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Pharmacology & Pharmacy, 2018, 9, 428-439
http://www.scirp.org/journal/pp
ISSN Online: 2157-9431
ISSN Print: 2157-9423
DOI:
10.4236/pp.2018.910032 Oct. 26, 2018 428
Pharmacology & Pharmacy
Anti-Aging Effect of Cynara cardunculus L. var.
Cynara scolymus L. Extract in
D-Galactose-Induced Skin Aging Model in Rats
Galina Sukoyan1*, Edisher Tsivtsivadze1, Veronika Golovach1, Tamara Kezeli2, Natalya Demina3
1International Center of Introduction of New Biomedical Technology, Tbilisi, Georgia
2Department of Pharmacology, Ivane Javakhishvili Tbilisi State University, Tbilisi, Georgia
3Department of Pharmacy Technology of Federal State Autonomous Educational Institution of Higher Education, I. M. Sechenov
of the Ministry of Health of the Russian Federation (Sechenov University), Moscow, Russia
Abstract
The aim of the study was to evaluate the efficacy of 2%
Cynara scolymus
L.
extracts in reverse disturbances of collagen metabolism and inflammation in
D-galactose induced skin aging model in rats. D-galactose induced aging re-
produced in 48 animals of main group, and 12 rats in control group. All ani-
mals in main group were randomized for 4 groups: I. aging + saline, II - IV.
different manufacturers 2% artichoke extracts (with content of
chlorogenic
acid < 2.5%) in a dose of intradermal injection 0.13 mg twice at weeks during
4 weeks. Influence of artichoke extracts restored skin relative weight and led
to an increase of solubility in neutral salt and acid, and decreased pepsin so-
lubility collagen fraction, restored the hexosamine/collagen (hydroxyproline)
ratio and decreased the activity of nuclear transcription factor (NF-
kB). Local
prolonged treatment with artichoke ex
tracts improved collagen metabolism
and attenuated the progression of inflammation in D-
galactose induced skin
aging model.
Keywords
Skin, Aging, Collagen, Pure Water
Cynara cardunculus
L. var.
Cynara
scolymus
L. Extract, Nuclear Transcription Factor Kappa B
1. Introduction
Intrinsic or innate aging, an unpreventable process, affects the skin in the same
pattern as it affects all internal organs. Age-relating skin changes and its preven-
tion as a dream of human being strongly associated with loss of elasticity, epi-
How to cite this paper:
Sukoyan, G.,
Tsivtsivadze, E
., Golovach, V., Kezeli, T.
and
Demina, N. (2018) Anti-
Aging Effect
of
Cynara cardunculus
L.
var.
Cynara sc
o-
lymus
L. Extract in D-Galactose-
Induced
Skin Aging Model in Rats
.
Pharmacology &
Pharmacy
,
9
, 428-439.
https://doi.org/10.4236/pp.2018.910032
Received:
October 12, 2018
Accepted:
October 23, 2018
Published:
October 26, 2018
Copyright © 201
8 by authors and
Scientific
Research Publishing Inc.
This work is licensed under the Creative
Commons Attribution
International
License (CC BY
4.0).
http://creativecommons.org/licenses/by/4.0/
Open Access
G. Sukoyan et al.
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10.4236/pp.2018.910032 429
Pharmacology & Pharmacy
dermal atrophy, wrinkling of the skin and cause by hormonal (estrogens and
androgens) rearrangement inversed turnover collagen type I/III and collagen
degradation in dermis, dryness, worsening of the texture and appearance of skin
[1]. Skin aging is inevitably associated with a disturbance in collagen metabolism
(adult skin is composed of type I (80% - 85%) and type III collagen 10% - 15%)
due to the decreased activity of fibroblasts and their collagen synthesis, as well as
