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Original Article
J. Clin. Biochem. Nutr. | Published online: 20 June 2017 | 1–7doi: 10.3164/jcbn.1735
Advance Publication
JCBNJournal of Clinical Biochemistry and Nutrition0912-00091880-5086the Society for Free Radical Research JapanKyoto, Japanjcbn17-3510.3164/jcbn.17-35Original Article
Protective effects of astaxanthin on skin
deterioration
Kumi Tominaga,* Nobuko Hongo, Mayuko Fujishita, Yu Takahashi and Yuki Adachi
AstaReal Co., Ltd., 55 Yokohoonji, Kamiichimachi, Nakaniikawagun, Toyama 9300397, Japan
*To whom correspondence should be addressed.
Email: ktominaga@astareal.co.jp
??
(Received 18 April, 2017; Accepted 1 May, 2017)
Copyright © 2017 JCBN2017This is an open access article distributed under the terms of theCreative Commons Attribution License, which permits unre-stricted use, distribution, and reproduction in any medium, pro-vided the original work is properly cited.
Astaxanthin is a carotenoid with potent antioxidant and anti
inflammatory activity. To evaluate the antiinflammatory effect
of astaxanthin on skin deterioration, we confirmed its role in
epidermaldermal interactions in vitro. Astaxanthin treatment
suppressed ultraviolet B (UVB)induced inflammatory cytokine
secretion in keratinocytes, and matrix metalloproteinase1 secre
tion by fibroblasts cultured in UVBirradiated keratinocyte medium
.
To verify these findings, we conducted a 16week clinical study
with 65 healthy female participants. Participants were orally
administered either a 6 mg or 12 mg dose of astaxanthin or a
placebo. Wrinkle parameters and skin moisture content signifi
cantly worsened in the placebo group after 16 weeks. However,
significant changes did not occur in the astaxanthin groups.
Interleukin1
α
levels in the stratum corneum significantly increased
in the placebo and lowdose groups but not in the highdose group
between weeks 0 and 16. This study was performed in Japan from
August to December, when changing environmental factors, such
as UV and dryness, exacerbate skin deterioration. In conclusion,
our study suggests that longterm prophylactic astaxanthin
supplementation may inhibit agerelated skin deterioration and
maintain skin conditions associated with environmentally induced
damage via its antiinflammatory effect. (UMIN Clinical Trials
Registry ID: UMIN000018550)
Key Words: astaxanthin, inflammatory cytokines,
wrinkle formation, skin elasticity, interleukin1α
Introduction
A
staxanthin is a naturally occurring xanthophyll carotenoid
that was originally isolated from lobsters by Kuhn and
Sorensen.
(1)
It is found in crustaceans, such as shrimps and crabs,
and fish, such as salmon and sea bream. The anti-inflammatory
activity of astaxanthin, which is based on its antioxidant pro-
perties, has been implicated in improving lifestyle-related diseases
and managing health. Furthermore, astaxanthin has anti-aging
effects.
(2)
Mechanisms of skin aging are classified into physiological and
photoaging.
(3–5)
Skin aging manifests as wrinkles, degradation of
elasticity, and age spots (liver spots). Physiological skin aging
occurs over the whole body and is a result of decreased cellular
metabolism. Photoaging occurs in ultraviolet (UV)-irradiated
areas of the skin, such as the face. It is primarily due to collagen
and elastic fiber degradation that occurs because of matrix metal-
loproteinases (MMPs) secreted from dermal fibroblasts and
epidermal keratinocytes in response to UV irradiation.
(6,7)
In addi-
tion, MMP secretion and activation are stimulated by the various
inflammatory cytokines secreted from keratinocytes by reactive
oxygen species (ROS) from UV-irradiated cells.
(8)
Skin inflam-
mation is triggered by oxidative stress that results from stimuli
such as UV exposure and skin dryness. Thus, suppression of
inflammatory cytokines by oxidative stress inhibition is crucial for
inhibiting age-related skin deterioration.
The effects of astaxanthin on hyper-pigmentation suppression,
melanin synthesis and photoaging inhibition, and wrinkle forma-
tion reduction have been reported in several clinical studies.
(9,10)
In
this study, we conducted in vitro and in vivo studies to investigate
the effects of astaxanthin on skin deterioration. We assessed
the impact of astaxanthin on inflammatory cytokine and MMP-1
expression in UVB-irradiated human keratinocytes and human
dermal fibroblasts, respectively. Furthermore, we conducted a
randomized, double-blind, parallel-group, placebo-controlled study
involving 65 healthy female participants for 16 weeks to investigate
the in vivo effect of oral astaxanthin supplementation.
