DOI:10.1111/j.1600-0625.2012.01495.

x
www.blackwellpublishing.com/EXD
Original Article

Influence of chemical peeling on the skin stress response system

Ayako Kimura1, Nobuo Kanazawa2, Hong-Jin Li2, Nozomi Yonei1, Yuki Yamamoto2 and Fukumi Furukawa2
1
Department of Dermatology, Naga Municipal Hospital, Wakayama, Japan; 2Department of Dermatology, School of Medicine, Wakayama Medical
University, Wakayama, Japan
Correspondence: Ayako Kimura, MD, Department of Dermatology, Naga Municipal Hospital, 1282 Uchita, Kinokawa, Wakayama 649-6414, Japan,
Tel.: +81-736-77-2019, Fax: +81-73677-4659, e-mail: kimura@nagahp.jp

Abstract: Skin stress response system (SSRS) involves
corticotropin-releasing hormone (CRH) and proopiomelanocortin
(POMC)-derived peptides, such as adrenocorticotropic hormone
(ACTH), a-melanocyte-stimulating hormone (MSH) and b-
endorphin that are locally generated in response to locally
provided stressors or proinflammatory cytokines. This system
would restrict tissue damage and restore local homoeostasis.
Trichloroacetic acid (TCA) is one of the most widely used peeling
agents and applied for cosmetic treatment of photodamaged skin.
However, the biological mechanism responsible for TCA peeling
has yet to be fully determined. While our investigation focused on
the inflammation and wound healing pathways, in the recent
study, we have examined involvement of the SSRS as the third
pathway. Mostly depending on our findings that TCA peeling
activates the SSRS by inducing the POMC expression of
keratinocytes in the CRH-independent manner, together with the
results reported by other researchers, we can say that the
biological effect of POMC seems to be responsible for the TCA-
induced epidermal SSRS activation.
Key words: chemical peeling – proopiomelanocortin – skin stress
response system – trichloroacetic acid
Accepted for publication 20 March 2012

Introduction These hormones induce a variety of reactions to stress. For

Chemical peeling is a rejuvenation method in which chemical example, CRH is able to activate the sympathetic system, restrain
regents are applied on the skin surface, thereby improving the the parasympathetic system, cause awakening and uneasiness and
appearance of photoaged skin by reducing features such as actinic decrease appetite. MSH stimulates the production of melanin,
lentigines and wrinkle. Peeling regents include acids and phenol inhibits fever and inflammation and controls fat accumulation.
and have a protein coagulation effect, thus causing tissue injury to Endorphin participates in sedation, blood pressure regulation and
various depths of skin. Trichloroacetic acid (TCA) is one of the most temperature control (3). ACTH further induces production and
widely used peeling agents and induces full necrosis of the whole epi- secretion of cortisol from the adrenal cortex through activation of
dermis, followed by reconstitution of the epidermis and the matrix melanocortin receptors (MCRs). Cortisol has a regulatory func-
of the papillary dermis (1). However, the biological mechanism tion, terminating the release of upstream hormones (CRH and
responsible for TCA peeling has yet to be fully determined. POMC) through feedback mechanisms. In cases of inflammation,
We previously investigated molecular mechanisms underlying this pathway can be activated by proinflammatory cytokines to
chemical peeling, using TCA peeling as a model, focusing on the
inflammation and wound healing pathways. In the recent study,
we have examined involvement of the skin stress response system Systemic stress
(SSRS), a skin-specific stress response endocrine system, as the
third pathway. We performed experiments on cultured keratino-
cytes, as well as mouse and human skin, and have found that
mRNA and protein of proopiomelanocortin (POMC), a hormone Hypothalamus
precursor that is a key molecule in the SSRS, are specifically IL-1
induced in the epidermis by TCA stimulation. In this review, we C H
CRH IL-6
TNFα
briefly summarize the SSRS, in comparison with its systemic Pituitary
counterpart, and such an experimentally revealed influence of
TCA peeling on this system, to discuss significance of the SSRS
POMC
activation in chemical peeling (2).
Hypothalamic–pituitary–adrenal (HPA) axis MSH ACTH β-endorphin
The classical neuroendocrine pathway in response to systemic
stress, known as the hypothalamic–pituitary–adrenal (HPA) axis,
includes the hypothalamic release of corticotropin-releasing hor- Adrenal
mone (CRH), subsequent activation of pituitary CRH receptors Cor sol
(CRHR), and production and release of POMC-derived peptides,
Nega ve feedback
such as adrenocorticotropic hormone (ACTH), a-melanocyte-
stimulating hormone (MSH) and b-endorphin (Fig. 1). Figure 1. Hypothalamic–pituitary–adrenal axis: HPA axis.