elastin [2]. Thus, the search of new medicinal technologies modulating metabol-
ic processes in the skin and improving skin resistance to oxygen demand and
oxidative stress development (the oxidative stress is one of the major mechan-
isms for skin aging and dermatological conditions [3]) and preventing the colla-
gen isoforms imbalance, is an urgent problem in dermatology. A significant
amount of evidence has been pointed to the beneficial effect of natural plants
extracts rich in phenolic and flavonoid compounds, on cellular nucleus and col-
lagen metabolism disturbances, reducing the amount of free radicals in dermal
fibroblasts [4]. Such substances are present in extracts of artichoke
Cynara sco-
lymus
L. var.
Cynara cardunculus
L.,
Asteraceae). The artichoke is emerging like
a promising medicinal plant and ancient crop in Italy, Spain, France, Egypt and
now in Georgia (Italian cultivars var.,
Cynara cardunculus
L., Grosso Romanes-
co) with favorable climatic condition. Extracts of artichoke on the tissue level
have been found to decrease the production of reactive oxygen species, the oxi-
dation of low-density lipoproteins and lipid peroxidation, protein oxidation and
increase the activity of glutathione peroxidase (for review see [5] [6] [7] [8]).
Among health-promoting properties, there were identified and characterized
antioxidant, hepatoprotective, choleretic, hypoglycemic, hypocholesterolemic,
hypotriglyceridaemic, cardiovascular, antifibrotic, anti-inflammatory and anti-
microbial, anti-dyspepsia and prebiotic effects [8]-[17]. Hepatoprotective effect
of artichoke leaf extract has a cholesterol reducing property, decrease oxidation
of low density lipoproteins throughout in part by facilitating bile production as-
sisted fat digestion together with toxin removal [18] [19]. The lipolytic action of
Cynara scolymus
L. occurs by enhancing the synthesis of the NAD-NADH2 and
NADP-NADPH2 coenzymes involved in reducer anabolic processes that are
mainly used in fatty acid synthesis in the adipose tissue and the liver. Appropri-
ate formulation of luteolin [17], sylimarin [20] and cynaropicrin [21], as com-
ponents of artichoke extracts, promising to candidate to target local and system-
ic treatments of inflammatory skin diseases [20] [22], however, little known
about direct ability of water extracts of artichoke to improvement in skin condi-
tions and skin inflammation in aging. The aim of the present study is to assess
the mechanism of therapeutic effects of lyophilized water extracts of drying
leaves of
Cynara cardunculus
L. on markers of collagen metabolism and inflam-
mation in skin in experimental animals with D-galactose induced skin aging.
2. Materials and Methods
2.1. Plant Materials and Extraction
The experiment was carried out during the 2016-17 years, in a representative
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area for
Cynara cardunculus
L. (Grosso Romanesco) var.
scolymus
L., femaly
Aesreraceae) globe artichoke cultivation in Mtskhetis region (Rosenthal, Geor-
gia, latitude 41˚56'02"N and longitude 44˚34'36"E), average minimum tempera-
ture −1˚C and maximum 35˚C. Preparation for comparison: pure Artichoke ex-
tract, 2%, ampoule 5 ml, from “Veluderm International S.L.”, Spain and 2% Ar-
tichoke extract, manufacturer “Group Companies Martinex”, Russia. The fresh
leaves of the artichoke were collected at harvest maturity from the June to the
middle of October. The leaves were separated washed, cleaned, and drying in
according with Eur Ph monograph 01/2008:1866 corrected 6.0. Extraction of
dried leaves artichoke, separation and identification of Artichoke (
Cynara car-
dunculus
L.) performed in according with Eur Ph monograph 01/2009:2389
(content of chlorogenic acid < 2.5%). Crude aqueous extracts of dried leaves
(100 g) were prepared by infusion with distilled water (plant:solvent ratio of 5:1)
at 96˚C to the homogenized leaves for 120 min, and extraction for four hours
using bi-distillated water as a solvent. Prepared extracts filtered through a metal-
lic mesh to remove any kind of solid particle, cooled at room temperature and
centrifuged at 5000 - 6000 rpm (revolutions per minute) for 15 min. The ob-
tained primary extract was filtered throughout closed sterile filtration systems
with 0.45 μ and 0.2 μ. After sterile filtration extracts concentrated by lyophiliza-
tion with a FTS Systems Lyostar II LYOACC3P1, USA lyophilizer (initial tem-
perature of −30˚C, the time of lyophilisation 24 h, additional drying at 32˚C for
6 h), previous freezing at −55˚C in a Ultra-Low-Temperature Freezer VT 407
(Vestfrost Solutions, Danish). The resulting yields were 13.7 g for dry leaf water
extracts. The toxicity of artichoke extract is very low (LD50 for mice with intra-
peritonially application is more than 2.0 g/kg body and in rats is more than 1
g/kg body weight. No microscopic lesions registries in skin of Wister rats after
application of artichoke extract twice daily, for 14 days, at concentration up to
10% and moderately irritating when applied to intact and abraded rabbit skin.
