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A Hair Growth-Promoting Effect of Chinese Black Tea Extract in Mice
I-Ching HOU,
1;2;
y
Yasuyuki OI ,
1
Hiroyuki FUJITA,
1
Yoshihisa YANO,
2
Harukazu FUKAMI,
2
and Masaaki YOSHIKAWA
3
1
Research and Development Department, Nippon Supplement, Inc.,
Applause Tower 18 F, 19-19, Chayamachi, Kita-ku, Osaka 530-0013, Japan
2
Graduate School of Bioenvironmental Science, Kyoto Gakuen University,
1-1 Nanjyo-otani, Sogabe-cho, Kameoka, Kyoto 621-8555, Japan
3
Division of Functional Food Science, Research Institute for Production Development,
15 Shimogamo-morimoto-cho, Sakyo-ku, Kyoto 606-08 05, Japan
Received April 12, 2013; Accepted June 10, 2013; Online Publication, July 7, 2013
[doi:10.1271/bbb.130298]
Chinese black tea extract (CBTE) fermented with
Aspergillus sp. significantly promoted hair growth after 2
weeks of topical application in shaved 6 week-old male
C3H/He mice. The hair growth-promoting effect of
CBTE was potentiated synergistically by capsaicin, which
has no effect on hair growth by itself. CBTE displayed an
affinity for estrogen receptor (ER), with an IC
50
value
of 74.8 g/mL. This effect of CBTE might be mediated by
the ERs, since a similar effect induced by orally admin-
istered soy isoflavone, a mixture of ERs ligands, has been
reported to be synergistically potentiated by capsaicin.
Key words: hair growth; Chinese black tea extract
(CBTE); estrogen receptor; C3H/He mice;
capsaicin
Alopecia is a common problem. Although various hair
growth-promoting substances have been reported,
1–3)
we
sought an improved approach for people who continue
to have a problem with hair growth.
It is known that flavonoids, such as the green tea
ingredient ()-epigallocathechin gallate (EGCG), pro-
mote hair growth after oral administration
4,5)
but the hair
growth-promoting effect of teas by topical application
has rarely been reported, although this administration
route is helpful in limiting the hair growth to the desired
region. In this study, we investigate the effect of an
extract of the well known Chinese black tea, Pu-erh tea,
a traditional tea fermented with Aspergillus sp., on hair
growth in shaved male C3H/He mice, administered
topically.
It has been reported that capsaicin augments the hair
growth-promoting effect of soy isoflavone, an ER
agonist, by synergistically potentiating insulin-like
growth factor I (IGF-1) release via stim ulation of
calcitonin gene- related peptide (CGRP) release from
sensory nerve endi ngs, without showing any effect on
hair growth by itself.
6)
Hence we tested the effect of
capsaicin on the hair growth-promoting effect of CBTE
to gain insight into its mode of action.
CBTE was prepared as described previously.
7,8)
CBTE powder was mixed into a basal solvent containing
propylene glycol, EtOH, and distilled water at a ratio of
5:3:2, and this was centrifuged for 10 min at 9;800 g
to give 20% (w/w). The supernatant was used to prepare
the samples. Six-week old C3H/He male mice in
the telogen phase were purchased from Japan SLC
(Shizuoka, Japan). The mice were housed in plastic
cages (four mice per cage) in a room with controlled
temperature, air flow, and humidity under a 12-h light/
12-h dark cycle, and were fed ad libitum with MF pellets
(Oriental Yeast, Osaka, Japan). The care and handing of
the animals were in accordance with Notification no. 88
of the Ministry of Environment of Japan (2006) and the
Guidelines for the Proper Conduct of Animals Experi-
ments of Kyoto Gakuen University. Three days after
shaving, 150
mL of the sample solution was applied
topically to the backs of the mice daily for 2 weeks.
Changes in the dorsa l area of the mice were observed
and recorded by visual inspection and photography.
The ratio of the hair growth area was calculated by a
comparison of the hair growth and shaved areas, which
were measured by means of image analysis software.
Data were expressed as mean SD. Student’s t test was
used to determine statistical significance.
As shown in Fig. 1, the application of 3.5% CBTE
cause a significant increase after 2 weeks of topical
application (p < 0:05). As reported for the hair growth-
promoting effect of soy isoflavone, 0.05% capsaicin
synergistically potentiated CBTE, although the effect
was not significant. Even though an inhibitory effect
of capsaicin on hair-shaft elongation of human hair
follicles in vitro has been reported,
9)
no such effect was
observed in our in vivo system.
Next we investigated the affinities of CBTE for the
ER receptor, since its hair growth-pr omoting effect, as
well as that of soy isoflavone, acting as an ER agonist,
was potentiated by capsaicin, as described above. The
affinity of CBTE for the ER receptor was evaluated by
radioligand binding assays using human recombinant
ER receptor expressed in insect Sf9 cel ls, along with
0.5 n
M [
3
H]-estradiol. Non-specific binding was estimated
in the presence of 1 mM diethylstilbestrol, as previously
described.
10)
The IC
50
value was determined by non-
y
To whom correspondence should be addressed. Fax: +81-6-6376-2234; E-mail: hou@nippon-sapuri.co.jp
Abbreviations: CBTE, Chinese black tea extract; EGCG, ()-epigallocathechin gallate; ECG, epicatechin gallate; IGF-1, insulin-like growth factor
I; CGRP, calcitonin gene-related peptide
Biosci. Biotechnol. Biochem., 77 (7), 1606–1607, 2013
Communication
linear, least squares regression analysis using MathIQ
(ID Business Solutions, Guildford, UK).
CBTE displayed an affinity for ER,withanIC
50
value of 74.8 mg/mL, irrespective of whether it is
agonistic or anta gonistic. ERs have been reported to be
related to hair growth.
11–14)
ER ligands, such as
isoflavone, have been found to exert hair growth-
promoting effects in mice.
6)
These results suggest that
the hair growth-promoting effect of CBTE might also be
mediated by ERs. EGCG and epicatechin gallate (ECG)
have been reported to have affinities for ER,withIC
50
values of 480 mM and 550 mM respectively,
15)
but these
catechins were not detected in CBTE by high perform-
ance liquid chromatography (data not shown). There-
fore, active substances showing an affinity for the ERs
might be produced during the course of fermentation in
the course of CBTE production.
Isolation of the constituents in CBTE that are
essential for its hair growth-promoting and ER binding
activities is currently underway in our laboratory.
Investigation of the details of the mechanism of the
hair growth-promoting effect of CBTE awaits isolation
of the active substances.
Acknowledgments
Financial support was provided by Nippon Supple-
ment, Inc. There was no conflict of interest.
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B
A
Fig. 1. Effects of CBTE and Capsaicin on Hair Growth.
Dorsal hair of 6-week old C3H/He mice, approximately 25 g in
body weight, was removed using an electric clipper and shavers after
anesthetization by intraperitoneal injection of thiopental sodium
(60 mg/kg BW). The mice were treated with a topical application of
3.5% CBTE and 0.05% capsaicin for 14 d (150
mL), and the skin on
the back was then photographed (A). The ratio of the hair growth
area of mice (B) was calculated by the following formula: haired
area shaved area. Data were expressed as the mean SD (n ¼ 8).
p < 0:05 vs. control.
p < 0:01 vs. control.
Hair Growth Promotion by Chinese Black Tea 1607
