Arch Dermatol Res (2008) 300:397413
DOI 10.1007/s00403-008-0879-5
R EV IE W
The sebocyte culture: a model to study the pathophysiology
of the sebaceous gland in sebostasis, seborrhoea and acne
Christos C. Zouboulis Silke Schagen
Theodosios Alestas
Received: 10 May 2008 / Revised: 30 June 2008 / Accepted: 3 July 2008 / Published online: 9 August 2008
Springer-Verlag 2008
Abstract Acne is the most common skin disease which
aVects millions of people worldwide. Seborrhea and sebo-
stasis are major cosmetic problems but also lead occasion-
ally to diseases. This article summarizes the data of newest
research of sebostasis, seborrhoea and acne made possible
through the development of human and animal sebocyte
culture models.
Keywords Sebocytes Sebaceous gland
Skin physiology Sebostasis Seborrhea Acne
Treatment SZ95 sebocytes Hamster sebocytes
Rat sebocytes Mouse Sebocytes Hormones
Hormone receptor Sebocyte development
Propionibacterium acnes PPAR Cells
Cultured Lipids InXammation Drugs
C. C. Zouboulis, S. Schagen and T. Alestas contributed equally to the
article.
C. C. Zouboulis (&)
Departments of Dermatology, Venereology,
Allergology and Immunology, Dessau Medical Center,
Auenweg 38, 06847 Dessau, Germany
e-mail: christos.zouboulis@klinikum-dessau.de
C. C. Zouboulis S. Schagen
Laboratory for Biogerontology,
Dermato-Pharmacology and Dermato-Endocrinology,
Institute of Clinical Pharmacology and Toxicology,
Charit Universitaetsmedizin Berlin, Berlin, Germany
T. Alestas
Department of Dermatology, Andreas Sygros Hospital,
National and Capodistrian University of Athens,
Athens, Greece
Introduction
Acne is a disorder of certain pilosebaceous units, the seba-
ceous follicles. Each sebaceous follicle consists of a small
hair follicle, the hair bulge, the dermal papilla and an asso-
ciated relatively large sebaceous gland with a sebaceous
duct. Sebaceous glands acquire an intrinsically determined
morphology and pattern of behaviour during their develop-
ment which is modulated by several factors [134].
Sebocytes are the cells of the sebaceous gland, which
synthesize and accumulate lipid droplets. The biologic
activity of sebocytes is regulated by ligands of receptors
expressed in sebocytes, such as androgen and estrogen
receptors, peroxisome proliferator-activated receptors
(PPAR) and liver-X receptor (LXR) [37, 81], neuropeptide
receptors, retinoid and vitamin D receptors (reviewed in
[85, 115, 118]. The ligand-receptor complexes activate
pathways involving cell proliferation, diVerentiation, lipo-
genesis, hormone metabolism and cytokine and chemokine
release.
The pathogenesis of acne, the most common disease of
the sebaceous follicle, has currently been attributed to mul-
tiple factors such as increased sebum production, alteration
of the quality of sebum lipids, regulation of cutaneous ste-
roidogenesis, androgen activity, interaction with neuropep-
tides, exhibition of pro- and anti inXammatory properties,
follicular hyperkeratinization and the action of Propioni-
bacterium acnes (P. acnes) within the follicle [119, 131].
The amount and quality of acne research has signiW-
cantly increased in the last years by using such newly
emerged cell culture models and molecular techniques and
the results obtained have improved our understanding of
acne pathogenesis and the determination of targets for
future drug development. On the other hand, the use of ani-
mals in acne and sebaceous gland research has rapidly
123
398
reduced. Mammal sebocytes and sebocyte-like cells
(human, mouse, hamster and rat) and human sebaceous
gland cell lines (SZ95, SEB-1, Seb-E6E7) can be used in
monolayer cultures as models to study speciWc functions
involved in development, growth and diVerentiation of
sebaceous gland cells. Maintenance of these cells in certain
culture conditions has helped in investigating the physiol-
ogy of the sebaceous gland, including its changes in acne.
More complex culture systems, including three-dimen-
sional models, are under development.
Human sebaceous gland isolation and culture
Experimental sebaceous gland models are essential for a
better understanding of the pathophysiology of human skin
disorders and diseases involving the sebaceous gland, such
as sebostasis, seborrhoea and acne and for thorough devel-
opment of cosmetics and drugs and investigation of drug
pharmacokinetics. Data on sebaceous gland physiology ini-
tially emerged from the use of animal sebaceous gland
models. Animal models yield a higher number of sebocytes
than human sebaceous glands. Indeed, it is much easier to
isolate sebaceous glands and in greater numbers from ham-
ster skin specimens than from human skin biopsies. Ham-
ster sebaceous gland-derived cells have a lipid synthetic
capacity, although the components typical of human sebum
secretion, such as squalene and wax esters are not pro-
duced, conWrming their species-speciWcity [38]. On the
other hand, acne is exclusively a human disease. Since the
Wrst development of a human sebaceous gland cell line
(SZ95 sebocytes) [128], sebaceous gland research has
experienced a new era with numerous research groups
including sebocytes in their scope, whereas animal models
have almost been abandoned. Thus, the human sebocyte
culture is a unique tool for research on sebostasis, sebor-
rhoea, acne and other sebaceous gland-associated disorders.
Cultured sebocytes maintained properly preserve the major
Fig. 1 a Culture of human seba-
ceous gland. In the centre the
whole sebaceous gland is seen
and in the periphery extended
sebocytes are displayed. b Cul-
ture of human primary sebo-
cytes. Typical sebocytes of
diVerent maturation stages pre-
serving the major characteristic
of in vivo sebocytes
123
Arch Dermatol Res (2008) 300:397413
characteristic properties of human sebocytes in vivo and,
therefore, the human sebocyte culture model is closer to the
human in vivo situation [29, 117].
In 1966, Kellum [44] Wrst described the isolation of
sebaceous glands from human skin; however, his experi-
ments did not lead to the maintenance of viable sebaceous
glands in vitro. In 1982, Karasek and Charlton [42]
reported in an abstract the cultivation of human sebocytes,
obtained by dermal slices rich in sebaceous gland tissue, as
a model for the investigation of the human sebaceous
glands, but provided not extended publication of their
work. Their technique has been later modiWed and used by
Doran et al. [19] in several experiments and is occasionally
still used nowadays by some investigators. In 1986, Kealey
et al. [43] and in 1989, Xia et al. [110] introduced the main-
tenance of the sebaceous gland ex vivo and a reproductive
model for the cultivation of sebaceous gland cells in vitro,
respectively (Fig. 1). Several modiWcations of the technique
of Xia et al. regarding sebocyte isolation and cultivation
have further facilitated the cultivation of human sebocytes
[110].
However, human sebocytes are predisposed to diVerenti-
ate by accumulating neutral fat droplets until they burst and
die. Therefore, adequate cell amounts for large scale exper-
iments can only be obtained from multiple donors, while
prolonged experiments are hindered by the short life span
of the cells. Normal human sebocytes can only be grown
for three to six passages [114], to overcome this problem,
Zouboulis et al. [128] have cultured human facial sebocytes
from a 87-year-old woman and transfected them with the
simian virus-40 large T antigen. The resulting immortalized
cell line has been cloned, certain clones have been charac-
terized and it was termed SZ95. The SZ95 sebaceous gland
cell line is now protected by national and international pat-
ents as well as priority submissions (WO0046353,
AT319813T, AU770518B, CA2360762, CN1344314T,
DE19903920, DK1151082T, EP1151082, HU0200048,
IL144683D, JP2002535984T, KR689120, PL194865,
Arch Dermatol Res (2008) 300:397413 399