Pilot patch test on 7 volunteers (occlusive patch 48 hours) documented absence
of irritant reactions at 15 min and 24 hours after patch removal and indicated on
very good skin compatibility.
2.2. Determination of the Total Phenolics, Flavonoids and
Antioxidant Capacity (AOC)
Total phenols (TPC) was determined spectrophotometrically by measured the
absorbance at 765 ± 2 nm following the Folin-Ciocalteu reagent assay using gal-
lic acid for the preparation of calibration curve (12.5, 25, 50, 62.5, 100 and
125 mg/L). Dilutions of lyophilized water extract of artichoke leaves ranging
from 4.25 to 0.02 mg/ml were used. The total phenolic content expressed as mg
gallic acid/100 mg of extract. Total flavonoid content of was determined by the
aluminium chloride method using quercetin as a standard [23] and results were
expressed as mg of quercetin equivalents measured by absorption at 420 ± 2
nm/100 mg of extracts [24]. Total antiradical activities was determinate by eval-
uation of 1,1-diphenyl-2-picrylhydrazyl (DPPH) methods. 1 ml extract of arti-
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choke was mixed with in 4 ml of 0.004% solution of DPPH in 80% methanol (a
radical at the concentration of (working solution 24 mg/L (60.8 μmol/L) DDPH
in 80% methanol, an absorbance of 0.8 at 517 ± 2 nm) was prepared for cali-
brated curve preparation. The sample was incubated for about 60 min in dark at
22˚C and then absorption at 517 ± 2 nm was measured. AOC was calculated as
IC50 (quantity of extract neutralizing 50% of DDPH amount).
2.3. Animal Study Design
Experiments carried out in 72 female Wistar rats weighing 180 - 200 g, 170 - 180
days old. Animals received humane care in compliance with “Guide for the Care
and Use of Laboratory animals” (National Institutes of Health publication 86-23,
Revised 1996) and was performed with approval of the local Interinsitutional
(International Scientific Centre of Introduction of New Biomedical Technology,
Department of Medical Pharmacology and Pharmacotherapy, Tbilisi State Med-
ical University, Tbilisi) Animal Care and Use Committee. All animals were se-
cured under specific pathogen free conditions according to the Federation of
European Laboratory Animal Science Associations guidelines in humidity- and
temperature-controlled environment, with a daylit environment for at least 1
week before the experiments. Animals were fed commercial laboratory rat’s food
pellet and allowed drink tap water
ad libitum
before the experiments. After 7
days of adaptation, all animals randomized into two groups: control I and main.
