Yonsei Med J.  2017 Nov;58(6):1204-1210. 10.3349/ymj.2017.58.6.1204.

11β-Hydroxysteroid Dehydrogenase Type 1 Inhibition Attenuates the Adverse Effects of Glucocorticoids on Dermal Papilla Cells

• Lee SE
• Lee EY
• Kang SJ
• Lee SH

• KMID: 2418903
• DOI: http://doi.org/10.3349/ymj.2017.58.6.1204

Abstract

PURPOSE
Glucocorticoids, stress-related hormones, inhibit hair growth. Intracellular glucocorticoid availability is regulated by 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1). 11β-HSD1 was recently detected in keratinocytes and fibroblasts. However, the expression of 11β-HSD1 in human hair follicles remains unknown. We aimed to examine 11β-HSD1 expression in human dermal papilla cells (DPCs) and to investigate whether modulation of 11β-HSD1 activity can regulate the negative effects of glucocorticoids on DPCs.
MATERIALS AND METHODS
11β-HSD1 expression in normal human scalp skin was examined by immunohistochemistry. 11β-HSD1 protein was detected in Western blots of human DPCs. Cultured human DPCs were treated with cortisol with or without a selective 11β-HSD1 inhibitor and subsequently stained for Ki-67 antibody. Expression levels of 11β-HSD1, Wnt5a, alkaline phosphatase (ALP), and vascular endothelial growth factor (VEGF) were analyzed by Western blotting.
RESULTS
11β-HSD1 was detected in dermal papilla in human scalp skin by immunohistochemistry. Human DPCs expressed 11β-HSD1 protein in vitro. Furthermore, cortisol stimulated the expression of 11β-HSD1 in DPCs. Glucocorticoids decreased cellular proliferation and the expression of Wnt5a, ALP, and VEGF in DPCs. A specific 11β-HSD1 inhibitor significantly attenuated the anti-proliferative effects of cortisol and reversed the cortisol-induced suppression of Wnt5a, ALP, and VEGF expression in DPCs.
CONCLUSION
Our data demonstrated the expression of 11β-HSD1 in human DPCs and revealed that inhibition of 11β-HSD1 activity can partially prevent the negative effect of glucocorticoids on DPCs, suggesting the possible application of 11β-HSD1 inhibitors for stress-related hair loss.

Keyword

11β-hydroxysteroid dehydrogenase type 1; glucocorticoids; dermal papilla cells

MeSH Terms

11-beta-Hydroxysteroid Dehydrogenase Type 1/*antagonists & inhibitors/genetics
Blotting, Western
Cell Proliferation/*drug effects
Cells, Cultured
Fibroblasts/*metabolism
Glucocorticoids/*pharmacology
Humans
Hydrocortisone
Keratinocytes/*metabolism
Signal Transduction
Vascular Endothelial Growth Factor A
Glucocorticoids
Vascular Endothelial Growth Factor A
11-beta-Hydroxysteroid Dehydrogenase Type 1
Hydrocortisone

Figure

• 

  Fig. 1 Expression of 11β-HSD1 in human scalp hair follicles by immunohistochemistry. 11β-HSD1 immunoreactivity was found in ORS keratinocytes (A), MK, and the DP in human scalp hair follicles (B-E) in situ. 11β-HSD1, 11β-hydroxysteroid dehydrogenase type 1; ORS, outer root sheath; MK, matrix keratinocytes; DP, dermal papilla. Scale bar=50 µm.

• 

  Fig. 2 Western blot analysis of 11β-HSD1 expression in unstimulated and cortisol-stimulated human DPCs. Bars show the results of densitometric analysis of the 11β-HSD1 protein band relative to the corresponding GAPDH protein band. Results are presented as mean±SD. *p<0.05. 11β-HSD1, 11β-hydroxysteroid dehydrogenase type 1; DPC, dermal papilla cells; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

• 

  Fig. 3 The effect of a selective 11β-HSD1 inhibitor on the proliferation of cortisol-stimulated human DPCs. (A) Human DPCs were treated with 10-7 M cortisol for 48 hours with or without 30 min pretreatment with 385581 (100 nmol L-1), and immunofluorescent staining was performed with anti-Ki-67 antibody. Nuclei were counterstained with DAPI (Scale bar, 200 µm). (B) The percentage of Ki-67-positive human DPCs (green fluorescence in nuclei) under at least five high power fields in each slide was counted, and statistical analysis was performed using a pair test. Results are presented as mean±SD. *p<0.05, †p<0.01. 11β-HSD1, 11β-hydroxysteroid dehydrogenase type 1; DPC, dermal papilla cells; DAPI, 4′,6′-diamidino-2-phenylindole.