ª 2012 John Wiley & Sons A/S

8 Experimental Dermatology, 21 (Suppl. 1), 8–10
 Influence of chemical peeling on the skin stress response system
yield anti-inflammatory effects and to prevent overwhelming
inflammation.
Cytokines such as interleukin (IL)-1, IL-6 and TNF-a, which
are produced by stress-activated immunocytes without mediation
by neural pathways in the brain, also directly stimulate the secre-
tion of CRH in the hypothalamus and POMC-related peptides in
the pituitary. These endocrine glands are thus also affected by the
action of the immune system as a result of inflammation. In this
way, CRH synthesis and secretion in the hypothalamus is pro-
moted by stress stimulation via the central nervous system and
inflammatory cytokines and is inhibited by increased glucocortic-
oids (4,5). Transcription of POMC mRNA in POMC-related pep-
tide secretory cells in the anterior lobe of the pituitary is
promoted by CRH secreted as a result of stress and by inflamma-
tory cytokine stimulation and is inhibited by glucocorticoids. The
body thus possesses important mechanisms for maintaining ho-
moeostasis in the face of psychological and other endogenous
stresses, as well as exogenous stresses such as wounds and infec-
tion, based on the HPA axis.
Skin stress response system
The most common stressors in nature are environmental, as rep-
resented by solar radiation, biological and chemical insults. These
stressors primarily affect the skin, a strategically located protective
barrier. To maintain the integrity of its diverse structures and
functional domains, the skin needs to be able to deal with external
stresses by employing various defense mechanisms. In the skin, an
equivalent system to the HPA axis has been proposed to function
in response to the local stress. This system, known as the SSRS,
involves locally produced CRH and POMC products that are gen-
erated in response to locally provided stressors or proinflammato-
ry cytokines (Fig. 2) (6). This system restricts tissue damage and
restores local homoeostasis. The expression of CRH, CRHR1
and POMC mRNAs has been demonstrated in the pilosebaceous
and sweat gland units of the human skin. In normal human skin,
expression of CRH, CRHR1, melanocortin-1 receptor (MC-1R)
proteins and POMC-related peptides has been demonstrated in
keratinocytes and melanocytes (7). It has been reported that
POMC products are more frequently detected in skin affected by
diseases, such as psoriatic keratinocytes, inflammatory infiltrates in
scarring alopecia, nevocytes, epithelial cell nests of basal cell carci-
noma and melanoma cells (8,9).
It has been reported that ultraviolet B (UVB) and IL-1 stimu-
late production of CRH and POMC, while dexamethasone inhibits
these in human melanocytes and keratinocytes in culture (10,11).
Local stress
Epidermis
IL-1
CRH IL-6
TNFα
POMC
MSH ACTH β-endorphin
Figure 2. Skin Stress Response System: SSRS.
ª 2012 John Wiley & Sons A/S
Experimental Dermatology, 21 (Suppl. 1), 8–10
In a recent report, UVB irradiation directly induces POMC
expression in keratinocytes through p53 activation (12). As
p53-deficient mice do not show the suntan response after UV
irradiation, this pathway is responsible for UV-induced
hyperpigmentation. Interestingly, POMC is also involved in
pathologic hyperpigmentation with p53-inducing chemicals such
as 5-fluoro-uracil (12).
It has been reported that expression of CRH, CRHR1, POMC
and MC-1R proteins fluctuate during synchronized hair follicle
cycling in mice and that their expression is elevated in anagen and
reduced in catagen and telogen. Expression of CRHR1, POMC
and MC-1R mRNA also fluctuates during synchronized hair folli-
cle cycling in mice (13). On the other hand, CRH mRNA is not
detected in all hair follicle cycling. Therefore, it has been suggested
that CRH peptide originates from an extracutaneous site of CRH
synthesis (14). However, in our recent experiments, expression of
CRH mRNA was observed, in addition to CRHR1, POMC and
MC-1R, in mice skin (2). Not all of these proteins expressed in
the skin are solely skin-derived, and it is possible that proteins
produced in the hypothalamus and pituitary may also be distrib-
uted in the skin. The fact that the skin has an ability to transcribe
and translate POMC and MCR-1 genes, however, suggests that
they have some role in the skin-specific responses to local stresses.
The actions of CRH in the skin via the HPA axis and the SSRS
include promotion ⁄ inhibition of cell proliferation and promo-
tion ⁄ inhibition of inflammation; those of the POMC-related pep-
tide MSH include the promotion of melanin production, anti-
inflammatory action, and promotion of keratinocyte proliferation;
and those of ACTH include induction of the growth period in the
hair cycle in addition to the promotion of melanin production in
the same way as MSH (15–18). In addition, b-endorphin is known
to promote the proliferation of keratinocytes and fibroblasts, as
well as enhancing skin turnover and improving its barrier function
(19–21).
Influence of TCA peeling on the SSRS
To determine the influence of TCA peeling on SSRS, we examined
the expressions of POMC, MC-1R, CRH and CRHR1 mRNA by
reverse transcription-polymerase chain reaction in murine plantar
and healthy human abdominal skin specimens after TCA treat-
ment. In addition, expression of their proteins and POMC-derived
peptides were examined immunohistochemically. After TCA treat-
ment, transient upregulation of POMC and MC-1R mRNA
expression was observed in both murine and human skin.
Enhanced POMC protein, recovery of once-impaired MCR-1 pro-
tein, and no enhancement of POMC-derived peptide production
were confirmed immunohistochemically in both murine and
human epidermis. In contrast, expression levels of neither CRH
and CRHR1 mRNA nor epidermal protein were enhanced after
TCA application in either murine or human skin, except for
induction of human CRH mRNA expression. These results suggest
that TCA activates SSRS by inducing POMC and MC-1R produc-
tion of keratinocytes in a CRH-independent manner and that the
biological effects of POMC itself are responsible for TCA-induced
epidermal SSRS activation (2).
POMC is cleaved into ACTH, MSH and b-endorphin in secre-
tion cells by specific enzymes, such as proprotein convertase 1
(PC1) and PC2. On the other hand, POMC is known to be
secreted as whole POMC without cleavage by these enzymes (22).
9
 Kimura et al.