US2002034820 and others). Several studies have shown

that SZ95 sebocytes retain major characteristics of normal
human sebocytes, such as progressing diVerentiation with
increasing cell volume and lipid synthesis, expression of
markers of sebaceous lineage and terminal sebocyte diVer-
entiation [114], such as keratin 7 and epidermal membrane
antigen (EMA) respectively [114, 125] and can subse-
quently undergo apoptosis [109]. They also express charac-
teristic origin- and function-speciWc proteins of human
sebaceous glands and exhibit expected biological responses
to androgens and retinoids [109, 128].
In 2003, Thiboutot et al. [95] have applied the transfec-
tion system administered by Zouboulis et al. [128] to
develop a second immortalized human sebaceous gland cell
line, termed SEB-1. SEB-1 was established from sebaceous
glands of normal skin of the preauricular area of a 55-year-
old male [95]. Like SZ95 sebocytes, SEB-1 sebocytes also
express characteristic sebaceous gland proteins and their
cytoplasm induces oil red O-positive lipid droplets. In gene
array studies, genes characteristic for the sebaceous gland
and such involved in lipid and steroid metabolism were
expressed in SEB-1 sebocytes.
A third immortalized sebaceous gland cell line, Seb-
E6E7, has been generated from adult human facial skin fol-
lowing a facelift procedure [52]. Human sebocytes were
immortalised by introduction of HPV16 E6 and E7 genes.
Seb-E6E7 sebocytes were transduced by co-culture with
mitomycin C-treated packaging cells in the presence of
3T3-J2 cells. Seb-E6E7 sebocytes, like SZ95 sebocytes,
express both K7 and involucrin. In Wrst experiments, Seb- Fig. 2 Model follicle presenting the development of sebocytes, der-
E6E7 seems to respond to chemicals in a similar manner mal papilla cells and epidermal cells (keratinocytes) [77])
with SZ95 sebocytes despite their diVerent transfection
methods. Among them -catenin and lymphoid enhancer factor-1
seem to be of major importance [67, 94]. The level of -
catenin regulates lineage selection by stem cell progeny in
Development of sebaceous glands and sebocytes mammals. High levels of -catenin stimulate the formation
of hair follicles and low levels that of epidermis and seba-
Epidermal progenitor cells give rise to multiple skin lin- ceous glands. Intracellular signalling molecules like tran-
eages: hair follicle, sebaceous gland and the overlying scription factor 3 (Tcf3) and lymphocyte-enhancing factor-
interfollicular epidermis [77]. The multipotent stem cells 1 (Lef-1), a DNA binding molecule, control diVerentiation
reside in the bulge region of the hair follicle. These cells lineage [67]. Overexpression of Lef-1 blocks -catenin sig-
transform into the epidermis (epidermal keratinocytes) as nalling, which may lead to spontaneous sebaceous tumours.
well as its associated structures sebaceous gland (sebo- Lef-1 and Indian hedgehog (IHH) seem to cooperate to
cytes) and cells of the hair follicle (follicular keratinocytes) control proliferation and diVerentiation of sebocyte progen-
(Fig. 2). itors (Fig. 3). Sonic hedgehog (SHH) is a signalling path-
Many of the molecules which regulate epidermal self- way, which is important in embryos and adults for the
renewal and diVerentiation have now been identiWed. Sev- regulation of progenitor cells of hair lineage diVerentiation
eral molecular networks and signalling pathways are and proliferation. SHH, which is a controlled member of
important in balancing epidermal growth and diVerentia- the Wnt signalling pathway, is required for terminal diVer-
tion. Key molecules include nuclear factor
B (NF-
B), entiation of hair lineage [25, 64]. Inhibition of Wnt target
Wnt/-catenin, sonic hedgehog/patched, p63, 14-3-3, - genes using a dominant negative Lef-1 promotes sebocyte
catenin and 1-integrin [106]. The signals for sebocyte development, while inhibiting diVerentiation of hair lineage
development have currently been partially understood. [58, 66].