• 

  Fig. 4 The effect of a selective 11β-HSD1 inhibitor on the expression of Wnt5a, ALP, and VEGF in cortisol-stimulated human DPCs. Human DPCs were pre-treated with 100 nmol L-1 11β-HSD1 inhibitor for 30 minutes and stimulated with 10-7 M cortisol for 48 hours. (A) After stimulation, the expression of Wnt5a, ALP, and VEGF was analyzed by Western blot. (B) Bars show the results of densitometric analysis relative to the corresponding GAPDH protein band. Results are presented as mean±SD. *p<0.05, †p<0.01. 11β-HSD1, 11β-hydroxysteroid dehydrogenase type 1; ALP, alkaline phosphatase; VEGF, vascular endothelial growth factor; DPC, dermal papilla cells; GAPDH, glyceraldehyde 3-phosphate dehydrogenase.

Reference

1. Alexopoulos A, Chrousos GP. Stress-related skin disorders. Rev Endocr Metab Disord. 2016; 17:295–304.

2. Taheri R, Behnam B, Tousi JA, Azizzade M, Sheikhvatan MR. Triggering role of stressful life events in patients with alopecia areata. Acta Dermatovenerol Croat. 2012; 20:246–250.

3. Botchkarev VA. Stress and the hair follicle: exploring the connections. Am J Pathol. 2003; 162:709–712.

4. Peters EM, Arck PC, Paus R. Hair growth inhibition by psychoemotional stress: a mouse model for neural mechanisms in hair growth control. Exp Dermatol. 2006; 15:1–13.

5. Ito N, Ito T, Kromminga A, Bettermann A, Takigawa M, Kees F, et al. Human hair follicles display a functional equivalent of the hypothalamic-pituitary-adrenal axis and synthesize cortisol. FASEB J. 2005; 19:1332–1334.

6. Zhang X, Yu M, Yu W, Weinberg J, Shapiro J, McElwee KJ. Development of alopecia areata is associated with higher central and peripheral hypothalamic-pituitary-adrenal tone in the skin graft induced C3H/HeJ mouse model. J Invest Dermatol. 2009; 129:1527–1538.

7. Novak MA, Hamel AF, Coleman K, Lutz CK, Worlein J, Menard M, et al. Hair loss and hypothalamic-pituitary-adrenocortical axis activity in captive rhesus macaques (Macaca mulatta). J Am Assoc Lab Anim Sci. 2014; 53:261–266.

8. Choi EH, Demerjian M, Crumrine D, Brown BE, Mauro T, Elias PM, et al. Glucocorticoid blockade reverses psychological stress-induced abnormalities in epidermal structure and function. Am J Physiol Regul Integr Comp Physiol. 2006; 291:R1657–R1662.

9. Demerjian M, Choi EH, Man MQ, Chang S, Elias PM, Feingold KR. Activators of PPARs and LXR decrease the adverse effects of exogenous glucocorticoids on the epidermis. Exp Dermatol. 2009; 18:643–649.

10. Paus R, Handjiski B, Czarnetzki BM, Eichmüller S. A murine model for inducing and manipulating hair follicle regression (catagen): effects of dexamethasone and cyclosporin A. J Invest Dermatol. 1994; 103:143–147.

11. Pérez P, Page A, Bravo A, Del Río M, Giménez-Conti I, Budunova I, et al. Altered skin development and impaired proliferative and inflammatory responses in transgenic mice overexpressing the glucocorticoid receptor. FASEB J. 2001; 15:2030–2032.