In addition, it has been reported that most of the protein secreted
by keratinocytes and melanocytes is POMC itself, not POMC-
related peptides such as ACTH or MSH. It has been shown that
the biological function of POMC stimulates the production of
melanin through MC-1R, but POMC has a much lower potency
than aMSH (23). After UV irradiation, induction of POMC pro-
tein is only temporal and aMSH is upregulated even after the
reduction of POMC production (12). Thus, although POMC pro-
tein could be involved in the pigmentation process, its contribu-
tion could be minor.
Three pathways in TCA peeling
TCA is one of the most widely used peeling agents and is used for
the cosmetic treatment of photodamaged skin and pitting scars
caused by acne. High concentration of TCA, which can reach the
papillary dermis, induces full necrosis of the whole epidermis, fol-
lowed by reconstitution of the epidermis and the matrix of the
papillary dermis through wound healing processes. Cytotoxic
effects of TCA, such as suppressing proliferation of keratinocytes
and fibroblasts and protein synthesis by fibroblasts, have been
reported previously (24). However, biological mechanisms respon-
sible for TCA peeling have yet to be fully determined.
Previously, we showed that some specific growth factors includ-
ing platelet-derived growth factor (PDGF)-B are transiently pro-
duced by TCA-treated Pam212 murine keratinocytes and human
skin specimens after TCA application (25). The sources of PDGF
in human dermal wound repair are keratinocytes, as well as plate-
lets and monocytes. Platelet-derived growth factor stimulates tis-
sue fibroblasts around the wound to proliferate, to express the
appropriate integrin receptors and to migrate into the wound
space and presumably enhance wound closure by stimulating re-
epithelialization. In addition, in TCA-treated Pam212 murine
keratinocytes, expression of cytokines such as IL-1 and IL-10 were
also induced depending on TCA concentrations. In terms of cyto-
kines, proinflammatory cytokines (s IL-1) and anti-inflammatory
cytokines (e.g. IL-10) were both dose-dependently upregulated
after TCA treatment. These data suggest that the inflammatory
reaction after TCA treatment is well-balanced, resulting in a cos-
metically better outcome.
What role does activation of the SSRS, as described previously,
play in TCA peeling? Our study showed that POMC mRNA and
protein expression was specifically induced by TCA peeling with-
out induction of CRH expression in the epidermis. It is possible
that POMC expression is induced either directly by TCA or via
TCA
Epidermis
Prolifera on Inflamma on
SSRS
PDGF-B IL-1
TGF-β1 POMC
TGF-α IL-10
VEGF
MC-1R
Figure 3. Actions of TCA peeling on the epidermis. Supposed but uncertified
signalling pathways are indicated with dashed lines.
production of inflammatory cytokines such as IL-1a, and analysis
of the detailed mechanism is currently underway. At present, very
little is known about the biological activity of POMC itself. But in
the light of the overlap between the various antistress actions of
POMC-related peptides such as ACTH, MSH and b-endorphin,
POMC itself may also act similarly in terms of anti-inflammatory
action, immunosuppression, keratinocyte and fibroblast prolifera-
tion, and enhancement of skin turnover, other than melanocyte
stimulation. These actions may be involved in the regulation of
inflammation after TCA peeling and in good wound healing. We
have summarized the overall picture of the actions of TCA peeling
on the epidermis, including hypothetical actions, in Fig. 3, based
on an axis of the three regulatory pathways of wound healing;
proliferation, inflammation and SSRS activation.
Conclusion
We have outlined the HPA axis, an endocrine response system to
systemic stress, and the SSRS, its skin-specific counterpart and
have provided our own experimental data in a discussion of the
role played by SSRS in the active stress loaded to the skin by
chemical peeling as a specific rejuvenation treatment, especially by
TCA peeling as a model. Such a possible involvement of SSRS in
TCA peeling provides further insights on investigators of the
mechanisms of chemical peeling.
Conflicts of interest
The authors declare no conflict of interests.