Animals in main group after randomization received injection with D-galactose
(reducing sugar, is a naturally occurring substance in the body, 150 mg/kg/day,
i.p. [25] [26]), while in control I group received saline injection (1 mg/kg/day,
i.p.), for 8 weeks. At 21 days after injection with D-galactose the 3 cm round tat-
too area was prefabricated on each side of rats previously disinfected hip under
sterile condition and general anesthesia with pentobarbital (40 mg/kg). All ani-
mals in main group (40 animals) were secondly randomized into 4 groups in
dependence to treatment (twice in week of intradermal injection under general
anesthesia) for 5 weeks: control II group animals treated with microinjection of
saline, main I group treated with 0.12 ml of 2% lyophilized powder of Artichoke
extracts salivated in water for injection, Georgia (equivalent of average intra-
dermal dose for patients 10 mg), n = 12; main II – 0.12 ml of pure Artichoke ex-
tract, 2% (Spain), n = 12 and main III – 0.12 ml of 2% artichoke extracts (manu-
facturer “Group Companies Martinex”, Russia), n = 12. After the experiments,
all the rats were euthanized by pentobarbital (60 mg/kg intraperitonially).
2.4. Body Weight and Skin Oedema Evaluation
The mean body weight of all the animals was taken from the start of the study, 8
weeks and on the end of experiments. Appearance of skin, fur and eyes were ob-
served during these days. Skin oedema was measured as decreases in skin (1
cm2) and calculated by dividing the wet-to-dry weight ratios after tissue drying
to constant weight at 80˚C.
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2.5. Total Collagen (Hydroxyproline) and Hexosamine Contents
and Its Fraction
Immediately, skin a weighted amount of skin (3 g) was shaved, cleaned from the
underlying connective, muscular and adipose tissues was frozen in nitrogen and
minced. Lipids were removed with chloroform:methanol (2:1) mixture to deter-
mine the amount of hydroxyproline and hexasamine [27]. Total content of col-
lagen was measured assuming a hydroxyproline content of 0.91 mmol∙1−1 (mg
collagen)−1 or by multiplying the hydroxyproline content by a factor 7.46. Solu-
bility pattern of collagen was determined as described by [28].
2.6. Preparation of Nuclear Extracts and Determination of NF-kB
Skin sample frozen in nitrogen, minced in 10 volumes of solution 1, containing
10 mmol/L KCl, 10 mmol/L HEPES-buffer, 0.1 mmol/L ethylene glycol tetraa-
cetic acid (EGTA) and 0.1 mmol/L ethylenediaminetetraacetic acid (EDTA), 0.1
mmol/L phenylmethylsulfonyl fluoride (PMSF) nd 0.1 mmol/L dithiothreitol
(DTT), pH 7.8. IGEPAL (octylphenoxypolyethoxyethanol, Sigma Aldrich) was
added to a final concentration 0.7%. Skin homogenate were centrifugated twice
at 7000 g for 30 s after resuspended in 1 ml of new portion solution 1. Then pel-
let was resuspended in 0.3 ml cold solution 2 containing: 0.4 mol/L NaCl, 20
mmol/L HEPES-buffer, 1.0 mmol/L EGTA and 1.0 mmol/L EDTA, 1.0 mmol/L
PMSF and 1.0 mmol/L DTT, pH 7.8, left on ice for 20 min and the pellet of cen-
trifugated at 10,000 g for 30 min was used in the study. The activity of
NF-kB(p60) was registrated as described early [29]. The Bradford method
(Coomassie brilliant blue G-250 staining) was used to detect protein content.
2.7. Statistical Analysis
All variances in the measurement data expressed as mean ± standard deviation
of mean (SD), and statistical significance assessed using Student t-test for nor-
mally distributed variables and
p
< 0.05 is considered significant. All statistical
calculations were performed using the Statistical Sciences (SPSS, version 23.1).
3. Results
3.1. Properties of the Studying Extracts of Artichoke (Cynara
cardunculus (Grosso Romanesco), Subsp Cynara scolumus L.)
The studying water dry extracts of artichoke (
Cynara cardunculus
, cultivated in
Georgia, Mtskhetis region), 2% Pure Artichoke extract from Spain and Russia
related to water artichoke extracts with content of chloroagenic acid < 2.5%
(Assessment report on
Cynara scolymus
L., folium EMA/HMPC/150209/2009)
and about 10% of total phenolic acids. No significance difference in the contents
of total phenolic, flavonoids compounds and total antioxidant activities in the
extract made in our laboratories and VD Pure Artichoke extract, 2% (Spain)
eliminated. While values of all parameters of Artichoke extract, 2% (Russia) are
smaller (Table 1).
