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1Departemen nt of Medicine Faculty of Medicine University of Prima Indonesia Meda
4Departement of Biomedical Science Faculty of Medicine University of Prima Indonesia Medan
Antioxidant and Anti-Hyaluronidase Activities of Dragon Fruit Peel Extract and Kaempferol-3-O-
Rutinoside
Aktivitas Antioksidan dan Anti-Hialuronidase Ekstrak Kulit Buah Naga dan Kaempferol-3-O-
Rutinoside
Liana Liana , Rizal Rizal , Wahyu Widowati , Fioni Khainir Akbar , Edy Fachrial , I Nyoman Ehrich L
1 2 3 1 1 1 4
,
2Biomolecular and Biomedical Research Center Aretha Medika Utama Bandung
3Faculty of Medicine Maranatha Christian University Bandung
ABSTRACT
Keywords: ging anti-hyaluronidase, ragon fruit peel extract, kaempferol,H O scavengingA , d 2 2
Kata Kunci: Antihialuronidase,ekstrak kulit buah naga, pemerangkapan H Okaempferol-3-O-rutinoside, 2 2
Aging is a natural process in human life and is triggered by the presence of free radicals (ROS). The use of antioxidants from
natural ingredients is one of the breakthroughs to overcome aging and counteract the harmful effects caused by the free
radicals. This study aimed to determine and compare the antioxidant activity of H O scavenging and hyaluronidase
inhibition of red dragon fruit peel extract (DFPE) and kaempferol-3-o-rutinoside (KOR) compounds. ragon fruit peel
extract DFPE is obtained through extraction by maceration method using 70% ethanol solvent. The design of this study
included antioxidant and anti-aging activity assay of EKBN and KOR at the series concentration of 15.63; 31.25; 62.50; 125;
250; 500 µg/mL through H O scavenging, as well as the DFPE and KOR hyaluronidase inhibition assay at the series
concentration of 5.21; 10.42; 20.83; 41.7; 83.33; 166.67 µg/mL. EKBN shows that the average activity of H O scavenging is
lower than KOR. In addition, the IC values of KOR for H O scavenging is lower (351.46±2.30ug/mL) than DFPE
(409.64±23.17ug/mL). While, KOR also has higher values of inhibitory activity than of the DFPE. However, the IC value of
KOR for hyaluronidaseinhibition activity was 84.07±10.46µg/mL, equivalent to the IC value of DFPE (85.32±10.24µg/mL).
The presence of antioxidant and anti-aging activity in the EKBN is probably caused bybetalain and the KOR compound itself
contained in red dragon fruit. The results of the paired-samples T-test on antioxidant activity and anti-aging of DFPE and
KOR showed non-significant difference. Thus, DFPE has an equivalent antioxidant and anti-aging through H O scavenging
and hyaluronidase activity as possessed by the KORcompound.
2 2
D
( )
2 2
2 2
50 2 2
50
50
2 2
ABSTRAK
Penuaan merupakan proses alamiah dalam kehidupan manusia yang salah satunya dipicu oleh keberadaan radikal bebas
(ROS). Penggunaan antioksidan dari bahan alami merupakan salah satu upaya untuk mengatasi penuaan dan menangkal
efek bahaya yang ditimbulkan oleh radikal bebas. Penelitian ini bertujuan untuk menentukan
aktivitas antioksidan pemerangkapan H O dan penghambatan hialuronidase ekstrak kulit buah naga
(EKBN) dan senyawa (KOR). Ekstrak kulit buah naga merah (EKBN) kstraksi
dan membandingkan
2 2 anti-aging merah
kaempferol-3-o-rutinoside didapatkan melalui e
yang menggunakan . Desain penelitian ini meliputi aktivitasdilakukan metode maserasi dengan pelarut etanol 70% uji
antioksidan , , , pemerangkapan H O
dan penghambatan , , , , ,
EKBN dan KOR pada seri konsentrasi 15 63; 31 25; 62 50; 125; 250; 500µg/mL melalui 2 2
serta uji EKBN dan KOR pada seri konsentrasi 5 21; 10 42; 20 83; 41 7; 83 33;
166 67µg/mL EKBN menunjukkan rata-rata aktivitas pemerangkapan H O yang lebih rendah dibandingkan KOR. Selain
itu, perhitungan n menunjukkan
hyaluronidase
, . 2 2
ilai IC pemerangkapan H O KOR lebih rendah (351,46±2,30ug/mL) dibandingkan EKBN
(409,64±23,17ug/mL).