123
400
stem cell
-catenin N LEF-1
N LEF-1
transit cells
IHH SHH
follicular epidermal
sebocyte
keratinocyte keratinocyte
Fig. 3 Model of interactions between
NLef-1, -catenin and hedge-
hog signalling in epidermal stem and progenitor cells. The high level
of -catenin is necessary to develop follicular keratinocytes,
NLef-1
blocks -catenin for development of epidermal keratinocytes; involve-
ment of IHH leads to the development of sebocytes (from [67])
The Wndings of Allen et al. [6], who were able to
develop sebaceous glands at the food pad of the mouse,
raise the possibility that sebocyte fate is governed by the
relative level of stimulatory (hedgehog) and inhibitory
(Wnt) signals acting on multipotent progenitors. The
hedgehog pathway and other molecules are implicated in
sebocyte development, including c-Myc, PPAR and cyclo-
oxygenase (COX)2 [6]. Activation of c-Myc stimulates epi-
dermal proliferation without depleting label retaining cells
and induces diVerentiation of sebocytes within the interfol-
licular epidermis [10].
Sebum and sebaceous lipids
One of the main functions of sebocytes is synthesis of lipids
(Fig. 4). Human sebum, a mixture of sebaceous lipids and
cell debris, is composed of glycerides, wax esters, squalene,
cholesterol and cholesterol esters and free fatty acids. It has
unique properties; squalene and certain fatty acids are only
synthesized by sebocytes [72]. Squalene is the linear inter-
mediate in cholesterol biosynthesis. While in other tissues
squalene is quickly converted to lanosterol and Wnally to
cholesterol, squalene produced in sebaceous cells is not
converted to cholesterol. Squalene is produced by the
fusion of two molecules of farnesyl pyrophosphate through
the action of squalene synthase. Wax esters biosynthesis is
increased in human sebaceous gland cells in comparison
123
Arch Dermatol Res (2008) 300:397413
Fig. 4 Fluorescence microscopy picture of nile red stained SZ95 seb-
ocytes: Polar lipids (red) such as phospholipids containing organelles,
membrane lipids are measured by 540 nm excitation and emission
620 nm Wlters and for neutral (unpolar) lipids such as triglycerides,
glycerides and wax esters are visualised at 485 nm excitation and
528 nm emission Wlter (orange)
with other species [72]. Fatty acid and fatty alcohols are the
natural and direct precursors of wax esters. In addition, seb-
ocytes synthesize in vitro less squalene, wax esters and
cholesterol esters than in vivo, but their lipogenesis can be
upregulated, e.g. by treatment with linoleic acid (LA) or
arachidonic acid (AA) [82, 127]. The lipogenic factors
CCAAT/enhancer binding protein transcription factors,
galectin-12, resistin and sterol response element binding
protein 1 (SREBP1) were detected in SZ95 sebocytes [13,
36]. SREBP1 has been shown to increase the transcription
of human squalene synthase in the liver of transgeneic mice
and SREBP1 was shown to be increased in SEB-1 sebo-
cytes in response to insulin and insulin-like growth factor 1
(IGF-1) [91]. Squalene synthase mRNA is express in SZ95
sebocytes [52] and squalene may serve as a marker for ter-
minal sebocyte diVerentiation [127].
Ge et al. [28] have identiWed
6-desaturase/fatty acid
desaturase 2 (FADS2) as the major fatty acid desaturase in
human sebaceous glands, which sebocytes use to synthesize
very long chain fatty acids. Desaturase add double bonds
between carbons 6 and 7 of precursor to let sebocytes gen-
erate the polyunsaturated fatty acids -linolenic and steari-
donic acid, respectively (Table 1). Low levels of linoleic
acid have been observed in skin surface lipids of acne
patients [21]. Linoleic acid undergoes peroxisomal -oxi-
dation to AA and other fatty acids, whereas the ability of
sebocytes to synthesize wax esters correlates with the level
of -oxidation in these cells. It seems that -oxidation of
linoleic acid is sebocyte-speciWc and is associated with
their diVerentiation [72].
Palmitic acid
6-desaturation yields the monounsaturated
sapienic (palmitoleic) acid. Sapienic acid and sebaleic acid
(18:2, 5, 8), are the predominant fatty acids, which are unique
Arch Dermatol Res (2008) 300:397413
Table 1 Sebaceous fatty acids
Precursor
and their synthesis
Linoleic acid
(6, C18:2, 9,12)a
-linolenic acid
(3, C18:3, 9,12,15)a
Palmitic acid (16:0)
a
Essential fatty acids
b
mRNA and protein expression
detected in SZ95 sebocytes [
Stearic acid (18:0)
30, 73]
in humans to other hair bearing mammals [28, 64, 72, 93].
Sapienic acid of human sebum exhibits antibacterial activity
against gram-positive bacteria, like P. acnes [108]. P. acnes
release lipases that initiate sebum metabolism, whose prod-
ucts have been suggested to induce inXammation [11].
In adition to FADS2, human facial sebocytes also
express functional stearoyl CoA desaturase (SCD), the
most upstream enzyme in steroidogenesis [30, 36]. The
microsomal enzyme SCD is an iron-binding 41 kDa protein
of 355 amino acids with six predicted transmembrane
domains. It catalyses
9-desaturation of long-chain unsatu-
rated fatty acids, leading to the biosynthesis of palmitoleic
acid and oleic acid as its major products.
Increased squalene peroxidation and reduction of lino-
leic acid concentration lead to hypercorniWcation of ductal
keratinocytes [15, 21, 93]. The free fatty acid fraction of
sebum has been considered to participate in the inXamma-
tory process of acne. Feeding of the AA synthetic pathway
by linoleic acid is the Wrst step into production of proin-
Xammatory cyclooxygenase products. In addition, squalene
peroxide has been shown to stimulate the proliferation of
HaCaT keratinocytes in vitro [70].
Nagai et al. [60] detected that cultured cloned hamster
and rat primary sebocytes secrete membrane vesicles, deW-
ned as sebosomes, containing squalene condensed lipid par-
ticles and enriched with histone H3, a cationic protein.
These sebocyte vesicles suggest a novel function of these
cells, which is associated with the secretion of antibacterial
proteins and sterol regulation. These molecules may protect
the skin surface. On the surface of intracellular lipid drop-
lets in diVerentiated hamster sebocytes in vitro perilipin A
expression has been identiWed [4]. Perilipin is a mammal
protein, which envelops lipid droplets in fat cells and pro-
tects them from the fat burning enzyme hormone-sensitive
lipase. Additionally, the fatty acid transport protein 4
(FATP4) is strongly expressed in sebaceous glands. This
trans-membrane protein enhances the uptake of long chain
fatty acids into the cells [84].
Sebaceous lipids can be visualized with the nile red/oil
red stainings [33] (Fig. 4). Squalene, wax esters, triglycer-
ides and free fatty acids have been detected in vitro by thin
401
Products Enzyme
-linolenic acid (6, C18:3, 8,11,14) Fatty acid
desaturase 2 (FADS2)b
stearidonic acid (3, C18:4, 6,9,12,15) FADS2
stearidonic acid (3, C18:4, 6,9,12,15) FADS2
Sapienic acid (16:1, 6) FADS2
Palmitoleic acid (C16:1, 9) Stearoyl CoA
desaturase (SCD)b
Oleic acid (C18:1, 9) SCD
layer chromatography [128], being a sign of mature sebo-
cytes. SZ95 sebocyte treatment with AA leads to lipid
accumulation and cell apoptosis. Terminal diVerentiation
and apoptosis are two diVerent programmed cell events, but
both are followed by cell death. Lipid synthesis in sebo-
cytes is a part of their terminal diVerentiation. It begins by
accumulation of lipid droplets in cytoplasm which is fol-
lowed by cell burst and death. Apoptosis is involved in this
process, shown by Wndings of nuclear fragmentation and
bcl-2 reduction in cultured SZ95 sebocytes [109]. AA also
up-regulates the secretion of interleukin (IL)6 and IL8 in
sebocytes [5]. AA is metabolized by 5-lipoxygenase to leu-
kotriene (LT) B4, a ligand of PPAR, and upregulates lipo-
genesis in human sebocytes
The regulation of lipogenesis by all-trans retinoic acid
(atRA) and androgens in hamster sebocytes is similar to the
regulation in human sebocytes [128]. Epidermal growth
factor and 1,25-dihydroxyvitamin D3 (Vit. D3) act as sup-
pressors in the regulation of lipogenesis in hamster sebo-
cytes in vitro [83]. Epidermal growth factor, transforming
growth factor (TGF)- and Wbroblast growth factor have
mitogenic activity on hamster sebocytes and the two latter
act as anti-lipogenic factors. Moreover, it is likely that the
formation of intracellular lipid droplets is independent of
cell proliferation in hamster sebocytes [3].
Expression of characteristic proteins / hormones
of human sebaceous gland cells and their biological
activity
Human skin is an endocrine organ which synthesizes hor-
mones itself. Hormones are biochemically active messen-
ger compounds, which coordinate activity in diVerent cell
types in multi-cellular organisms. Skin cells are able to
metabolize, activate and inactivate hormones. Human sebo-
cytes produce corticotrophin-releasing hormone (CRH),
androgens, estrogens, atRA, cortisol, vit. D3 and eicosa-
noids [115, 119, 134]. Sebocyte proliferation, diVerentia-
tion and lipid synthesis (accumulation of neutral lipids) are
controlled by associated complex endocrine pathways [35].
123
402
Hypothalamus secretes CRH in a cicardian manner;
seven to ten times a day (in stress situations even more
often), aVecting the pituitary gland. The latter synthesizes
propiomelanocortin (POMC) and decomposes it into corti-
cotropin (ACTH), -endorphin as well as -melanocyte-
stimulating hormone (-MSH). Interestingly, CRH is
expressed in human sebocytes in vivo and in culture [49,
129] and directly upregulates 3-
54-hydroxysteroid
dehydrogenase (3-HSD), the enzyme that converts dehy-
droepiandroestrone (DHEA) to testosterone [24]. CRH
stimulates the synthesis of neutral (sebaceous) lipids and of
IL6 and IL8 in human sebocytes [49, 129]. CRH expression
is upregulated in acne-involved sebaceous glands in vivo
when compared with non-involved sebaceous glands of
acne patients and sebaceous glands of healthy individuals
[26].
Androgens aVect several functions of human sebaceous
glands in vivo, such as stimulation of cell proliferation
and diVerentiation, including lipid synthesis [133]. In cell
culture, androgens promote sebocyte proliferation [24]
and also lipogenesis under the presence of PPAR ligands
[83]. Estrogens, cortisol and atRA downregulate diVeren-
tiation, increase cell proliferation and decrease the intra-
cellular accumulation of neutral lipids [14, 36, 109]. In
addition, they regulate CD14 and Toll-like receptors
(TLR) [68]. The eVect of estrogens can be explained by
inhibition of gonadotropin secretion or by enhancement
of testosterone binding to its binding globulin. Cortisol
also inXuences lipid metabolism, wound healing and
relieves stress [115, 118].
Vit. D3 exhibits various cellular functions including anti-
proliferative eVects on SZ95 sebocytes in the logarithmic
phase of cell growth and diVerentiation [87]. In addition, it
decreases lipid synthesis in hamster sebocytes [83].
Eicosanoids such as prostaglandins (PG), prostacyclins
and LT are AA derivatives. Eicosanoid synthesis can
induce inXammatory signals in human sebocytes [5].
Hormone receptors and their properties in human
sebocytes
Sebocytes express receptors for peptide hormones, neuro-
transmitters, which are mostly arranged on the cell surface,
and for steroid and thyroid hormones, which are found in
the cytoplasm or nuclear compartment.
Three of four groups of peptide hormone and neurotrans-
mitter receptors are represented in human sebocytes
(Table 2). To the Wrst so-called serpentine or seven trans-
membrane domain receptor group belong
CRH recepror (CRH-R)1 and 2, whereas CRH-R1 is
more abundant and seems to regulate CRH activity [49,
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Arch Dermatol Res (2008) 300:397413
129]. Through binding to CRH-R1, CRH and urocortin
reduce sebocyte proliferation, CRH upregulates
54-
3-hydroxysteroid dehydrogenase expression, synthe-
sis of neutral lipids and IL6 and IL8 release;
melanocortin-1 and 5 receptors (MC-1R and MC-5R),
which bind -MSH and are located at the cellular sur-
face of sebocytes. MC-1R regulates inXammation in
SZ95 sebocytes [8] and exhibits a stronger expression
in acne-involved sebaceous glands [27]. The expres-
sion of MC-5R is weaker than that of MC1-R but has
been shown to be a marker of human sebocyte diVeren-
tiation, since it is expressed in diVerentiated, lipid-con-
taining sebocytes, only [111].
-opiate receptors (OPR), which bind -endorphin. -
endorphin stimulates lipogenesis and speciWcally
increases the amount of C16:0, C16:1,C18:0, C18:1
and C18:2 fatty acids to an extent similar to linoleic
acid in sebocytes [9].
VPAC receptors, which bind vasoactive intestinal
polypeptide (VIP), receptors for neuropeptide Y (NY)
and calcitonin gene-related peptide (CGRP) [89]. NY
activates cytokine synthesis. CGRP is often co-local-
ized with substance P [92].
Cannabinoid receptors (CR) 1 and 2 are expressed in
SZ95 sebocytes and sebaceous glands [18]. CBR1 was
found in the diVerentiated sebocytes and CBR2 in the
undiVerentiated cells, whereas endocannabinoids inXu-
ence sebocyte diVerentiation via CR2.
Histamine 1 receptor, with binds with histamine and
regulates squalene synthesis [73]. Antihistaminics,
ligands of histamine 1 receptor reduced squalene syn-
thesis in SZ95 sebocytes.
The IGF-I receptor belongs to the second group, the single-
transmembrane domain receptors, that harbor intrinsic tyro-
sine kinase activity, is expressed on SZ95 sebocyte cell sur-
face and can be activated by IGF-I and high concentrations
of insulin [55, 129]. IGF-I ampliWes lipid accumulation in
SZ95 sebocytes in a dose dependent manner. The activation
of the IGF-I receptor induced lipogenesis in SEB-1 sebo-
cytes by SREBP-dependent and independent pathways
[91]. IGF-I also stimulates proliferation and diVerentiation
of rat preputial gland cells, which resemble sebocytes,
especially in combination with GH [16].
The third group, which is functionally similar to the sec-
ond group does not possess intrinsic tyrosine kinase activity
but appear to function through interaction with soluble
transducer molecules which do possess such activity. In
human sebocytes, they are represented by the GH receptor
[129], whose regulation by GH up-regulates sebocyte
diVerentiation and augments the eVect of 5-dihydrotestos-
terone (DHT) on sebum synthesis [16].
 Table 2 Hormone receptors expressed in human sebocytes
Receptors Natural ligands EVect on cultured sebocytes References

