Krebiehl, G.

Mihály, Johanna

Rühl, Ralph

Sonntag, D.

Szegedi, Andrea

Töröcsik, Dániel

British Journal of Dermatology

University of Debrecen Electronic Archive

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British Association of Dermatologists 
This article is protected by copyright. All rights reserved. 
 
Received Date : 24-Mar-2014 
Revised Date   : 17-Jun-2014 
Accepted Date : 18-Jun-2014 
Article type      : Correspondence 
 
 
Steroid concentrations in atopic dermatitis patients: 
Reduced plasma DHEAS and increased cortisone levels 
 
 
J. Mihály1, D. Sonntag2, G. Krebiehl2, A. Szegedi3, 
D. Töröcsik4, R. Rühl1,5  
 
 
1 Department of Biochemistry and Molecular Biology, University of Debrecen;  
2 BIOCRATES Life Sciences AG, Innsbruck, Austria; 
3 Department of Dermatological Allergology, University of Debrecen;  
4 Department of Dermatology, University of Debrecen; 
5 Paprika Bioanalytics BT, Debrecen, Hungary. 
 
 
Corresponding author: 
 
Dr. Ralph Rühl 
Department of Biochemistry and Molecular Biology 
Medical and Health Science Center 
University of Debrecen 
Nagyerdei krt. 98 
H-4032 Debrecen 
This article is protected by copyright. All rights reserved. 
Tel: +36-30 2330 501 
Fax: +36 52 314 989 
E-mail: ralphruehl@web.de 
 
DEAR EDITOR, atopic dermatitis (AD) is a chronically relapsing inflammatory skin 
disease, which is characterized by a disrupted epidermal barrier function present 
both in affected skin and in non-affected skin (5). Mainly glucocorticosteroids were 
used in topical and systemic atopy treatments because of their potent anti-
inflammatory effects (8), unfortunately with strong side effects (1). Other steroid 
hormones have also been reported to have supportive or detrimental effects on 
atopy (2, 12).  
 
In this study we aimed to investigate how the levels of several steroids are altered in 
the plasma of AD-patients compared to healthy volunteers. 
 
Plasma samples of 20 healthy volunteers and 20 AD-patients were collected at 
comparable day time, between 8-10 a.m. in order to minimise the influence of 
circadian regulation of hormones. In addition, AD-patients were not under treatment 
of oral glucocorticoids and topical corticosteroids for at least 5 days prior to blood 
sampling. SCORAD (SCORe Atopic Dermatitis) index of AD-patients was 35.2 (range 
13-64) (11). The sterroid hormones were quantified with the commercially available 
AbsoluteIDQ® Stero17 kit from Biocrates that comprises 16 endogenous steroids 
(Table 1). 200-450 µL of human plasma were used for analysis. The HPLC-MS/MS 
analysis in multiple reaction monitoring mode (MRM) using a SCIEX API 4000 QTrap 
was preceeded by a sample preparation procedure based on solid phase extraction 
technique in a 96 well plate-format fundamental for pre-cleaning and pre-
concentration of the target steroid hormones. For quantification of the steroid 
compounds, 7-point external calibration curves and 13 isotope-labeled internal 
standards were used.  
 