50 2 2
Sementara KOR juga mempunyai rata-rata aktivitas penghambatan yang lebih besar dibandingkan
EKBN. Namun, n penghambatan ,ilai IC KOR pada aktivitas hialuronidase adalah 84,07±10,46µg/mL setara dengan nilai
IC EKBN (85,32±10,24µg/mL).
50
50 Adanya aktivitas antioksidan dan pada EKBN diduga disebabkan oleh betalain
dan senyawa KOR itu sendiri yang terkandung pada buah naga merah. Hasil uji terhadap aktivitas
antioksidan dan EKBN dan KOR menunjukkan perbedaan aktivitas yang tidak signifikan. Dengan demikian,
anti-aging
paired-samples T-test
anti-aging
EKBN mempunyai kapasitas antioksidan pemerangkapan H O dan penghambatan aktivitas hialuronidase yang
setara dengan senyawa KOR.
2 2 anti-aging
Korespondensi: Liana . Departemen Faculty of Medicine University of Prima Indonesia Medan, Jl. Belanga No. 1, Sei
Putih Tim. I, Medan Petisah, Medan, Sumatera Utara 20111 Indonesia Tel. (061) 4578870/08126210328 Email: lianacth@gmail.com
Liana t of Medicine
Jurnal Kedokteran Brawijaya Vol. 30, No. 4, Agustus 2019, pp. 247-252
Artikel Penelitian
Article History: Received 11 Maret 2019, Accepted 26 Mei 2019
Dapat diakses pada: http://jkb.ub.ac.id/index.php/jkb/article/view/2546
DOI: http://dx.doi.org/10.21776/ub.jkb.2019.03 .0 .0 4 3
247
248
INTRODUCTION
Biodiversity of Indonesia provides variety of products that
contain antioxidant activities, reddragon fruit (Hylocereus
polyrhizus) is one of them. Besides utilizing the fruit, it
turns out that its peel can also be processed into basic
ingredients for cosmetics making and natural food
coloring. This is because the dragon fruit peel contains
many compounds that can be used as antioxidants
including betalain and anthocyanin. In addition, dragon
fruit peel also contains vitamin C, Vitamin E, vitamin A,
alkaloids, terpenoids, flavonoids, thiamine, niacin,
pyridoxine, cobalamin, phenolic, carotene, and phyto-
albumin . (KOR) is one of the
flavonoids found in dragon fruit . Many studies have
been conducted to determine the benefits of KOR such as
ant i oxidant s , a nti-inf l a m m atory, ant i m i c robia l ,
anticancer, heart disease prevention, neurological
diseases, antidiabetic, antiosteoporosis, antiestrogenic,
analgesic, and hypo-allergenic. In addition to antioxidants
compounds, red dragon fruit also found to have anti-aging
properties. KOR is also a strong compound that can inhibit
hyaluronidase enzymes .
(9) Kaempferol-3-o-rutinoside
(10)
(11)
H O Scavenging Activity Assay
2 2
Preparation of Dragon Fruit Peel Extract (DFPE)
Hyaluronidase Inhibitory Activity Assay
METHOD
Radical scavenging activity of H O was measured based on
a method modified by Mukhopadhyay .
Previously Dragon Fruit Peel Extract (DFPE) was obtained
from extraction and (KOR) was
purchased from Chengdu Biopurify Phytochemicals Ltd.