Peptide hormone and neurotransmitter receptors

Serpentine (seven transmembrane domain) receptors
Corticotropin-releasing hormone (CRH) CRH, urocortin # proliferation, "
54 3-hyroxysteroid [49, 129]
receptors 1 and 2 (CRH-R1 > CRH-R2) dehydrogenase (CRH), " lipid synthesis (CRH),
" IL6 and IL8 release (CRH)
Melanocortin-1 and 5 receptors -melanocyte-stimulating # IL1-induced IL8 synthesis (MC-5R), [8, 111]
(MC-1R and MC-5R) hormone (-MSH) diVerentiation marker (MC-1R)
Arch Dermatol Res (2008) 300:397413

-opiate receptors (OPR) -endorphin # EGF-induced proliferation, " lipid synthesis [9]
VPAC receptors vasoactive intestinal popypeptide (VIP), " IL6 and IL8 release (neuropeptide Y) [89]
receptors for neuropeptide Y and
calcitonin gene-related peptide (CGRP)
Cannabinoid receptors (CR1 and CR2) cannabinoid InXuence of sebocyte diVerentiation (CR2) [18]
Histamine receptor 1 histamine Regulation of squalene synthesis [73]
Single-transmembrane domain receptors with
intrinsic tyrosine kinase activity
Insulin-like growth factor-I (IGF-I) receptor IGF-I, insulin " lipogenesis [16, 55, 90, 129]
Single-transmembrane domain receptors without
intrinsic tyrosine kinase activity
Growth hormone (GH) receptor GH " diVerentiation, " DHT eVect on lipogenesis [16, 129]
Nuclear receptors
Steroid receptors
Androgen receptor Testosterone, " proliferation (with PPAR ligands " lipogenesis) [2, 23, 24, 53, 115, 118, 128]
5-dihydrotestosterone (DHT)
Progesterone receptor Progesterone ? [74, 133]
Thyroid receptors
Estrogen receptors (ER-- and -) 17-estradiol " polar lipid production [56, 74, 97, 133]
Retinoic acid receptors (RAR- and -) all-trans retinoic acid (atRA) # proliferation [75, 105]
Retinoid X receptors (RXR-, > PXR-, -) 9-cis RA Regulate lipogenesis (?) [46, 105]
Vitamin D (Vit. D) receptor Vit. D3 Regulates cell proliferation, cell cycle, [76, 88]
lipid content and IL6 and IL8 secretion
Peroxisome proliferator-activated receptors Linoleic acid (RRAR/), " lipogenesis, " prostaglandin E2 release, [5, 53, 84, 112]
(PPAR, PPAR > PPAR) leukotrien-B4 (LTB4) " IL6 release, " cycloxygenase 2 (COX2) synthesis
Liver X receptors (LXR and -) 22(R)-hydroxycholesterol # proliferation, " lipogenesis, # COX2 and inducible [37, 81]
nitric oxide synthase