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Table 1. Comparison of phenolic, flavonoids content and total antioxidant activity of
different 2% extracts of artichoke.
Properties
Liophylized
extract artichoke,
Georgia
Pure Artichoke
extract, 2%
(Spain)
Artichoke extract,
2% (Russia)
Chlorogenic acid, mg/100 mg extract 1.97 ± 0.18 1.86 ± 0.14 1.25 ± 0.14**#
Total phenolic compounds, %* 9.6 ± 0.6 8.8 ± 0.8 7.7 ± 1.1*
Flavonoids, % 1.45 ± 0.09 1.51 ± 0.10 1.23 ± 0.10*#
Total antioxidant activity,
50% inhibitory of DDPH
14.3 ± 1.8 11.4 ± 1.2 8.4 ± 0.8*#
Note: values are mean of 5 samples of 5 times production of extracts and in each samples determination
performed in triplicate in case of “Lyophilized extract artichoke, Georgia” and values of 4 bath of commer-
cial Artichoke extract (Spain) and 5 bath of Artichoke extract (Russia); *- total phenols in absolute value in
“Lyophilized extract artichoke, Georgia” is 0.29 mg GAE/100 mg extract; *- compared with “Lyophilized
extract artichoke, Georgia”, # - with Artichoke extract (Spain), significance of between the extracts, signi-
ficance of difference of comparison: one symbol – p < 0.05, two – p < 0.01, absence of symbol indicated that
differences is not significance (p > 0.05).
3.2. Beneficially Therapeutic Effect of Different Artichoke
Extracts on Maintenance Level and Balanced
Soluble/Insoluble Collagens in D-Galactose-Induced
Skin Aging of Rats
Animals with D-galactose induced skin aging during 12 weeks demonstrated
evident symptoms of aging, including slow movement, a lag in response, lis-
tlessness, and withered and lackluster fur, decreased behavioral activity, some
hypodynamia, neurological impairment, established hair color changes and an
unique skin appearance, with wrinkling’s and furrows indicative of skin aging
appearing at the age (Table 1). Prior to euthanized, no morbidity/mortality and
clinically relief differences in food intake and water consumption in subgroups
of main group were not observed. The mean body weight of animals in treated
group when compare with mean body weight of animals in control III groups
showed no significant alterations in the weight gain, but was decreased in all
groups in comparison to mean body weight in first day and to control II animals
the same age as in main group (Table 2).
However, the relative weight of skin markedly decreases in D-galactose model
of aging. Water content in the skin is presumed to be a critical determinant in
cutaneous aging. Treatment with all preparation of artichoke extracts restores
the water dysbalanced in the aging skin in the same manner. Treatment with
lyophilized extract artichoke, Georgia and pure Artichoke extract, 2% (Spain)
and in less extent in case of 2% artichoke extracts manufacturers by “Group
Companies Martinex”, Russia leads to increase the content of hydroxyproline in
aging skin which compared to aging groups without treatment (Table 2). In the
samples of skin from animals of control group III occurs decreased fractions of
solubility in neutral salt and acid collagen while the collagen fraction solubility
with pepsin increased about two 2-fold. Local prolonged treatment with arti-
choke extract preparation improved solubility of skin collagen in neutral salt and
acid as compared with D-galactose treated group, meaning that the content of
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Table 2. Body weight, relative weight of skin, hydroxyproline, hexosamine and content and NF-kB activity in skin in control and
experimental animals.