Steroid hormones can be divided into five different categories; a) 
mineralocorticosteroids, b) glucocorticosteroids, c) androgens, d) estrogens and e) 
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progestagens. Mineralocorticoid, progestagen levels remained mainly unchanged in 
AD-patients vs healthy volunteers (table 1). From the glucocorticoids the level of 
cortisone was significantly increased (H: 66.2 ± 17.5 ng/ml, D: 79.7 ± 21.5 ng/ml, 
p=0.04) while the bioactive metabolite cortisol also displayed a slight tendency of 
increase (H: 327 ± 154 ng/ml, D: 407 ± 209 ng/ml, p=0.19). The plasma level of 
the androgen DHEAS was significantly decreased (H: 8857 ± 3918 ng/ml to D: 5187 
± 3362 ng/ml) in AD-patients. Regarding the levels of the estrogen estrone a 
tendency of decreased levels could be found in AD-patients (H: 0.33 ± 0.14 ng/ml, 
D: 0.25 ± 0.11 ng/ml, p=0.07). When comparing steroid concentrations in healthy 
male (HM) volunteers and male AD-patients (DM) (table 2A), the mineralocorticoid, 
estrogen and progestagen levels were comparable in HM vs DM, while for the 
androgen DHEAS levels (HM: 10141 ± 4758 ng/ml, DM: 5186 ± 3612 ng/ml) were 
significantly lower and DHEA levels just displayed a lower tendency (HM: 22.6 ± 7.4 
ng/ml, DM: 15.4 ± 7.5 ng/ml, p=0.08) in the plasma of DM. Levels of the steroids 
mineralocorticoid, estrogens and progestagen in healthy female volunteers (HF) and 
female AD-patients (DF) (table 2B) were comparable between HF and DF, while the 
androgen DHEAS levels (HW: 8166 ± 3305 ng/ml, DF: 5188 ± 3323 ng/ml) were 
significantly lower in plasma of female AD-patients. The glucocorticoid levels of 11-
deoxycortisol (HW: 0.47 ± 0.26 ng/ml, DF: 1.16 ± 0.81 ng/ml) were significantly 
higher and cortisone levels displayed just a higher tendency (HF: 66.0 ± 17.3 ng/ml, 
DF: 79.5 ± 17.4 ng/ml, p=0.07) in female AD-patients. Steroid concentration of DHT 
(M: 0.82 ± 0.41 ng/ml, F: 0.56 ± 0.29 ng/ml) and testosterone (M: 11.1 ± 8.39 
ng/ml, F: 3.15 ± 4.94 ng/ml) were significantly lower in female individuals (F) vs 
male individuals (M) (table 2C), while ethiocolanolone levels (M: 0.37 ± 0.17 ng/ml, 
F: 0.58 ± 0.39 ng/ml) were significantly lower in male individuals. Mineralocorticoid, 
glucocorticoid, estrogen and progestagen levels were surprisingly comparable in 
woman and man. 
 
In summary in this study we determined that 2 out of 16 steroids were significantly 
different in healthy volunteers vs AD-patients. Cortisone, which is higher in AD-
patients plasma, is a direct precursor of the bioactive corticosteroid cortisol, which 
just displays a higher tendency and is known for its potent anti-inflammatory effects 
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(8). These increased levels of corticosteroids levels in AD-patients plasma may mean 
that there is an increased formation of anti-inflammatory steroids as a feedback 
mechanism present in AD-patients which comparably has also been found for 
precursors of anti-inflammatory eicosanoids (11). In addition a tendency of reduced 
levels of the anti-inflammatory ERα ligand estrone (6) was found in AD-patients. 
DHEA is a precursor of testosterone, its levels just display a lower tendency in male 
AD-patients, while its sulfonation metabolite DHEAS is lower in male and female AD-
patients and decreased DHEAS levels were previously found in atopic allergy and 
chronic urticaria (8-9). In acne patients increased levels of DHEAS were found (3) 
and further impaired sebum secretion may be the outcome of this decreased DHEAS 
levels and may even could contribute to AD-phenotype (4). Furthermore DHEA(S) 
application affects production and secretion of Th1 and Th2 cytokines showing an 
immunomodulatory effect during allergic sensitization and allergic responses (9-10). 
The DHEA sulfonation is mediated by dehydroepiandrosterone sulfotransferase 
(SULT2A1), which is a direct vitamin D receptor target gene (7) and thereby 
suggests reduced vitamin D-signaling during atopy (13). 
 
We conclude that altered steroid levels in the plasma of AD-patients indicate altered 
vitamin D signaling (based on reduced DHEA sulfonation) and increased feedback for 
anti-inflammatory signaling (increased levels of cortisone) present in AD-patients.  
 