123
403
404
The nuclear receptors are soluble molecules and employ
transcriptional regulation as a means of promoting their
biological eVects. Thus, though some receptors are com-
partmentalized in the cytoplasm while others are deWned to
the nucleus, they all operate within the nucleus chromatin
where they bind a hormone-speciWc hormone response
element. These receptors are expressed in human sebo-
cytes and can be grouped into two major subtypes based on
shared structural and functional properties.
From the Wrst group, the steroid receptor family, the
androgen receptor (AR) and the progesterone receptor (PR)
are present in human sebocytes, in basal and early diVeren-
tiated ones [24, 115, 118, 133].
AR is stabilized und upregulated by ligand binding; its
downregulation reduces sebocyte proliferation [23].
Five intracellular enzymesall of them expressed in
sebocytes [24]are involved in activationbefore
binding to ARand inactivation of androgens. DHEA-
sulfate is metabolized by SCD to DHEA. DHEA and
androstosterone are converted to testosterone and later
to DHT by 5-reductase [24, 12]. Sebocyte studies of
Akamatsu et al. and Zouboulis et al. showed a dose
dependent induction of sebocyte proliferation by tes-
tosterone treatment [2] and no eVect on lipid stimula-
tion [128]. Investigations by RosenWeld et al. and
Makrantonaki et al. proved that the eVect of androgens
on sebaceous lipids is mediated by PPAR ligands [53,
78].
PR was found in nuclei of basal sebocytes of sebaceous
glands [74, 133].
From the second group, the thyroid receptor family, the
following receptors are expressed in human sebocytes:
Estrogen receptors (ER; - and -isotypes) [74, 97,
133]. ER is expressed in basal and partially diVerenti-
ated sebocytes. ER is expressed in basal and early
diVerentiated sebocytes [98]. One of the natural estro-
gens, estradiol, is created by oxidative reduction of 4-
androsten-3, 17-dion, like testosterone. Treatment of
sebocytes with 17-estradiol showed an eVect on polar
lipid production but no stimulating eVect on neutral lip-
ids [56]. Other previous in vitro data indicated that
estrogens may have an inXuence on the biological
activity of sebaceous glands [34, 35].
Retinoic acid receptors (RAR; isotypes  and ) and
retinoid X receptors (RXR; isotypes , , ) [75, 105].
RAR and  and RXR are the predominant retinoid
receptors in human sebocytes, RAR regulates cell pro-
liferation [105]. The natural ligands for RAR and RXR
are atRA and 9-cis retinoic acid. 13-cis retinoic acid
(13cRA) inhibits proliferation in SZ95 sebocytes,
whereas 13cRA was found to be metabolized intracel-
lularly to the RAR ligand atRA [105]. RXR agonists
123
Arch Dermatol Res (2008) 300:397413
are stimulating sebocyte diVerentiation and prolifera-
tion. The RXR agonist rexinoid in combination with
the speciWc PPAR agonists, WY 14643, troglitazone
and cabaprostacyclin aVected diVerentiation and
growth in cultured primary sebocyte-like rat preputial
cells [46].
Vitamin D receptor (VDR) [76]. SZ95 sebocytes also
express vitamin D-25-hydroxylase (25OHase), 25-
hydroxyvitamin D-1-hydroxylase (1OHase), and 1,
25-dihydroxyvitamin D-24-hydroxylase (24OHase)
[88]. Vit. D3 induces time- and dose-dependent modu-
lation of cell proliferation, cell cycle regulation, lipid
content and IL6 and IL8 secretion by cultured sebo-
cytes. RNA expression of VDR and 24OHase was
upregulated along with vit. D3 treatment.
PPAR [5, 53, 84]. PPAR and  are the predominant
PPAR subtypes in human sebocytes. PPAR are present
in mitochondria, peroxisomes and microsomes of sebo-
cytes and regulate multiple lipid metabolic genes.
Liver X receptors (LXR, - and - isotypes) [37, 81].
SZ95 sebocytes express both receptors at the mRNA
and protein levels. The application of natural [22(R)-
hydroxycholesterol] or synthetic ligands signiWcantly
inhibited proliferation and increased lipogenesis [37,
81]. The expression of known LXR targets, such as
fatty acid synthase and SREBP1, was induced by the
synthetic LXR ligand TO901317, which also
decreased the expression of COX2 and inducible
nitric oxide synthase that was induced by LPS treat-
ment [37].
The vanilloid receptor (VR) belongs to the transient ion
channels and is expressed in diVerentiating sebocytes [99]
VR ligand capsaicin was shown to reduced SZ95 sebocyte
proliferation.
The nuclear PPAR regulate sebaceous lipogenesis
PPAR require heterodimerization with RXR for binding to
DNA as ligand activated transcription factors that regulate
the expression of target genes containing peroxisome pro-
liferator response elements (PPRE) in their promoters.
PPAR have been shown to regulate multiple lipid metabolic
genes in peroxisomes, microsomes and mitochondria by
activating PPRE [32, 86]. Each of the PPAR subtypes are
expressed in rat preputial sebocyte-like cells, SZ95 and
SEB-1 immortalized cell lines and human sebaceous glands
at the mRNA and protein levels [13, 20, 79, 103] (Table 3).
Moreover, Chen et al. [13] and Alestas et al. [5] demon-
strated that all PPAR subtypes are present in sebaceous
glands and in the pilosebaceous ducts of healthy, acne
involved and acne uninvolved skin.
Arch Dermatol Res (2008) 300:397413 405

Table 3 Peroxisome proliferators activated receptor ligands in rodent lococci and diphtheroid rods on the skin surface and P.
and human sebocytes acnes in the infundibulum of the sebaceous glands. The dis-
PPAR PPAR ligands tribution and density of the Xora is dependent on age and
isotype environmental factors such as sebum secretion, occlusion,
Rodent sebocytes Human sebocytes
temperature and humidity. Antimicrobials may reduce the
 Synthetic ligands: Synthetic ligands: density of the skin resident Xora, but they cannot com-
WY WY, GW 7647 pletely eliminate it.
Natural ligands: GW 0742, GW 2433 P. acnes, gram positive bacteria, have been suggested to
LA, cPGI2 Natural ligands:
AA, LTB4 play an important role in pathogenesis of acne but the exact
/ Synthetic ligands: Synthetic ligands:
mechanism is still unclear. In 2004, Bruggemann [11] et al.
GW 0742, GW 2433 identiWed the complete genome sequence of P. acnes.
Natural ligands: Natural ligands: Although in some studies P. acnes numbers are higher in
LA, cPGI2 LA acne patients than in healthy individuals, other studies
 Synthetic ligands: Synthetic ligands: found no diVerence between the numbers of P. acnes in
TRO, BRL GW 4148, CIG, BRL,
Natural ligands: pioglitazone
aVected and non-aVected follicles [48]. P. acnes produces