Materials and Methods
Chemicals and materials.
Minimum essential medium alpha
(MEMα) and Dulbecco’s modified Eagle’s medium (DMEM)
were purchased from Life Technologies (Grand Island, NY), and
fetal bovine serum (FBS) was obtained from Nichirei Biosciences
(Tokyo, Japan). Human epidermal keratinocytes and KG2 medium
were obtained from Kurabo (Osaka, Japan). Human dermal fibro-
blasts were obtained from the Riken BRC (Tsukuba, Japan).
Dimethyl sulfoxide (DMSO) was purchased from Wako Pure
Chemical Industries (Osaka, Japan). The enzyme-linked immuno-
sorbent assay (ELISA) kits for interleukin (IL)-1α, IL-6, IL-8,
and tumor necrosis factor (TNF)-α were purchased from R&D
Systems (Minneapolis, MN). The MMP-1 ELISA kit was purchased
from GE Healthcare (Buckinghamshire, England). Mammalian
protein extraction reagent (M-PER) and the BCA protein assay
kit were purchased from Thermo Scientific Pierce (Rockford,
IL). Astaxanthin and all other reagents were purchased from
Sigma-Aldrich (St. Louis, MO).
Keratinocyte culture and UVB irradiation. Keratinocytes
were cultured in KG2 medium according to the manufacturer’s
protocols and incubated at 37°C and 5% CO
2
. Cells were plated
on 60-mm dishes at a density of 3.5 ×10
5
cells/dish and incubated
for 48 h before UVB treatment. Fresh KG2 medium containing
various concentrations of astaxanthin (0, 1, 5 and 10 μM) and
0.5% (v/v) DMSO as a vehicle was then added to keratinocytes,
and cells were incubated for another 4 h. Keratinocytes were
washed once with phosphate-buffered saline (PBS), 500 μl PBS
was added, and the cells were then irradiated with 5 mJ/cm
2
UVB. PBS was immediately replaced with fresh KG2 medium
containing various concentrations of astaxanthin (0, 1, 5 and
10 μM) and 0.5% DMSO, and keratinocytes were cultured for
24 h. The culture medium was harvested and frozen at –20°C until
subsequent analysis. Keratinocytes were rinsed once with PBS
and lysed in 1.0 ml M-PER.
A
doi: 10.3164/jcbn.17352
UVB source. Keratinocyte irradiation was performed using
two UVB lamps (TL20W/12RS; Philips Lighting, Holding,
Amsterdam, Netherlands). UVB irradiance was measured using
the UV light meter UV-340 (Lutron Electronics, Coopersburg, PA).
Indirect treatment of fibroblasts with UVBirradiated
keratinocytes. Fibroblasts were cultured in MEMα supple-
mented with 10% FBS at 37°C and 5% CO
2
. Cells were grown in
48-well plates at a density of 2.4 ×10
3
cells/well and cultured
overnight before treatment. To investigate the indirect effect of
astaxanthin on MMP-1 production by fibroblasts as mediated by
keratinocytes, keratinocytes were pre-treated with astaxanthin
and irradiated with UVB, as described above. Following UVB
irradiation, keratinocytes were cultured for 24 h in fresh DMEM
containing various concentrations of astaxanthin and 0.5%
DMSO. The keratinocyte culture medium was collected and
immediately transferred to fibroblasts. After 48 h, the fibroblast
culture medium was harvested and frozen at –20°C until the
MMP-1 assay. Fibroblasts were rinsed once with PBS and lysed in
1.0 ml M-PER.
ELISA and protein assay. Levels of IL-1α, IL-6, IL-8 and
TNF-α secreted from keratinocytes were measured by ELISA
as per the manufacturer’s instructions. MMP-1 levels secreted
from fibroblasts were also measured by ELISA according to the
manufacturer’s instructions. ELISA data were normalized to the
total protein content in the cell lysate corresponding to each
sample medium. Protein concentrations were measured using the
BCA protein assay kit.