(Chengdu, BP0823) The mixture of the solution was made
by adding 60 L of various concentrations of extract sample
and compounds (15.63 500µg/mL) into the sample and
blank well. As much as 12 L of Ferrous Ammonium Sulfate
(1 mM, Sigma 7783859) was added into the control and
sample, H O solution (5mM, Merck 1.08597.1000) was
added to the sample well only, 63 L 10% DMSO was added
into control well, and 90 L 10% DMSO into blank well. The
mixture was incubated in a dark room at room
temperature for 5 minutes. Each sample and control well
was added with 75 L 1,10-phenanthroline (1mM, Sigma
131377). The plate was then incubated for 10 minutes in a
dark room and at room temperature. The absorbances
were measured using spectrophotometry at 510 nm
wavelength The percentage of scavenging
activities was calculated using this following formula:
2 2
et al.(15)
,
Kaempferol-3-o-rutinoside
.
μ
-
μ
2 2
μ
μ
μ
(12-14).
Aging is a natural process in human life and closely related
to various regenerative processes Intrinsic and
extrinsic factors influence the aging process in skin tissue
Intrinsic aging is an aging process that cannot be
prevented and occurs over time at cellular level. Intrinsic
aging is affected by changes in sex hormone production
associated with increased age. The biological process
plays a role in determining the amount of multiplication in
each cell until the cell stops dividing and then dies.
Extrinsic aging is caused by an exposure to external
factors, especially UV light. Thus, it is often referred to as
photoaging. The other factors of extrinsic aging include
pollution, smoking habits, and unbalanced nutrition
(1).
.
(2,3).
Hyaluronidase inhibitory activity was measured by a
modified method from Sigma Aldrich and Tu and Tawata
(12-14,16). A mix of 25 L of various concentrations of
sample and compound extract (5.21-166.67 g/mL), 3 L
hyaluronidase from bovine testes type I-S (Sigma H3506,
USA), 12 L phosphate buffer (300 mM, pH 5.35) was
incubated for 10 minutes at 37°C. Then, 10 L hyaluronic
acid substrate (Sigma H5542, USA) was added and
incubated for 45 minutes at 37°C. The reaction was
stopped by adding 100 L acidic albumin acid. The mixed
solution was then incubated at room temperature for 10
minutes. The absorbance was measured at 600 nm
wavelengths . The inhibition activity was calculated
using the following formula:
μ
μ μ
μ
μ
μ
(12-14)
This study aimed to determine the antioxidant activity and
anti-aging of dragon fruit peel extract and make a
comparison with KOR compound through the analysis of
H O scavenging and hyaluronidase inhibition assay. Thus,
DFPE which has been a waste can be utilized as an
antioxidant and anti-aging agent derived from natural
products.
2 2
The production of ethanol extract was carried out by the
maceration method using 70% ethanol solvent. An amount
of 500g of dried red dragon fruit peel that had been sliced
and placed into a glass container was added with 3.75L
ethanol 70% and then covered using the lid. The sample
was left for five days and protected from light. It was then
wrapped using flannel cloth and stirred until a liquid
extract solution was obtained. The filtered pulp was
macerated again with 1.25L ethanol solution. Then, the
second step of filtration was conducted, and the filter
result was combined with the first result. The result
obtained was concentrated with a rotary evaporator until
all the solvents evaporated and followed by the
evaporation process above water bath until obtaining a
paste-form extractof dragon fruit peel (12-14).
Currently, many research are conducted to prevent skin
aging. The use of antioxidants is one of the efforts to
overcome skin aging besides the use of anti-aging
cosmetics because antioxidants can counteract the
harmful effects caused by free radicals. In addition to
antioxidants, some inhibitor compounds for extracellular
matri x enzy mes d egradat ion s uch as e lastase ,
hyaluronidase, and collagenase have been known to play
an important role in preventing skin aging .