15-PGJ2 Natural ligands: extracellular enzymes, such as lipases, proteases and hyal-
15-HEFE, PGJ2,
15-PGJ2 uronidase, which may participate in acne inXammation.
LA Linoleic acid, AA arachidonic acid, LTB4 leukotriene B4, WY Wy- These enzymes lead to activation of epithelial cells by pro-
14643, CIG ciglitazone, cPGI2 carbaprostacyclin, TRO troglitazone, 15- inXammatory signals. They initiate local interactions
HETE hydroxyeicosatetraenoic, BRL rosiglitazone, PG prostaglandin between bacterial products and inXammatory cells, such as
macrophages and leucocytes [7, 21, 40].
PPAR ligands induce lipogenesis in human sebocytes [5, Keratinocytes and sebocytes may be activated by P.
13, 53, 103, 109], whereas PPAR expression decreases with acnes via toll-like receptors (TLR), CD14 and CD1 mole-
progressing sebaceous diVerentiation in normal but not in cules [1, 31, 47, 61]. TLRs are transmembrane proteins
acne skin [5]. The most predominant PPAR in human seba- capable of mediating responses to pathogen-associated
ceous glands are RRAR and 1 [5]. PPAR seems to acti- molecules conserved among microbes [45]. Pilosebaceous
vate -oxidation of fatty acids, regulate inXammation and be follicles in acne lesions are surrounded with macrophages
associated with sebocyte diVerentiation. While stimulating expressing TLR2 on their cell surface. TLR2 activation
inXammation and sebaceous lipogenesis, AA only induces a leads to NF-kB triggering and thus production of cytokines/
reduction of PPAR mRNA expression in cultured SZ95 chemokines, phenomena which are observed in acne
sebocytes [5]. PPAR regulates the late stages of sebocyte lesions [41]. Furthermore, P. acnes induces IL8 and IL12
diVerentiation and stimulate sebaceous lipid synthesis [17]. release from TLR2 positive monocytes [45].
PPAR is suggested to activate sebocyte proliferation and Human SZ95 sebocytes express TLR2, TLR4, TLR6 and
lipogenesis [103]. PPAR is also involved in oxidative CD14 [30]. Upon stimulation with lipopolysaccharide
stress mediated PGE2 production and induces COX2 expres- (LPS), peptidoglycan or lipoteichoic acid SZ95 sebocytes
sion in SZ95 sebocytes [112]. PPAR is expressed and acti- release cytokines by TLR- and CD14-dependent mecha-
vated in SZ95 sebocytes by UVB irradiation. After sebocyte nisms [61]. Interestingly, the diVerent phylogenetic cluster
treatment with PPAR agonists increased COX2 mRNA and P. acnes isolates exhibited phenotypic diVerences when co-
protein expression and PGE2 production were observed incubated with SZ95 sebocytes [61]. SZ95 sebocyte treat-
[112]. By pre-treatment with a PPAR antagonist the eVec- ment with P. acnes isolates signiWcantly increased the num-
tiveness of PPAR agonists was blocked. ber of viable cells. P. acnes and LPS induced -defensin-2
Hamster sebocytes also express PPAR and PPAR Tert- and catelichidin expression in SZ95 sebocytes [50, 61].
buthylhydroperoxide (TBH) is a lipid soluble oxidant P.acnes also enhances the expression of IL8 andTNF but
which activates PPAR1 [4]. PPAR and 1 ligands induced not IL1 [60]. The latter is upregulated by LPS [68].
transcriptional augmentation of perilipin A expression Expression of IL6 and IL8 is enhanced in acne-inolved
along with an increase in levels of triacylglycerol in lipid sebaceous glands as detected by immunohistochemistry [5].
droplets in hamster sebocytes [4]. Some P. acnes-derived ligands of TLR have been identi-
Wed: GroEL, DnaK or lipoglycans may act as such ligands
[22, 31, 107].
P. acnes eVects on sebocytes On the other hand, Georgel et al. [30] observed the exis-
tence of a regulated, lipid-based antimicrobial eVector path-
Healthy human skin is colonized by non-pathogenic micro- way in mammals and suggest new approaches in the
organisms. Bacteria isolated are coagulase negative staphy- treatment or prevention of infections with gram positive