Clinical study of astaxanthin supplementation. A ran-
domized, double-blind, parallel-group, placebo-controlled study
was conducted to evaluate the effects of astaxanthin on wrinkle
formation and other aspects of skin damage and aging. Before
starting the clinical test, the wrinkle grades on the lower and outer
angle eyelids of subject candidates were evaluated by a trained
expert. Sixty-five healthy female participants (age, 35–60 years)
with a wrinkle grade of 2.5 to 5.0 were enrolled. Written informed
consent was obtained, and the study was approved by the Institu-
tional Review Board of the Tokyo Synergy Clinic, Tokyo, Japan.
Measurements were performed at three key points in the study:
before treatment (week 0), after 8 weeks (week 8), and after 16
weeks (week 16). A biochemical examination of blood serum
chemistry was also performed to assess astaxanthin supplementa-
tion safety at weeks 0 and 16. The study period was from August
to December 2015 at the research institution in Osaka, Japan.
Material for oral supplementation. The material for oral
supplementation contained AstaReal
®
Oil 50F (Fuji Chemical
Industries, Toyama, Japan), 5% w/w astaxanthin Haematococcus
pluvialis extract, and canola oil as soft gel capsules. Participants
were randomly assigned to one of three groups: high-dose
(n= 22), low-dose (n= 22), or placebo (n= 21), in which partici-
pants were administered daily oral supplements containing 12, 6
or 0 mg astaxanthin, respectively, for 16 weeks.
Wrinkle analysis and instrumental measurements. For
screening of subjects for the wrinkle grade, evaluators used the
wrinkle grade standard proposed by the Japanese Cosmetic
Science Society
(11)
using a photo of each participant that was
taken using VISIA
®
Evolution (Canfield Scientific, NJ). During
the study period, the following measurements were performed:
replicas of wrinkles of the same area were obtained, and four
parameters (area ratio of all wrinkles, mean depth of the deepest
wrinkle, maximum depth of the deepest wrinkle, and mean depth
of all wrinkles) were calculated by image analysis using PRIMOS
LITE (GFMesstechnik, Teltow, Germany). Capacitance of the
cheek, which indicates skin moisture content, was measured using
a skin hygrometer SKICON-200EX (YAYOI, Tokyo, Japan).
Transepidermal water loss at the cheek was measured using AS-
CT1 (Asahibiomed, Tokyo, Japan). Skin elasticity was measured
using a Cutometer MPA560 (Courage+Khazaka electronic,
Cologne, Germany), and three parameters, R2, R6 and R7, which
represent gross elasticity, portion of the viscoelasticity, and
biological elasticity, respectively, were calculated. The stratum
corneum of each subject was obtained using the adhesive Skin
Checker (Promotool Corporation, Tokyo, Japan), and IL-1α
levels in the stratum corneum were measured by ELISA according
to the manufacturer’s instructions.
Statistical analyses. All data are reported as mean ±SD.
Statistical tests were performed using Dunnett’s test or Student’s
t test, and a p value of <0.05 was considered to be statistically
significant.
Results
Effects of astaxanthin supplementation on inflammatory
cytokine production in UVBirradiated keratinocytes.
Keratinocytes were treated for 4 h with 0, 1, 5 or 10
µ
M astaxanthin
and then irradiated with 5 mJ/cm
2
UVB. The culture medium was
collected, and the levels of inflammatory cytokines, including
IL-1α, IL-6, IL-8 and TNF-α, were measured. All cytokine levels
in the culture medium significantly increased following UVB irra-
diation compared with those in the non-irradiated medium. IL-1α,
IL-6, IL-8 and TNF-α levels significantly decreased following
treatment with astaxanthin in a dose-dependent manner (Fig. 1).
Effects of astaxanthin supplementation on fibroblast
MMP1 production. Fibroblasts were incubated with media
derived from keratinocytes that were treated with 0, 1, 5 or 10 μM
astaxanthin before and after irradiation with UVB. The fibroblast
culture medium was then collected and assessed for MMP-1 pro-
duction. MMP-1 levels significantly increased in the medium
derived from UVB-irradiated keratinocytes. However, MMP-1
levels significantly decreased in a dose-dependent manner in the
presence of medium derived from astaxanthin-treated keratino-
cytes (Fig. 2).