Groups/Parameters
Control I,
n = 12
Control II,
n = 12
Main group, = 48
D-galactoside induced aging group
Control III
I
II
III
Body weight, g 180 days at the
beginning of the studying
186 ± 19 298 ± 23x 257 ± 18 277 ± 17 270 ± 21 259 ± 21
Relative weight, mg dry/100 mg wet weight 31.4 ± 1.4 32.1 ± 1.2 24.4 ± 1.5** 28.9 ± 1.4# 28.0 ± 1.6# 27.1 ± 1.5#
Hexosamine mg/g wet tissue 16.4 ± 1.4 18.2 ± 1.2 23.7 ± 2.3* 17.8 ± 4.1# 19.3 ± 4.3# 21.8 ± 1.2*
Hydroxyprolinemg/g wet tissue 15.0 ± 0.8 17.2 ± 0.9x 13.7 ± 0.8** 15.9 ± 0.9# 15.4 ± 1.1# 14.8 ± 1.2*
Total collagen, mg/g wet tissue 112 ±10 129 ± 10 102 ± 6* 119 ± 8# 115 ± 8# 111 ± 9
NaCl-Soluble collagen, mg/g wet tissue 7.1 ± 1.1 7.7 ± 1.0 4.4 ± 0.8** 6.5 ± 0.5## 6.35 ± 0.63## 5.97 ± 0.78#
Acid-Soluble collagen, mg/g wet tissue 0.12 ± 0.02 0.13 ± 0.03 0.07 ± 0.01** 0.10 ± 0.01*# 0.10 ± 0.01*# 0.09 ± 0.01*
Pepsin-Soluble collagen, mg/g wet tissue 0.85 ± 0.08 0.88 ± 0.08 1.8 ± 0.2*** 0.95 ± 0.10### 1.09 ± 0.09### 1.47 ± 0.15#
Insoluble collagen, mg/g wet tissue 105 ± 9 121 ± 9 97 ± 9* 113 ± 6# 109 ± 6 105 ± 7
NF-kB(p65) activity, o.u. at 450 nm 0.07 ± 0.02 0.09 ± 0.01 0.21 ± 0.03 0.10 ± 0.02 0.10 ± 0.03 0.14 ± 0.02
Note: control II practically healthy animals of control I group (170 - 180 days old) at 230 - 240 days old as in main groups. *- compared with control II, # -
with control III, §- main I group, ∆ - main II group, x – between control I and control II; significance of between the extracts, significance of difference of
comparison: one symbol – p < 0.05, two – p < 0.01, absence of symbol indicated that differences is not significance (p > 0.05).
newly synthesized collagen had been increased and its level is known to decrease
with age in skin [30]
.
Early it was shown that soluble collagen extractable by 0.5
M NaCl showed rise after birth, reach maximum at the age of 8 weeks and then
decline with animal aged. While level of insoluble collagen in skin rose conti-
nuously over the 12 month and revealed a strong correlation with tensile
strength. Alternatively, the increase of hexosamines up to 23.7 ± 2.3 (vs. 18.2 ±
1.2 mg/g wet tissue in the control) in D-galactose induced aging skin indicated
of presence of some signs of inflammation. Decrease the absolute levels of hex-
asamines and ratio between collagen (hydroxyprolyne): hexosamine under
treatment with artichokes extracts could indicate to elimination of on acute fi-
brosis processes.
3.3. Changes of Inflammatory Markers in D-Galactose-Induced
Skin Aging and Efficacy of Artichokes Extracts Action
Several studies point to a role for NF-κB in modulating epidermal thickness and
apoptotic susceptibility of fibroblast [31] and keratinocytes [32] [33]. In
D-galactose subacute model, skin aging activity of NF-kB (p65) increases by
133% (Table 2). Benefit for positive action of artichokes extracts inflammation
confirmed by decreasing the activity of NF-kB (p65) by 50% up to level charac-
terized to skin rats at the same age.