123
406
bacteria. A TLR2-dependent transcriptional activation of
SCD occurs during gram-positive bacteria exposure. SCD
is responsible for the biosynthesis of speciWc lipids with
antimicrobial activity, such as the monounsaturated fatty
acids palmitoleic and oleic acids. Since acne is an inXam-
matory but not an infectious disease [116], a new role of P.
acnes and other skin commensals has been suggested: in
contrast to the current opinion their presence may be skin
protective and required for a permanent low level of innate
immunity activation to defend skin from acute pathogen
attacks.
InXammation mechanisms in sebocytes
Until now the knowledge regarding the molecular mecha-
nisms of acne inXammation is not completely investigated.
Trivedi et al. demonstrated that 211 genes in lesional skin
biopsies of acne patients are up-regulated and 18 genes are
down-regulated compared to normal human skin biopsies.
Among the up-regulated genes are such ones involved in
inXammatory and extracellular matrix modelling pathways.
They include matrix metalloprotease 1 and 3, IL8, -defen-
sin 4 and granzyme B [104]. NF-
B and activator protein-1
are active in acne lesions with consequent increase in their
target gene expression, such as inXammatory cytokines,
enzymes and matrix-degrading metalloproteases (MMP)
[41]. SZ95 sebocytes in culture secrete pro MMP-2 and pro
MMP-9 [71].
InXammation is characterized by action of active lipid
mediators, such as LT, PG and 15-hydroxyeicosatetraenoic
acids. These molecules are synthesized from AA or lino-
lenic acid by the enzymes COX and lipoxygenase (LOX).
COX, a prostaglandin endoperoxide H synthase, is a rate
limiting enzyme complex for the production of PG. Both
COX isozymes, COX1 and COX2, are expressed in human
sebocytes in vitro and moreover, COX2 expression is selec-
tively up regulated in acne-involved sebaceous glands in
vivo [5]. In hamster sebocytes, COX2 expression is mea-
surable only [38]. The PG biological pathway plays an
important role in sebaceous gland cells.
PG overtakes regugulation of cellular function under
physiological and pathological conditions. PG are associ-
ated with sebaceous gland development and sebaceous
lipogenesis. In the augmentation of lipid droplet formation
in hamster sebaceous glands 15-desoxy-
12,14-PGJ2 (15d-
PGJ2) is a crucial factor by increasing triglycerol synthesis
in sebocytes. COX2 inhibitors reduce PGF2 and PGE2 lev-
els but increase 15d-PGJ2 production and triacylglycerol
synthesis. Additionally the synthesis of 15d-PGJ2 is medi-
ated by COX2 and cytochrome P-450 dependent pathways
[39]. Zhang et al. [113] demonstrated that the production of
PGE2 is induced by platelet-activating factor receptor
123
Arch Dermatol Res (2008) 300:397413
(PAF-R) activation and mediated by COX2 expression.
PAF-R seems to be involved in regulating the expression of
inXammatory mediators, including COX2, PGE2, and IL8.
LT are potent proinXammatory mediators and neutrophil
attractants. SZ95 sebocytes express LOX and LTA4 hydro-
lase at protein and mRNA levels. These enzymes are essen-
tial for the formation of LTB4. When SZ95 sebocytes were
treated with AA, LTB4 synthesis was increased at mRNA
and protein levels [5]. Immunohistochemical studies
revealed weaker staining of 5-LOX and LTA4 hydrolase in
healthy individuals and in uninvolved skin of acneic
patients than in acne lesions [5].
IL6 and IL8 are produced by SZ95 sebocytes in vitro
and their release is up-regulated in the presence of the LOX
activators AA and calcium ionophore [5]. SZ95 sebocytes
can produce cytokines without any exogenous mediation.
IL1 mRNA and protein are expressed in cultured sebo-
cytes at steady state [127]. In addition the cells were found
to release IL1, -6, -8 and TNF [5].
Neuropeptides also play an important role in induction
of inXammatory processes. Toyoda and Morohashi [100]
reviewed the involvement of neuropeptides, neuropeptides
degrading enzymes and neurotrophic factors in the inXam-
matory process of acne. Among them, substance P is
expressed in small nerves around the acne-involved seba-
ceous glands, promotes the development of cytoplasmic
organelles in sebocytes, stimulates sebaceous germinative
cells and induces signiWcant increase of sebocyte size and
sebaceous gland volume. It also increased the number of
sebum vacuoles in each diVerentiated sebaceous cell, all of
which suggest that substance P promotes both the prolifera-
tion and the diVerentiation of sebocytes. Substance P
expression has also been associated with increased enerva-
tion around the acne involved sebaceous glands. The latter
is related with increased expression of nerve growth factor
in acne prone sebaceous glands. Neutral endopeptidase, an
enzyme that degrades substance P is highly expressed in the
sebaceous glands of acne patients [101]. In addition, immu-
nohistochemical studies revealed that germinate sebocyte
cells in acne patients express nerve growth factor, which is
induced by sebocyte-produced IL6 [102].
Sebocytes through life
While the number of sebaceous glands remains the same
during life, sebum levels tend to decrease after menopause
in females, whereas no major changes appear until the
eighth decade of life in men [122]. Reduced androgen lev-
els in aged individuals lead to a slow cellular turnover in
the sebaceous glands resulting in hyperplasia of the facial
sebaceous glands in advanced age. Ultraviolet radiation and
immune suppression (cyclosporin A with corticosteroids)
Arch Dermatol Res (2008) 300:397413
represent cofactors for the development of sebaceous gland
hyperplasia. Current molecular Wndings indicate that over-
expression of the ageing-associated gene Smad7 and para-
thormone-related protein correlate with sebaceous gland
hyperplasia, whereas c-myc overexpression is associated
with enhanced sebum production.
The change of levels of IGF-I, GH, 17-estradiol, proges-
terone, DHEA and testosterone is varying in females and
males (Fig. 5). Human SZ95 sebocytes in vitro treated with
hormone levels that can be found in 60-year-old women pro-
duce less lipids than sebocytes treated with a hormone mix-
ture representing that found in serum of 20-year-old women
[57, 123]. A diVerential gene expression between SZ95 seb-
ocytes under the 20- and 60-year-old hormone mixture
detected diVerentially expressed genes, which are involved
in biological processes-hallmarks of ageing. The most sig-
niWcantly altered signalling pathway identiWed was that of
TGF-. A disturbed function of this cascade has been
associated with tumorigenesis, i.e. in pancreatic, prostate,
intestine, breast, and uterine cancer. Interestingly, genes
expressed in signalling pathways operative in age-associated
diseases such as Huntingtons disease, dentatorubral-pallid-
oluysian atrophy, and amyotrophic lateral sclerosis were
also identiWed. These data demonstrate that cultured sebo-
cytes represent an adequate model for ageing research. Hor-
mones interact in a complex fashion, and ageing may be
partly attributed to the changes in their circulating blood lev-
els. Furthermore, a disturbed hormone status may act a part
into the generation of neurodegenerative diseases. Thus, fur-
ther studies could produce a basis for an integrated and inter-
disciplinary approach to the analysis of ageing [54].
Fig. 5 Circulating hormone
levels in women and men during
life [57, 123]
407
Sebocyte culture models for drug investigation
Cell culture models, especially the internationally patented
SZ95 sebaceous gland cell line, have advanced in excellent
models to investigate new ingredients against sebostasis,
seborrhoea and acne but also against ageing skin. Labora-
tory Wndings explain the mechanisms of these disorders and
indicate the targets for activity of candidate substances
(Fig. 6). Moreover, the eVectiveness of known compounds
can be better understood (Table 4).
The most investigated drugs in vitro are retinoids.
13cRA is the most potent retinoid against acne (and sebor-
rhoea); it suppressed sebum excretion by signiWcantly
inhibiting lipogenesis and reduces sebocyte proliferation
and diVerentiation [53, 69, 124, 126]. Its exact mode of
action remained, however, until latety unknown [121]. In
vitro sebocyte research has revealed that 13cRA and atRA
inhibit the proliferation of cultured sebocytes in a dose- and
time-dependent manner [124, 126]. Marked decreases in
wax esters, a slight decrease in squalene and a relative
increase in cholesterol level have been measured. Tsukada
et al. [105] showed that 13cRA undergoes intracellular
isomerization to atRA in human sebocytes; the increased
levels of intracellular atRA act via RAR to exert its antipro-
liferative eVect on sebocytes. In addition, retinoids may
modify the diVerentiation of sebocytes in vitro by modulat-
ing keratin expression [62]. In addition, 13cRA induces a
reduction of mRNA expression of proMMP-2, proMMP-9
and -13, which are increased and possibly involved in acne
pathogenesis [71]. 13cRA causes a cell cycle arrest and
induces apoptosis in cultured sebocytes by a RAR indepen-
123
408 Arch Dermatol Res (2008) 300:397413

-MSH ACTH IL -1 , -6 , -8,

CRH TNF
-endorphin
CR H- R MC-R

E s t ro g en s -opiate R
(+) (+)

(+) 9cisRA

(+)
ER RXR

Lipids accumulation
AR atRA 13cisRA

Androgens RAR

COX LTB4, LA,

PGE2 AA, 15HETE
PPAR

Testosterone, 5- LOX / LTA4 hydrolase

D HT , c or ti s o l ,
CRH, atRA (+)
(+)