Clinical efficacy of oral astaxanthin supplementation on
facial skin. Of 66 individuals who volunteered for the study,
59 participants were included in our clinical study (Fig. 3). The
wrinkle grades were 3.45 ±0.71 in the placebo group, 3.43 ±0.70
in the low-dose group, and 3.38 ±0.68 in the high-dose group
at week 0. Mean and maximum depths of the deepest wrinkle
significantly deteriorated in the placebo group at week 16 com-
pared with those at week 0 (Fig. 4). Furthermore, skin moisture
content displayed significant deterioration in the placebo group
at week 16 (204.9 ±54.2 μS) compared with that at week 0
(264.7 ±100.3 μS). In contrast, significant deteriorations in
wrinkle parameters from replica image analysis and skin moisture
content (data not shown) were not observed in the astaxanthin-
treated groups, indicating that astaxanthin maintained skin
conditions during the study period. Skin elasticity was evaluated
using three parameters, R2, R6 and R7. R2 and R6 values signifi-
cantly improved in all groups at week 8, but R6 and R7 values
significantly improved at week 16 compared with those at week 0
only in the low-dose group (Fig. 5). In addition, IL-1α levels in
the stratum corneum significantly deteriorated in the placebo and
low-dose groups at week 16, but did not change in the high-dose
group during the study period (Fig. 6). There were no changes in
transepidermal water loss in any group (data not shown). No
measured parameters, including wrinkle parameters, skin moisture
content, transepidermal water loss, skin elasticity, and IL-1α
levels, displayed significant differences when comparing among
the groups at weeks 0, 8 and 16.
Stratified analyses. The stratified analysis was conducted
for participants whose capacitance at week 0 was higher than
the mean capacitance of all subjects (264.8 μS). There were 10
participants in the placebo group, 7 in the low-dose group and 11
in the high-dose group. Mean depth of the deepest wrinkle,
maximum depth of the deepest wrinkle and mean depth of all
wrinkles significantly worsened in the placebo group at week 16
(68.6 ±23.0, 172.0 ±53.6 and 59.9 ±19.8 μm, respectively) com-
J. Clin. Biochem. Nutr. | Published online: 20 June 2017 | 3K. Tominaga et al.
pared with those at week 0 (43.6 ±11.6, 108.6 ±29.2 and 42.9 ±
13.7
µ
m, respectively). In contrast, significant worsening of wrinkle
parameters was not observed in the astaxanthin-treated group
(data not shown). R2 and R6 values significantly improved only in
the low-dose group at week 16, and the R7 value deteriorated only
in the placebo group at week 8 compared with those at week 0
(Fig. 7). Moreover, the R2 value displayed a significant improve-
ment in the high-dose group (p= 0.001) at week 8 compared
with that in the placebo group. In addition, the R7 value displayed
a significant improvement in the high-dose group at week 8
(p= 0.002) and a tendency to improve in the low-dose group at
week 16 (p= 0.100) compared with that in the placebo group.
Safety evaluation of astaxanthin supplementation.
Results of the biochemical examination of blood are summarized
in Table 1. There were no abnormal changes in blood parameters
observed during the study period, and no serious adverse events
were reported.
Discussion
We conducted in vitro and in vivo studies to investigate the
effects of astaxanthin, a natural antioxidant, on skin deterioration
observed in aged skin. We demonstrated that pre- and post-
treatment with astaxanthin dose-dependently suppressed the
secretion of inflammatory cytokines from UVB-irradiated keratino-
cytes
. Furthermore, MMP-1 production by fibroblasts that were
treated with medium from UVB-irradiated keratinocytes with
astaxanthin treatment decreased in a dose-dependent manner.
Inflammatory cytokines released from epidermal keratinocytes
stimulate dermal fibroblasts and keratinocytes in an autocrine
manner and then upregulate messenger ribonucleic acid expres-
sion, protein, and enzymatic activity levels of MMPs, such as
MMP-1, MMP-3 and MMP-9. MMPs subsequently affect collagen
and elastic fibers, leading to the formation of wrinkles.
(12)
UV-
Fig. 1. Effect of astaxanthin on cytokine production in UVBirradiated keratinocytes. Each value represents mean value ±SD, **p<0.01 and
*p<0.05 by ANOVA/Dunnett’s test. ANOVA, analysis of variance; AX, astaxanthin; IL, interleukin; TNF, tumor necrosis factor; UV, ultraviolet.