4. Discussion
The present study designed to investigate the potential protective effect of
low-dose Georgian lyophilized extracts of artichoke against the skin markers of
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elasticity caused by inversed turnover of collagen type ratio I/III dysbalance in
D-galactose induced skin aging model in rats. Skin aging is a complicated multi-
targets dysbalancing progression in the epidermal and dermal layers resulting
from many intimal intrinsic and extrinsic factors. The major topic of research of
pathogenesis and molecular mechanism of skin aging focused on the main der-
mal constituent collagen (about 60% - 80% of skin-mass [33]) and ability to
maintenance collagen regulatory system, to prevent a loss of the skin’s ability to
resist stretching and degradation in aging. Artichoke extracts, a preparation
which represents a natural source of phenolic compounds with dicaffeoylquinic
acid along with their precursor chlorogenic acid (5-caffeoylquinic acid) as the
predominant molecules has been traditionally used as a hepatoprotective, di-
uretic, detoxifying, lipid lowering agents with antioxidant effects and ability
to reduce postprandial blood glucose (for review see [5] [6] [7] [8]). Pre-
treatment of
Cynara scolymus
extract in N-nitrodiethylamine and carbon te-
trachloride-induced hepatocarcinogenesis in rats limited not only oxidative
stress production, but as much important, decrease level of proinflammatory
cytokines TNF-alpha and nuclear transcription factor-kappa B (NF-kB) [20]
[22]. Canonical NF-κB pathways signaling has a crucial role in the maintenance
of skin homeostasis [34]. Chronic, low-grade inflammation is also recognized as
a major characteristic of the aging process [35]. This phenomenon is called “in-
flammaging”. Given that the mechanism of skin “inflammaging” is far from be-
ing thoroughly understood, little progress is made to develop targeted treat-
ments. Suppression activity of NF-kB included in the intrinsic mechanism of in-
hibition of tumor cell proliferation and apoptosis in N-Nitrosodiethyleamine in-
duced hepatic injury [20] and blocking NF-kB activity could reverse effects of
advanced glycation end-products on cell proliferation and migration, but not
adhesion [36]. Previous studies also suggest that the NF-kB pathway plays an es-
sential role in psoriasis progression [37] [38], where there is marked elevation of
active phosphorylated NF-kB p65 in psoriases, as an inflammatory skin disease
in which activated immune cells and the pro-inflammatory cytokine TNF are
well-known mediators of pathogenesis [17]. Luteolin could be inhibiting in-
flammatory mediator production via blockade of NF-kB activation. However, it
is also possible that luteolin acts on signaling molecules upstream of NF-kB ac-
tivation, such as the mammalian target of rapamycin (mTOR) that governs cell
size, growth, and metabolism [39]. Which of following or other molecular me-
chanism is responsible for undergoes collagen metabolism changes under
D-galactose induced subacute skin aging and is it the same with normal skin ag-
ing processes is subject to a further research.
5. Conclusion
In conclusion, the results of the current study demonstrated the anti-aging effect
of black seed oil in a mouse model of aging induced by D-galactose. Obtained
data indicate that the concomitant use of 2% artichoke extract could be the only
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way to exert anti-aging action in this model of skin aging in experimental ani-
mals, way to prevent imbalance in collagen metabolism and de-remodeling
function of aging skin. Influence of artichoke extracts restored skin relative
weight and led to an increase of solubility in neutral salt and acid and decreased
pepsin solubility collagen fraction, restored the hexosamine/collagen (hydroxy-
proline) ratio and decreased the activity of nuclear transcription factor (NF-kB).
Local prolonged treatment with artichoke extracts improved collagen metabol-
ism and attenuated the progression of inflammation in D-galactose induced skin
aging model.
Acknowledgements
We thank the Chief Kamkamudze R. and his staff for animal housing.
Funding
The authors received no specific funding for this work, article written indepen-
dently; no company or institution supported it financially.
Conflicts of Interest
The authors declare that they have no conflict of interests regarding the publica-
tion of this paper. The authors alone are responsible for the content and writing
of this article. No competing financial interest or financial conflict with the sub-
ject matter or materials discussed in the manuscript exists.
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