(+) (+) VPAC-R

LTB4
IGF-R
GH-R
NY-R

Insulin / IGF-I
N e u r o p ep t i d e Y GH

Fig. 6 Interaction between enzymes, membrane and nuclear receptors
as well as their ligands in human sebocytes: their inXuence on lipid
accumulation is displayed. The release of various hormones and
inXammatory mediators are also seen. R receptor, CRH corticotropin
releasing hormone, -MSH -melanocyte stimulating hormone, ACTH
adrenocotricotropic hormone, IL interleukin, TNF tumor necrosis fac-
tor-, 9cisRA 9-cis retinoic acid, atRA all-trans retinoic acid, LTB leu-
kotriene, AA arachidonic acid, LA linoleic acid, VPAC-R vasoactive
intestinal peptide receptor, GH growth hormone, NY neuropeptide Y,
IGF-I insulin like growth factor-I, PG prostaglandin, DHT 5-dihydro-
testosterone, ER estrogen receptor, AR androgen receptor, RXR reti-
noid X receptor, RAR retinoic acid receptor, PPAR peroxisome
proliferators-activated receptor, LOX lipoxygenase, LTA4 hydrolase
leukotriene A4 hydrolase, COX cyclooxygenase

dent mechanism [62]. Sebocyte apoptosis is mediated by els. They reduce lipid production and inXammatory signals,
neutrophil gelatinase-associated lipocalin [63]. when cells are grown conXuent [109].
Another family of pharmaceutical compounds been Zileuton, the only known potent PPAR ligand antago-
active on sebaceous glands are the anti-androgens which nist, was shown to inhibit LTB4 synthesis, which reduces
inhibit androgen metabolism or block the AR. Anti-andro- lipid synthesis [87, 130, 132]. On the other hand, the
gens directly inhibit the stimulatory eVect of testosterone PPAR antagonist GW0072 did not aVect sebaceous lipo-
and DHT on sebocyte proliferation [2] but lipogenesis genesis despite its ability to inhibit thiazolidinedione-
under the presence of PPAR ligands only [2]. Cyproterone induced adipocyte diVerentiation [103]. Kim et al. [46] and
acetate inhibits the activity of 3-HSD and blocks the AR RosenWeld et al. [79] tested on sebocyte-like rat preputial
[24]. The speciWc 5-reductase-1 inhibitor MK386 and the cells PPAR activators. PPAR activation was associated
dual types 1 and 2 inhibitor dutasteride succeeded to reduce with increased lipogenesis. PPAR ligands and androgens
enzyme activity in primary and SZ95 sebocytes [24, 90]. induce signiWcant lecels of diVerentiation and lipogenesis
Finasteride, a speciWc 5-reductase-2 inhibitor was able to in human sebocytes and rat cells [53, 80]. PPAR,  ligand
inhibit the enzyme activity only at very high concentration GW 2433, PPAR, ,  agonist GW 4128 and PPAR ago-
[24]. RU 58841 myristate was metabolised in cells of the nist rosiglitazone caused a signiWcant increase in the pro-
sebaceous gland cell line SZ95 to a potent antiandrogen duction of cholesterol and fatty alcohols indicating an eVect
[59]. 6-(2-adamantan-2-ylidene-hydroxybenzoxazole)-O- on the cholesterol and fatty acid biosynthetic pathway [34].
sulfamate was found to be a potent irreversible inhibitor of Additionally, rosiglitazone was able to increase signiW-
puriWed human steroid sulfatase, which is expressed in seb- cantly the triglyceride levels. Hamster sebocytes treated
ocytes, and speciWc for this enzyme relative to human aryl- with the PPAR ligand troglitazone induced signiWcantly
sulfatases A and B. In cellular assays with human sebocytes intracellular neural lipids [39]. Inhibition of the pro-inXam-
6-(2-adamantan-2-ylidene-hydroxybenzoxazole)-O-sulfa- matory and sebotropic activities of PPAR and a possible
mate blocked steroid sulfatase activity [86]. regulation of PPAR, may be more powerful than the inhi-
Corticosteroids (prednisone, prednisolone and dexa- bition of the PG pathway, including PPAR1 regulation
metasone) have also been tested in cultured sebocyte mod- [120].

123
Arch Dermatol Res (2008) 300:397413 409

Table 4 Knowledge on thera-

Compounds Used in In clinical Ruled out Still
peutic compounds occurred by
patient trials (questionable eYcacy experimental
the application of sebocyte cul-
treatment or side eVects)
ture models
Retinoids
Isotretinoin q
Antiandrogens
Androgen receptor blockers
Cyptoreone acetate q
Inhibitors of androgen metabolism
MK386 q
Finasteride q
Dutasteride qa
RU 58841 myristate q
6-(2-adamantan-2-ylidene- q
hydroxybenzoxazole)-O-sulfamate
Corticosteroids q
PPAR ligands
Zileuton qa q q
Rosiglitazone qa
Troglitazone qa
GW 2433 q
GW 4128 q
Ectopeptidase inhibitors
IP10.C8 q
Capsaicin qa q
Cannabinoids q
a
Histamine 1 receptor antagonists
Registered for other indica-
tions Diphenhydramine qa q

Thielitz et al. [96] has shown that ectopeptidease inhibi-
tors reduce proliferation and cytokine production in SZ95
sebocytes as well as the topical function of human periphe-
ral T cells in vivo and in vitro. Inhibitors of dipepditidyl
peptidase IV and aminopeptidase N are promising anti-acne
compounds.
Conclusion
This article summarizes the newest research on sebostasis,
seborrhoea and acne as well as ageing research with human
and animal sebocyte culture models from primary cells to
immortal cell lines. Interactions in sebocytes are a combi-
nation of diVerent involved pathways, signalling molecules
and genes, which may also play a role in sebaceous gland
diseases. Depending on the research target the sebocyte cul-
ture model design varies to investigate sebaceous gland
functions, such as lipogenesis, cutaneous steroidogenesis,
androgen synthesis, anti inXammatory eVects and interac-
tions with neuropeptides.
The involvement of sebocytes in stem cell research [51]
already provided explanations on the development of sebo-
cytes and the signalling pathways involved, ageing studies
on human sebocytes demonstrated of the major role of
hormone levels for gene regulation and the cellular ageing
process.
Emotional stress induces central and local expression of
CRH and other neuropeptides, which trigger inXammation.
P. acnes may produce proteins, which become active via
binding and activation of TLR. The latter stimulate the syn-
thesis of antimicrobial peptides and lipids entrenching
innate imunity.
All these current research results deliver each a piece to
the puzzle and help to understand und develop new drugs
against sebaceous gland disorders. New compounds which
are able to inhibit LTB4 synthesis, antagonize PPAR or
inhibit ectopeptidases are some new ways to treat acne.
PPAR regulation may be a pathway to modify sebaceous
lipogenesis.
In conclusion, sebocyte culture models provide new
chances for further research on sebaceous gland disorders

123
410
and acne particularly. They are useful tools for understand-
ing the pathophysiological mechanisms of these diseases.
They also facilitate the search for biologically active ingre-
dients, new pharmaceutical and cosmetic drugs for anti-
ageing, seborrheic and dry skin treatment.
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