Fig. 2. Effect of conditioned media from UVBirradiated keratinocytes
treated with or without astaxanthin on MMP1 production in cultured
fibroblasts. Each value represents mean value ±SD, *p<0.01 by ANOVA/
Dunnett’s test. ANOVA, analysis of variance; AX, astaxanthin; MMP,
matrix metalloproteinase; UV, ultraviolet.
doi: 10.3164/jcbn.17354
induced wrinkle formation is markedly inhibited by elastase
activity suppression in degrading elastic fibers. Previous study
indicated that qualitative and quantitative changes in elastic fibers
caused wrinkle formation. Our in vitro study indicated that UVB
irradiation-induced inflammation and inflammatory cytokine-
stimulated MMP-1 levels were suppressed by astaxanthin. These
results corroborated those of previous studies.
(13–16)
In addition, it
was reported that long-term treatment with astaxanthin prevented
the accumulation of age-related oxidative stress and inflammatory
response in aging mice.
(17)
Thus, based on these collective results,
oral astaxanthin supplements are expected to inhibit inflam-
mation-mediated skin deterioration, such as wrinkle formation and
skin moisture decline, that appears in aged skin.
Next, we conducted a randomized, double-blind, parallel-group
,
Fig. 3. Flow diagram of the clinical trial.
Fig. 4. Effect of oral astaxanthin supplementation on wrinkle parameters from replica image analysis. Each value represents mean value ±SD,
*p<0.05 by ANOVA/Dunnett’s test. ANOVA, analysis of variance.
J. Clin. Biochem. Nutr. | Published online: 20 June 2017 | 5K. Tominaga et al.
placebo-controlled study with 65 healthy female subjects for 16
weeks to verify the effects of oral astaxanthin supplementation on
skin integrity. We determined that skin moisture content
and
deep wrinkles were not significantly changed in the astaxanthin-
supplemented groups, whereas these parameters significantly
worsened in the placebo group during the study period. Further-
more, IL-1α levels in the stratum corneum were maintained only
in the high-dose group. In addition, skin elasticity improvements
were observed in high-dose group compared with that of the
placebo group in participants with high skin moisture content.
It is well known that both UV radiation and dryness cause
progression of wrinkle formation. UV irradiation contributes to
wrinkle formation by inducing MMP secretion from dermal fibro-
blasts via cytokines, such as IL-1α, IL-6, and TNF-α, released by
UVB-exposed keratinocytes.
(4)
In the region where this study was
conducted, daytime UV light is the strongest between May and
September, and the air humidity declines from August, reaching
the lowest levels between December and April. Our clinical trial
was conducted from August to December, a period during which
exposure to strong UV radiation during the summer is followed
by a decrease in humidity during the autumn and winter months.
As a result of these changing environmental factors, skin barrier
function declines and skin dryness progresses.
(18)
Exposure to low
humidity induces epidermal IL-1α synthesis, and water flux in
the epidermis might be the first signal to induce IL-1α synthesis in
the epidermis.
(19)
IL-1α may also induce other proinflammatory
cytokines, such as IL-6 and IL-8.
(20)
Thus, the IL-1α level in the
stratum corneum is linked to skin dryness, which we also observed
Fig. 5. Effect of oral astaxanthin supplementation on skin elasticity. Each value represents mean value ±SD, **p<0.01 and *p<0.05 by ANOVA/
Dunnett’s test. ANOVA, analysis of variance.
Fig. 6. Effect of oral astaxanthin supplementation on IL1α in the
stratum corneum measured by ELISA. Each value represents mean
value ±SD, **p<0.01 and *p<0.05 by paired t test. ELISA, enzymelinked
immunosorbent assay; IL, interleukin.
Fig. 7. Stratified analysis of skin elasticity parameters of the cheek. Each value represents mean value ±SD, **p<0.01 and *p<0.05 by ANOVA/
Dunnett’s test. R2, R6 and R7 represent gross elasticity, portion of the viscoelasticity, and biological elasticity, respectively. ANOVA, analysis of
variance.
doi: 10.3164/jcbn.17356
Table 1. Summary of the biochemical blood exam
AST, aspartate aminotransferase; ALT, alanine aminotransferase; ALP, alkaline phosphatase; LD, lactate dehydrogenase; γGT, gummaglutamyl
transferase; CK, creatine kinase; A/G, albumin/globulin ratio; ALB, albumin; HbA1c, hemoglobin A1c; HDL, high density lipoprotein; LDL, low density
lipoprotein; Na, sodium ; K, potassium; Cl, chlorine; Ca, calcium.