References
1 Brodland D G, Cullimore K C, Roenigk R K.
J Dermatol Surg Oncol 1989: 15: 967–971.
2 Kimura A, Kanazawa N, Li H J et al. J Dermatol
2011: 38: 740–747.
3 Thorén P, Floras J S, Hoffmann P et al. Med Sci
Sports Exerc 1990: 22: 417–428.
4 Fukata J, Imura H, Nakao K. J Endocrinol Invest
1994: 17: 141–155.
5 Blalock J E. Physiological Rev 1989: 69: 1–69.
6 Slominski A T, Botchkarev V, Choudhry M et al.
Ann N Y Acad Sci 1999: 885: 287–311.
7 Kono M, Nagata H, Umemura S et al. FASEB J
2001: 15: 2297–2299.
8 Slominski A, Wortsman J, Mazurkiewicz J E et al.
J Lab Clin Med 1993: 122: 658–666.
9 Slominski A, Ermak G, Mazurkiewicz J E et al. J
Clin Endocrinol Metab 1998: 83: 1020–1024.
10 Slominski A, Baker J, Ermak G et al. FEBS Lett
1996: 399: 175–176.
11 Chakraborty A K, Funasaka Y, Slominski A et al.
Biochim Biophys Acta 1996: 1313: 130–138.
12 Cui R, Widlund H R, Feige E et al. Cell 2007:
128: 853–864.
13 Paus R, Botchkarev V A, Botchkareva N V et al.
Ann N Y Acad Sc 1999: 885: 350–363.
14 Slominski A, Ermak G, Hwang J et al. Biochim
Biophys Acta 1996: 1289: 247–251.
15 Lipton J M, Catania A. Immunol Today 1997: 18:
140–145.
16 Chakraborty A, Pawelek J. J Cell Physiol 1993:
157: 344–350.
17 Luger T A, Scholzen T, Brzoska T et al. Ann N Y
Acad Sci 1998: 840: 381–394.
18 Wakamatsu K, Graham A, Cook D et al. Pigment
Cell Res 1997: 10: 288–297.
19 Mika A, Romuald V. FRAGRANCE J 2005: 298:
35–38.
20 Bigliardi P L, Büchner S, Rufli T et al. J Recept
Signal Transduct Res 2002: 22: 191–199.
21 Bigliardi P L, Sumanovski L T, Büchner S et al.
J Invest Dermatol 2003: 120: 145–152.
22 Pritchard L E, White A. Endocrinology 2007:
148: 4201–4207.
23 Rousseau K, Kauser S, Pritchard L E et al. FASEB
J 2007: 21: 1844–1856.
24 Rakic L, Lapière C M, Nusgens B V. Skin Pharma-
col Appl Skin Physiol 2000: 13: 52–59.
25 Yonei N, Kanazawa N, Ohtani T et al. Arch Der-
matol Res 2007: 299: 433–440.

ª 2012 John Wiley & Sons A/S

10 Experimental Dermatology, 21 (Suppl. 1), 8–10