Funding sources: RR and DT are members of the COST projects “Mast Cells and 
Basophils - Targets for innovative therapies”. The work was also supported by 
TÁMOP-4.2.2.A-11/1/KONV-2012-0023 "VÉD-ELEM" project. The project is 
implemented through the New Hungary Development Plan co-financed by the 
European Social Fund and the European Regional Development Fund. In addition 
these projects were funded by the OTKA projects (AS: OTKA K 108421 and RR: 
OTKA K 109362). 
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Table 1. Concentration of mineralocorticoids, glucocorticoids, androgens, estrogens and 
progestagen in healthy volunteers vs AD-patients. SD - standard deviation. Data are shown as 
mean and standard deviation based on n=20 samples. Using a student ttest a p value of less than 
0.05 was considered significant. 
 
Steroid concentrations in healthy volunteers (H, n=20) and AD-patients (D, n=20) in ng/ml. 
 
Healthy volunteers AD-patients   
mean ± SD  mean ± SD   Significance 
Mineralocorticoids 
11-deoxycorticosterone  0.76 ± 0.39  65.5 ± 200  0.17 
aldosterone   0.51 ± 0.39  0.27 ± 0.41  0.06 
Glucocorticoids 
11-deoxycortisol   0.79 ± 0.78  0.98 ± 0.65  0.45 
cortisone    66.2 ± 17.5  79.7 ± 21.5  0.04  
corticosterone   13.4 ± 16.0  10.1 ± 8.09  0.45 
cortisol    327 ± 154  407 ± 209  0.19 
Androgens 
androstenedione   4.74 ± 1.89  4.28 ± 1.73  0.44 
androsterone   1.40 ± 0.70  1.11 ± 0.48  0.13 
DHEA    21.8 ± 9.38  21.2 ± 14.4  0.95 
DHEAS    8857 ± 3918  5187 ± 3362  <0.01 
DHT    0.71 ± 0.35  0.61 ± 0.38  0.34 
etiocholanolone   0.54 ± 0.43  0.45 ± 0.22  0.40 
testosterone   6.63 ± 8.31  6.04 ± 7.05  0.72 
Estrogens 
E1 (estrone)   0.33 ± 0.14  0.25 ± 0.11  0.07 
E2 (estradiol)   0.58 ± 0.46  0.43 ± 0.21  0.22 
Progestagen 
progesterone   2.02 ± 6.04  2.53 ± 9.14  0.87 
 
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Table 2: Concentration of mineralocorticoids, glucocorticoids, androgens, estrogens and 
progestagen in (A.) healthy male volunteers (HM) vs male AD-patients (DM), (B.) healthy female 
volunteers (HF) vs female AD-patients (DF) as well as (C.) female individuals (F, n=24) vs male 
individuals (M, n=16). SD - standard deviation. 
 
A. Steroid concentrations in healthy male volunteers (HM, n=7) and male AD-patients (DM, n=9) in 
ng/ml.  
 
Healthy male  Male AD-patients  
mean ± SD  mean ± SD   Significance 
Mineralocorticoids 
11-deoxycorticosterone  0.93 ± 0.31  124 ± 290  0.28 
aldosterone   0.36 ± 0.31  0.21 ± 0.17  0.24 
Glucocorticoids 
11-deoxycortisol   1.38 ± 1.03  0.75 ± 0.27  0.10 
cortisone    66.6 ± 19.2  79.9 ± 26.7  0.29  
corticosterone   16.4 ± 18.4  8.72 ± 7.00  0.27 
cortisol    332 ± 168  389 ± 264  0.63 
Androgens  
androstenedione   4.17 ± 1.34  3.86 ± 1.08  0.61 
androsterone   1.39 ± 0.69  1.07 ± 0.55  0.32 
DHEA    22.6 ± 7.44  15.4 ± 7.53  0.08 
DHEAS    10141 ± 4758  5186 ± 3612  0.03 
DHT    0.90 ± 0.25  0.76 ± 0.50  0.52 
etiocholanolone   0.33 ± 0.15  0.40 ± 0.19  0.41 
testosterone   14.1 ± 7.50  8.81 ± 8.73  0.23 
Estrogens 
E1 (estrone)   0.29 ± 0.12  0.25 ± 0.07  0.42 
E2 (estradiol)   0.45 ± 0.19  0.50 ± 0.29  0.70 
Progestagen 
progesterone   0.35 ± 0.19  0.58 ± 0.97  0.56 
 