Hyaluronidase in h i b i t o r s w o r k b y p r e v e n t i n g
hyaluronidase activity which degrades hyaluronic acid.
Hyaluronic acid is an important component of Extra
Cellular Matrix (ECM) that plays a role in maintaining skin
moisture and its elasticity. There are many cosmetics
containing synthetic hyaluronic acid to prevent skin aging
available world-wide. However, people or scientist tend to
develop natural cosmetics due to their minimum safety
risks and side effects comparedto the synthetic one .
(7)
(8)
The existence of free radicals due to oxidative processes is
a mechanism mostly used to explain the occurrence of the
aging process. Free radicals are molecules that have one
or more unpaired electrons in their outer orbit, so they are
relatively unstable or often referred to as Reactive Oxygen
Species (ROS) . These molecules have an action
mechanism to take electrons from various vital
components of cells, such as DNA, cytoskeleton, protein,
and cell membranes, and cause damage at the cellular
level and accelerate the aging process
(4,5)
(6).
Jurnal Kedokteran Brawijaya, Vol. 0, No. 4, Agustus 2013 9
Antioxidant and Anti-Hyaluronidase Activities of Dragon....
RESULTS
Table 1. Scavenging activity of H O by DFPE and KOR
2 2
Statistical Analysis
Hydrogen peroxide (H O ) is a free radical that is found in
nature. The scavenging activity can be measured by the
re a c ti on o f f e rr o us a mm on iu m su lf a t e an d
phenanthroline. The complex reaction will produce an
orange complex of Fe -tri-phenanthroline. The presence
of H O inhibits the formation of complex, but in the
existence of antioxidants that scavenged H O , the complex
will be formed again. Thus, the orange color formation
indicates the occurrence of H O scavenging activity .
2 2
2+
2 2
2 2
2 2 (15)
Data from the results of H O scavenging and anti-
hyaluronidase activities were analyzed using One-Way
ANOVA and followed by the post hoc test Tukey HSD test
(IBM SPSS 22) to determine the significance between each
concentration of the DFPE and KOR. Paired Samples T-test
(IBM SPSS 22) also was performed to analyze the
significance difference betweentwo different samples.
2 2
H O Scavenging Activity
2 2
Note: Data were presented as mean ± standard deviation. Different
lowercase letters in the same column are significant at <0.05 (Tukey
HSD post hoc test).
p
The average percentage of H O scavenging activity of KOR
shown in Table 1 was higher compared to the scavenging
activity of DFPE. There is a significant difference (p<0.05)
between concentrations of both treatments, DFPE and
KOR. It implied H O scavenging activity of DFPE and KOR
compounds among concentrations of 500-15.63µg/mL
were the modes that depend on concentration (Figure 1
and Table 1).
2 2
2 2
Figure 1. Effect of various concentrations of DFPE and KOR
toward H O scavenging activity. DFPE and KOR were diluted
in DMSO 10% to reach the final concentration of 500; 250;
125; 62.50; 31,250; 15.63 (µg/mL)
2 2
Table 2. The IC value of H O scavenging by DFPE and KOR
50 2 2
Note: *Linear equations, the coefficient of regression (R2) and IC50 of
each sample were calculated.
The IC value of H O scavenging of DFPE and KOR were
shown in Table 2. The results of IC values calculation
based on the linear regression equation obtained showed
that the IC value of KOR compound was lower
(351.46±2.30µg/mL) than the IC value produced by
dragon fruit peel extract (409.64±23.17µg/mL). The
smaller the IC value, the better the ability of a compound
to scavenge the free radicals . However, based on
paired samples T-test (p<0.05) (Table 3), the whole H O
scavenging activities of DFPE and KOR were not
significantly different. It suggested, the DFPE had the same
H O scavenging activity as it was revealed by the KOR.
