B1 was detected in lymphocytes, which do not express CYP11B1 (52) and served as negative control. E , ACTH stimulates CYP11B1 expression in keratinocytes. HEK were incubated with/without ACTH (10 -7 M) for 24 h ( n = 4). Quantitative real time PCR for the expression of CYP11B1 is shown. Expression levels were normalized to HPRT1. Error bars represent S.D. F , ACTH stimulates CYP11B1 expression in skin. Skin specimens were incubated with/without ACTH (10 -7 M) for 24 h ( n = 4). Quantitative real time PCR for the expression of CYP11B1 is shown. Error bars represent S.D.CC-BY · From the PDF
Image 2
CC-BY · From the PDF
Image 3
CC-BY · From the PDF
Image 4
CC-BY · From the PDF
Image 5
CC-BY · From the PDF
FIGURE 3 IL-1 𝛃 and IGF-1 modulate CYP11B1 expression and cortisol production in HEK. A , IL-1 and ACTH stimulate cortisol production in HEK ( p < 0.01), whereas IGF-1 inhibits it. Cells were incubated with IL-1 𝛃 (10 ng/ml), IGF-1 (100 ng/ml), or ACTH (10 -7 M)for24h( n = 5). Data represent the mean ± S.D. and were analyzed by one-way ANOVA (F = 75.32; p < 0.0001) and Dunnett's post hoc test (*, p < 0.01). B , IL-1 stimulates CYP11B1 expression in HEK, and IGF-1 inhibits it. Cells were incubated with IL-1 𝛃 (10 ng/ml), IGF-1(100 ng/ml), or ACTH (10 -7 M) for 24 h ( n = 3). Quantitative real time PCR for the expression of CYP11B1 is shown. Expression levels were normalized to HPRT1. Error bars represent S.D.
FIGURE 3 IL-1 𝛃 and IGF-1 modulate CYP11B1 expression and cortisol production in HEK. A , IL-1 and ACTH stimulate cortisol production in HEK ( p < 0.01), whereas IGF-1 inhibits it. Cells were incubated with IL-1 𝛃 (10 ng/ml), IGF-1 (100 ng/ml), or ACTH (10 -7 M)for24h( n = 5). Data represent the mean ± S.D. and were analyzed by one-way ANOVA (F = 75.32; p < 0.0001) and Dunnett's post hoc test (*, p < 0.01). B , IL-1 stimulates CYP11B1 expression in HEK, and IGF-1 inhibits it. Cells were incubated with IL-1 𝛃 (10 ng/ml), IGF-1(100 ng/ml), or ACTH (10 -7 M) for 24 h ( n = 3). Quantitative real time PCR for the expression of CYP11B1 is shown. Expression levels were normalized to HPRT1. Error bars represent S.D.CC-BY · From the PDF
Image 7
CC-BY · From the PDF
Image 8
CC-BY · From the PDF
Image 9
CC-BY · From the PDF
Image 10
CC-BY · From the PDF
Image 11
CC-BY · From the PDF
Image 12
CC-BY · From the PDF
FIGURE 5 Expression of CYP11B1 and cortisol synthesis during acute wound healing in vivo . A , CYP11B1 gene expression determined using real-time qPCR showed induction at 48 h post wounding as compared with normal porcine skin ( n = 3). Error bars represent S.D. B , laser microdissected epidermal keratinocytes demonstrate induction of CYP11B1 enzyme 48 h post wounding. At the later time points CYP11B1 expression decreased back to the level of normal, unwounded skin ( n = 3). Error bars represent S.D. C , porcine wound explants show the highest levels of cortisol production 48 h after the wounding ( n = 2). Cortisol production is inhibited with metyrapone (10 -3 M) and induced by ACTH (10 -7 M).
FIGURE 5 Expression of CYP11B1 and cortisol synthesis during acute wound healing in vivo . A , CYP11B1 gene expression determined using real-time qPCR showed induction at 48 h post wounding as compared with normal porcine skin ( n = 3). Error bars represent S.D. B , laser microdissected epidermal keratinocytes demonstrate induction of CYP11B1 enzyme 48 h post wounding. At the later time points CYP11B1 expression decreased back to the level of normal, unwounded skin ( n = 3). Error bars represent S.D. C , porcine wound explants show the highest levels of cortisol production 48 h after the wounding ( n = 2). Cortisol production is inhibited with metyrapone (10 -3 M) and induced by ACTH (10 -7 M).CC-BY · From the PDF
Image 14
CC-BY · From the PDF
FIGURE6 11 𝛃 -HSD2(negativefeedbackenzyme)isinducedbyglucocorticoids. A , expression of 11 𝛃 -HSD2 during ex vivo human acute wound healing ( n = 3) is shown. Induction of 11 𝛃 -HSD2 48 h post wounding is shown. Quantitative real time PCR for the expression of 11 𝛃 -HSD2 is shown. Expression levels were normalized to HPRT1. Error bars represent S.D. B , dexamethasone( Dex , 10 -3 M)treatmentfor6hofhumanskinleadstoa4-foldinduction of 11 𝛃 -HSD2 ( n = 3). Quantitative real time PCR for the expression of 11 𝛃 -HSD2is shown. Expression levels were normalized to HPRT1. Error bars represent S.D.
FIGURE6 11 𝛃 -HSD2(negativefeedbackenzyme)isinducedbyglucocorticoids. A , expression of 11 𝛃 -HSD2 during ex vivo human acute wound healing ( n = 3) is shown. Induction of 11 𝛃 -HSD2 48 h post wounding is shown. Quantitative real time PCR for the expression of 11 𝛃 -HSD2 is shown. Expression levels were normalized to HPRT1. Error bars represent S.D. B , dexamethasone( Dex , 10 -3 M)treatmentfor6hofhumanskinleadstoa4-foldinduction of 11 𝛃 -HSD2 ( n = 3). Quantitative real time PCR for the expression of 11 𝛃 -HSD2is shown. Expression levels were normalized to HPRT1. Error bars represent S.D.CC-BY · From the PDF
FIGURE7 Inhibitionofcortisolproductionduringwoundhealingleadsto induction of IL-1 𝛃 expression. Acute wound explants were treated with metyrapone (10 -3 M), dexamethasone ( DEX , 10 -3 M), or combination for 48 h ( n = 3). Metyrapone treatment induces expression of IL-1 𝛃 , whereas dexamethasone reverses that effect. Error bars represent S.D.
FIGURE7 Inhibitionofcortisolproductionduringwoundhealingleadsto induction of IL-1 𝛃 expression. Acute wound explants were treated with metyrapone (10 -3 M), dexamethasone ( DEX , 10 -3 M), or combination for 48 h ( n = 3). Metyrapone treatment induces expression of IL-1 𝛃 , whereas dexamethasone reverses that effect. Error bars represent S.D.CC-BY · From the PDF
Image 17
CC-BY · From the PDF