12. Choi SJ, Cho AR, Jo SJ, Hwang ST, Kim KH, Kwon OS. Effects of glucocorticoid on human dermal papilla cells in vitro. J Steroid Biochem Mol Biol. 2013; 135:24–29.

13. Tomlinson JW, Walker EA, Bujalska IJ, Draper N, Lavery GG, Cooper MS, et al. 11beta-hydroxysteroid dehydrogenase type 1: a tissue-specific regulator of glucocorticoid response. Endocr Rev. 2004; 25:831–866.

14. Tiganescu A, Walker EA, Hardy RS, Mayes AE, Stewart PM. Localization, age- and site-dependent expression, and regulation of 11β-hydroxysteroid dehydrogenase type 1 in skin. J Invest Dermatol. 2011; 131:30–36.

15. Lee SE, Kim JM, Jeong MK, Zouboulis CC, Lee SH. 11β-hydroxysteroid dehydrogenase type 1 is expressed in human sebaceous glands and regulates glucocorticoid-induced lipid synthesis and toll-like receptor 2 expression in SZ95 sebocytes. Br J Dermatol. 2013; 168:47–55.

16. Terao M, Katayama I. Local cortisol/corticosterone activation in skin physiology and pathology. J Dermatol Sci. 2016; 84:11–16.

17. Yang CC, Cotsarelis G. Review of hair follicle dermal cells. J Dermatol Sci. 2010; 57:2–11.

18. Katsuoka K, Schell H, Wessel B, Hornstein OP. Effects of epidermal growth factor, fibroblast growth factor, minoxidil and hydrocortisone on growth kinetics in human hair bulb papilla cells and root sheath fibroblasts cultured in vitro. Arch Dermatol Res. 1987; 279:247–250.

19. Hembree JR, Harmon CS, Nevins TD, Eckert RL. Regulation of human dermal papilla cell production of insulin-like growth factor binding protein-3 by retinoic acid, glucocorticoids, and insulin-like growth factor-1. J Cell Physiol. 1996; 167:556–561.

20. Reddy S, Andl T, Bagasra A, Lu MM, Epstein DJ, Morrisey EE, et al. Characterization of Wnt gene expression in developing and postnatal hair follicles and identification of Wnt5a as a target of Sonic hedgehog in hair follicle morphogenesis. Mech Dev. 2001; 107:69–82.

21. Rendl M, Lewis L, Fuchs E. Molecular dissection of mesenchymal-epithelial interactions in the hair follicle. PLoS Biol. 2005; 09. 20. DOI: 10.1371/journal.pbio.0030331. [Epub].

22. Iida M, Ihara S, Matsuzaki T. Hair cycle-dependent changes of alkaline phosphatase activity in the mesenchyme and epithelium in mouse vibrissal follicles. Dev Growth Differ. 2007; 49:185–195.

23. McElwee KJ, Kissling S, Wenzel E, Huth A, Hoffmann R. Cultured peribulbar dermal sheath cells can induce hair follicle development and contribute to the dermal sheath and dermal papilla. J Invest Dermatol. 2003; 121:1267–1275.

24. Ohyama M, Kobayashi T, Sasaki T, Shimizu A, Amagai M. Restoration of the intrinsic properties of human dermal papilla in vitro. J Cell Sci. 2012; 125(Pt 17):4114–4125.

25. Feig PU, Shah S, Hermanowski-Vosatka A, Plotkin D, Springer MS, Donahue S, et al. Effects of an 11β-hydroxysteroid dehydrogenase type 1 inhibitor, MK-0916, in patients with type 2 diabetes mellitus and metabolic syndrome. Diabetes Obes Metab. 2011; 13:498–504.

26. Morgan SA, McCabe EL, Gathercole LL, Hassan-Smith ZK, Larner DP, Bujalska IJ, et al. 11β-HSD1 is the major regulator of the tissue-specific effects of circulating glucocorticoid excess. Proc Natl Acad Sci U S A. 2014; 111:E2482–E2491.

27. Youm JK, Park K, Uchida Y, Chan A, Mauro TM, Holleran WM, et al. Local blockade of glucocorticoid activation reverses stress- and glucocorticoid-induced delays in cutaneous wound healing. Wound Repair Regen. 2013; 21:715–722.