Group n
Measured value Comparison between
week 0 and 16 Intergroup comparison
of change amount
Week 0 Week 16
Mean SD Mean SD p value p value
Total bilirubin (mg/dl) Placebo 18 0.76 ±0.24 0.67 ±0.15 0.064
Lowdose 22 0.75 ±0.22 0.74 ±0.21 0.696 0.370
Highdose 19 0.71 ±0.20 0.71 ±0.16 1.000 0.791
AST (U/L) Placebo 18 19.89 ±3.48 21.56 ±5.19 0.053
Lowdose 22 20.82 ±7.22 21.32 ±4.22 0.678 0.979
Highdose 19 18.21 ±4.81 20.00 ±3.80 0.008*0.457
ALT (U/L) Placebo 18 14.11 ±4.54 15.78 ±7.95 0.219
Lowdose 22 17.64 ±11.56 15.77 ±7.12 0.247 1.000
Highdose 19 13.95 ±4.96 14.79 ±4.22 0.353 0.86
ALP (U/L) Placebo 18 185.28 ±60.11 191.06 ±59.17 0.234
Lowdose 22 191.68 ±71.31 185.32 ±59.43 0.325 0.923
Highdose 19 179.58 ±38.65 193.26 ±43.91 0.001*0.989
LD (LDH) (U/L) Placebo 18 166.83 ±16.04 161.72 ±20.36 0.149
Lowdose 22 181.82 ±22.56 173.95 ±25.23 0.016*0.291
Highdose 19 165.00 ±31.11 166.47 ±36.13 0.777 0.823
γGT (U/L) Placebo 18 24.17 ±24.06 23.22 ±21.29 0.365
Lowdose 22 27.09 ±27.56 22.41 ±15.60 0.151 0.979
Highdose 19 16.00 ±3.54 15.53 ±4.05 0.283 0.224
CK (U/L) Placebo 18 92.11 ±46.07 96.39 ±58.94 0.438
Lowdose 22 92.27 ±44.62 100.86 ±46.12 0.318 0.933
Highdose 19 96.16 ±39.00 89.53 ±29.70 0.275 0.861
Total protein (g/dl) Placebo 18 7.40 ±0.38 7.50 ±0.41 0.252
Lowdose 22 7.50 ±0.32 7.57 ±0.38 0.296 0.774
Highdose 19 7.39 ±0.43 7.66 ±0.36 0.012*0.356
A/G Placebo 18 1.45 ±0.18 1.41 ±0.20 0.304
Lowdose 22 1.41 ±0.11 1.43 ±0.18 0.718 0.967
Highdose 19 1.43 ±0.19 1.41 ±0.21 0.453 0.997
ALB (g/dl) Placebo 18 4.36 ±0.22 4.37 ±0.20 0.895
Lowdose 22 4.39 ±0.21 4.43 ±0.18 0.346 0.467
Highdose 19 4.33 ±0.21 4.45 ±0.21 0.010*0.305
Creatinine (mg/dl) Placebo 18 0.69 ±0.10 0.64 ±0.08 0.001*
Lowdose 22 0.67 ±0.08 0.64 ±0.07 0.011*0.999
Highdose 19 0.69 ±0.08 0.65 ±0.10 0.003*0.995
Urea nitrogen (mg/dl) Placebo 18 13.28 ±3.01 12.56 ±2.66 0.200
Lowdose 22 13.27 ±2.39 12.73 ±2.90 0.352 0.970
Highdose 19 13.00 ±2.60 12.11 ±2.42 0.145 0.826
Uric acid (mg/dl) Placebo 18 4.36 ±1.05 4.22 ±1.11 0.269
Lowdose 22 4.60 ±0.73 4.45 ±0.92 0.071 0.701
Highdose 19 4.23 ±0.92 4.28 ±0.97 0.506 0.974
Fasting glucose (mg/dl) Placebo 18 88.06 ±7.69 87.89 ±9.50 0.886
Lowdose 22 87.18 ±8.54 87.23 ±9.73 0.975 0.967
Highdose 19 87.26 ±7.27 90.05 ±10.38 0.206 0.726
HbA1c (%) Placebo 18 5.37 ±0.28 5.37 ±0.32 1.000
Lowdose 22 5.34 ±0.20 5.32 ±0.25 0.411 0.794
Highdose 19 5.31 ±0.23 5.27 ±0.25 0.187 0.474
Total cholesterol (mg/dl) Placebo 18 234.22 ±50.79 231.50 ±49.64 0.286
Lowdose 22 230.41 ±32.68 228.82 ±28.91 0.590 0.960
Highdose 19 211.37 ±24.99 218.53 ±28.03 0.059 0.446
Triglyceride (mg/dl) Placebo 18 76.44 ±27.65 72.56 ±30.31 0.428
Lowdose 22 71.00 ±24.80 71.14 ±30.84 0.977 0.983
Highdose 19 68.84 ±24.26 64.26 ±28.39 0.399 0.606
HDL cholesterol (mg/dl) Placebo 18 72.78 ±11.40 76.22 ±13.32 0.011*