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 6

  1. Pastar, 2008, Surg. Technol. Int., 17, 105
  2. Steinhoff, 2001, Curr. Opin. Allergy Clin. Immunol., 1, 469
  3. Zouboulis, 2000, Horm. Res., 54, 230
  4. Davis, 2008, Ostomy Wound Manage., 54, 16
  5. Kayes-Wandover, 2000, J. Clin. Endocrinol. Metab., 85, 2519
  6. Sapolsky, 2000, Endocr. Rev., 21, 55

Cited by

Sorted by citations

See all 197

Related

See all

Metadata

Authors
Vukelic, S., Stojadinovic, O., Pastar, I., Rabach, M., Krzyzanowska, A., Lebrun, E., Davis, S. C., Resnik, S., Brem, H., & Tomic-Canic, M.
Affiliations
  • Lenox Hill Hospital (US)
  • University of Miami (US)
  • SUNY Downstate Health Sciences University (US)
  • Hospital for Special Surgery (US)
  • NYU Langone Health (US)
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2011-03
Volume
286
Issue
12
Pages
10265-10275
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.m110.188268
PMC
PMC3060481
OpenAlex
W2080660266
Cited by
193
References
67
Citations by year
2026: 6 · 2025: 12 · 2024: 7 · 2023: 6 · 2022: 12 · 2021: 11 · 2020: 18 · 2019: 9 · 2018: 11 · 2017: 18 · 2016: 17 · 2015: 8 · 2014: 15 · 2013: 19 · 2012: 13
FWCI
18.81
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health
  • National Institute of Nursing Research (R01NR008029, NR008029)
  • National Institute of Diabetes and Digestive and Kidney Diseases (K24DK090135)
Topics
  • Health Sciences › Medicine › Surgery › Wound Healing and Treatments
  • Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
  • Health Sciences › Medicine › Pharmacology (medical) › Inflammatory mediators and NSAID effects
Keywords
cortisol production, epidermis, keratinocytes, wound healing, IL-1β, tissue damage, CYP11B1, 11β-hydroxysteroid dehydrogenase type 2, glucocorticoid receptor, ACTH, metyrapone, negative feedback regulation, inflammatory cytokines
MeSH terms
Adrenocorticotropic Hormone, Animals, Enzyme Inhibitors, Epidermis, Hormones, Humans, Hydrocortisone, Inflammation, Metyrapone, Steroid 11-beta-Hydroxylase, Swine, Wound Healing, Keratinocytes, Interleukin-1beta

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review