B. Steroid concentrations in healthy female volunteers (HF, n=13) and female AD-patients (DF, 
n=11) in ng/ml.  
 
Healthy female  Female AD-patients Significance 
Mineralocorticoids 
11-deoxycorticosterone  0.67 ± 0.41  17.7 ± 56.5  0.29 
aldosterone   0.59 ± 0.40  0.32 ± 0.54  0.18 
Glucocorticoids  
11-deoxycortisol   0.47 ± 0.26  1.16 ± 0.81  0.01 
cortisone    66.0 ± 17.3  79.5 ± 17.4  0.07  
corticosterone   11.8 ± 14.5  11.1 ± 9.08  0.89 
cortisol    323 ± 147  421 ± 164  0.14 
Androgens 
androstenedione   5.05 ± 2.04  4.62 ± 2.11  0.62 
androsterone   1.40 ± 0.70  1.15 ± 0.43  0.31 
DHEA    21.3 ± 11.3  25.9 ± 17.2  0.45 
DHEAS    8166 ± 3305  5188 ± 3323  0.04 
DHT    0.61 ± 0.35  0.49 ± 0.20  0.31 
etiocholanolone   0.65 ± 0.48  0.49 ± 0.25  0.34 
testosterone   2.63 ± 5.36  3.76 ± 4.56  0.59 
Estrogens 
E1 (estrone)   0.35 ± 0.15  0.26 ± 0.14  0.12 
E2 (estradiol)   0.65 ± 0.53  0.38 ± 0.11  0.11 
Progestagen 
progesterone   2.92 ± 7.24  4.14 ± 12.3  0.77 
 
C. Steroid concentration distribution in female individuals (F, n=24) vs male individuals (M, n=16) 
in ng/ml. 
 
Male individuals  Female individuals Significance 
Mineralocorticoids 
11-deoxycorticosterone  70.2 ± 221  8.45 ± 38.2  0.19 
aldosterone   0.28 ± 0.24  0.47 ± 0.48  0.15 
Glucocorticoids 
11-deoxycortisol   1.03 ± 0.76  0.79 ± 0.67  0.30 
cortisone    74 ± 24  72.2 ± 18.3  0.78  
corticosterone   12.1 ± 13.3  11.5 ± 12.0  0.89 
cortisol    364 ± 222  368 ± 159  0.94 
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Androgens 
androstenedione   3.99 ± 1.17  4.85 ± 2.04  0.14 
androsterone   1.21 ± 0.62  1.29 ± 0.59  0.70 
DHEA    18.6 ± 8.11  23.4 ± 14.2  0.22 
DHEAS    7354 ± 4739  6801 ± 3578  0.68 
DHT    0.82 ± 0.41  0.56 ± 0.29  0.02 
etiocholanolone   0.37 ± 0.17  0.58 ± 0.39  <0.05 
testosterone   11.1 ± 8.39  3.15 ± 4.94  <0.01 
Estrogens 
E1 (estrone)   0.27 ± 0.09  0.31 ± 0.15  0.36 
E2 (estradiol)   0.48 ± 0.25  0.53 ± 0.41  0.67 
Progestagen 
progesterone   0.48 ± 0.73  3.48 ± 9.68  0.23 
 
 
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Steroid concentrations in atopic dermatitis patients: reduced plasma DHEAS and increased cortisone levels

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Steroid concentrations in atopic dermatitis patients: reduced plasma DHEAS and increased cortisone levels

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