Dihydrotestosterone-Inducible IL-6 Inhibits Elongation of Human Hair Shafts by Suppressing Matrix Cell Proliferation and Promotes Regression of Hair Follicles.

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Dihydrotestosterone-Inducible IL-6 Inhibits Elongation of Human Hair Shafts by Suppressing Matrix Cell Proliferation and Promotes Regression of Hair Follicles in Mice Mi Hee Kwack, Ji Sup Ahn, Moon Kyu Kim, Jung Chul Kim, Young Kwan Sung Journal of Investigative Dermatology Volume 132, Issue 1, Pages 43-49 (January 2012) DOI: 10.1038/jid.2011.274 Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Upregulation of IL-6 in cultured balding DP cells compared with the non-balding DP cells and IL-6 induction in response to DHT in balding DP cells. Seven pairs of matched balding and non-balding dermal papilla (DP) cells were analyzed by reverse transcription (RT)-PCR (a). IL-6 levels in conditioned medium of balding (B) and non-balding (NB) DP cells from four matched pairs in duplicate measurements were made by ELISA and the relative fold change of IL-6 in each cases (B/NB) are shown (b). Balding DP cells were treated with varying concentrations of dihydrotestosterone (DHT), and concentrations of IL-6 were measured by ELISA (c); values represent the mean±SD of five experiments using five different DP cell strains and duplicate measurements were made for each experiment (*P<0.005, **P<0.001). Cells were also treated with varying concentrations of DHT for 6hours (d) or 100nM DHT for varying times (e) and analyzed by RT-PCR. Journal of Investigative Dermatology 2012 132, 43-49DOI: (10.1038/jid.2011.274) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Expression of IL-6R and gp130 in hair follicles and cultured cells. Human scalp hair follicles were immunostained with antibodies against IL-6 receptor (IL-6R, a) and glycoprotein 130 (gp130, b). High-powered images of IL-6R and gp130 expression in hair bulb regions are also shown in c and d, respectively. Close-up images of boxed regions of c and d with emphasis on the follicular matrix are also shown in e and f. Bar=0.1mm. Immunoblot analysis (g) of IL-6R and gp130 protein expression in cultured hair cells. Actin expression was measured to check the quantity and integrity of the protein samples. DP, dermal papilla; DS, dermal sheath; ORS, outer root sheath. Journal of Investigative Dermatology 2012 132, 43-49DOI: (10.1038/jid.2011.274) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Immunohistochemical staining for IL-6R and gp130 in balding versus non-balding hair follicles. Matched balding (a and e) and non-balding (b and f) scalp hair follicles were immunostained with antibodies against IL-6R (a and b) and gp130 (e and f). Close-up images of boxed regions of a, b, e, and f with emphasis on the follicular matrix are also shown in c, d, g, and h. gp130, glycoprotein 130; IL-6R, IL-6 receptor. Journal of Investigative Dermatology 2012 132, 43-49DOI: (10.1038/jid.2011.274) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Effect of rhIL-6 on cultured human hair follicles. Isolated human hair follicles were cultured for 6 days in the absence or presence of 5 or 50ngml−1 recombinant human IL-6 (rhIL-6), and hair shaft elongation was measured (a). Values are average of four mean±SD of six determinations per experiment from four experiments (*P<0.05). Human hair follicles were treated in the absence (c and e) or presence of 50ngml−1 rhIL-6 (d and f) for 2 days and stained with Ki-67 immunofluorescence staining. Corresponding 4,6-diamidino-2-phenylindole (DAPI) nuclear staining is also shown (e and f). Ki-67-positive proliferative cells (red) in the hair bulb were counted and data are mean±SD from five hair follicles (*P<0.05; b). Bar=0.1mm. Journal of Investigative Dermatology 2012 132, 43-49DOI: (10.1038/jid.2011.274) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Effect of rhIL-6 on catagen onset. Scheme of the experiment (a). The back skins of C57BL/6 mice (n=6) were shaved at 5 weeks of age and the vehicle (b) or 100μl of 500ngml−1 of recombinant human IL-6 (rhIL-6; c) was injected once a day for 3 days. The skin samples were stained with hematoxylin and eosin and a representative histology is shown. The same experiment was repeated twice using different mice. Bar=0.1mm. Journal of Investigative Dermatology 2012 132, 43-49DOI: (10.1038/jid.2011.274) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Proposed model based on this study. Dihydrotestosterone (DHT)-inducible IL-6 from dermal papilla functions as an inhibitory paracrine mediator that inhibits hair shaft growth by suppressing matrix cell proliferation. Journal of Investigative Dermatology 2012 132, 43-49DOI: (10.1038/jid.2011.274) Copyright © 2012 The Society for Investigative Dermatology, Inc Terms and Conditions