Lowdose 22 75.23 ±10.77 80.41 ±12.40 0.008*0.540
Highdose 19 73.63 ±15.84 83.00 ±16.21 0.000*0.247
LDL cholesterol (mg/dl) Placebo 18 140.89 ±47.68 141.11 ±47.39 0.939
Lowdose 22 136.77 ±30.67 136.32 ±30.56 0.830 0.872
Highdose 19 119.79 ±21.30 122.53 ±24.68 0.419 0.193
Arteriosclerosis index Placebo 18 2.00 ±0.76 1.93 ±0.80 0.159
Lowdose 22 1.87 ±0.54 1.76 ±0.57 0.059 0.642
Highdose 19 1.73 ±0.62 1.55 ±0.58 0.005*0.144
Na (mEq/L) Placebo 18 140.61 ±1.33 139.17 ±1.72 0.000*
Lowdose 22 140.32 ±1.62 140.55 ±1.26 0.424 0.019*
Highdose 19 140.11 ±1.59 140.00 ±1.91 0.772 0.217
K (mEq/L) Placebo 18 4.12 ±0.26 4.18 ±0.27 0.373
Lowdose 22 4.24 ±0.32 4.32 ±0.27 0.178 0.222
Highdose 19 4.17 ±0.19 4.26 ±0.32 0.258 0.567
Cl (mEq/L) Placebo 18 102.39 ±1.72 102.28 ±1.74 0.742
Lowdose 22 102.86 ±1.55 103.23 ±1.85 0.257 0.198
Highdose 19 102.53 ±1.58 102.32 ±2.00 0.600 0.997
Ca (mEq/L) Placebo 18 9.58 ±0.28 9.39 ±0.23 0.004*
Lowdose 22 9.74 ±0.35 9.50 ±0.26 0.000*0.392
Highdose 19 9.67 ±0.38 9.62 ±0.34 0.434 0.034*
Serum iron (μg/dl) Placebo 18 122.61 ±45.95 94.22 ±35.22 0.009*
Lowdose 22 106.05 ±40.02 118.32 ±46.81 0.226 0.126
Highdose 19 105.11 ±39.92 102.58 ±39.65 0.752 0.760
J. Clin. Biochem. Nutr. | Published online: 20 June 2017 | 7K. Tominaga et al.
in the placebo and low-dose groups, even though wrinkle para-
meters in the astaxanthin-treatment groups were not significantly
altered.
With respect to safety, no adverse events were observed with
oral astaxanthin supplementation of 12 mg/day for 16 weeks. Our
results confirm the long-term safety of astaxanthin as an oral
supplement.
In conclusion, our findings indicated that astaxanthin inhibited
inflammatory cytokine secretion from epidermal keratinocytes
and MMP-1 secretion by dermal fibroblasts in response to UVB
irradiation. These mechanisms underlie clinical study results that
demonstrated that the suppression of seasonal deterioration of
wrinkles and skin moisture and the improvements in skin elasticity
were accompanied by sustaining low IL-1α levels in the epidermal
corneum. Thus, long-term astaxanthin supplementation may
prophylactically inhibit skin deterioration induced over time by
environmental damage and consequently retard the skin aging
process via its anti-inflammatory effect.
Abbreviations
BCA bicinchoninic acid
DMEM Dulbecco’s modified Eagle’s medium
DMSO dimethyl sulfoxide
ELISA enzyme-linked immunosorbent assay
FBS fetal bovine serum
IL interleukin
MEMαminimum essential medium alpha
MMP matrix metalloproteinase
M-PER mammalian protein extraction reagent
PBS phosphate-buffered saline
ROS reactive oxygen species
TNF tumor necrosis factor
UV ultraviolet
Conflict of Interest
This study was financially sponsored by Fuji Chemical Industries
Co., Ltd.
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