50 2 2
50
50
50
50
(17)
2 2
2 2
Table 3. The IC value of H O scavenging by DFPE and KOR
50 2 2
Table 4. Hyaluronidase inhibition activity of H O DFPE and
KOR
2 2
Hyaluronidase Inhibitory Activity
Hyaluronidase is one of the metalloproteinases matrixes
that function in degrading hyaluronic acid (HA), a
component in the extracellular matrix (ECM). The
mechanism of HA degradation by hyaluronidase is carried
out through the catalysis of hyaluronic hydrolysis
reactions. The hyaluronidase enzymes activity can be
detected by measuring the HA levels. HA can be measured
because of its ability to form turbidity when reacting with
albumin acid solution. The turbidity that can be captured
by a wavelength of 600nm is proportional to the HA
concentration. The inhibition activity of the hyaluronidase
enzyme is characterized by a higher HA concentration in
the remaining after the reaction is stopped. The
mechanism of hyaluronidase inhibition is a vital principle
to postpone the aging process caused by the degradation
of extracellular matrix components(12).
Assay Sample Sig. (2-
tailed)
Conclusion
H2O2Scavenging KOR –DFPE .257
Sig. (2-tailed) <0.05, H2O2
Scavenging activity between
KOR and DFPE are not
significantly different
Hyaluronidase
Inhibitory Activity KOR –DFPE .344
Sig. (2-tailed) <0.05,
Hyaluronidase Inhibitory
activity between KOR and DFPE
are not significantly different
Sample Equation R2IC50 (µg/mL)
Dragon fruit
peel extract
Y = 0.0.090x+9.700 0.960 409.64±23.17
Kaempferol-3-O-
rutinoside Y =0.122x+7.123 0.970 351.46±2.30
249
Antioxidant and Anti-Hyaluronidase Activities of Dragon....
Final Concentrations
(µg/mL)
The average of H2O2scavenging activity (%)
Dragon fruit
peel extract
Kaempferol-3-o-
rutinoside
500.00 57.05±3.39 f65.71±0.17f
250.00 36.06±0.48 e40.23±0.43e
125.00 27.98±0.49 d28.17±0.57d
62.50 18.65±0.31 c15.45±0.67c
31.25 11.14±0.54 b9.95±0.65b
15.63 5.04±0.44 a3.61±0.14a
Concentrations
(ug/mL)
The average of hyaluronidase inhibition activity (%)
DFPE Kaempferol-3O-
Rutinoside
166.67 69.75±1.52d78.06±7.32e
83.33 51.90±3.46c49.71±1.82d
41.67 42.41 ±8.01b,c 38.10±4.37c
20.83 34.41 ±4.93a,b 31.24±1.12b,c
Jurnal Kedokteran Brawijaya, Vol. 0, No. 4, Agustus 2013 9
Table 5. The IC value of hyaluronidase inhibition activity of
DFPE and KOR
50
Note: Data were presented as mean ± standard deviation. Different
lowercase letters in the same column are significant at <0.05 (Tukey
HSD post hoc test).
p
Table 4. Hyaluronidase inhibition activity of H O DFPE and
KOR
2 2
Based on the results that is shown in Table 3, the
hyaluronidase inhibition activity of KOR compound only
higher at the highest concentration compared to that of
dragon fruit peel extract. Also, there was a decrease in
hyaluronidase inhibition activity from high concentrations
to low concentrations (Figure 2). Only concentrations
between 166.67µg/mL and 83.33µg/mL of both DFPE and
KOR compounds had significantdifferences (p<0.05).
Figure 2. Effects of various concentrations of DFPE and KOR
on hyaluronidase inhibition activity. DFPE and KOR were
diluted in DMSO 10% to reach the final concentration of
166.67; 83.33; 41.67; 20.83; 10.42; 5.21 (µg/mL)
The results of calculations using the linear regression
equation is shown in Table 4. It Showed the IC of
hya l u ro ni da se i nh ib i ti on b y KO R c o mp ou nd
(84.07±10.46µg/mL) was lower than the IC value that is
produced by DFPE (85.32±10.24µg/mL). However, the
difference in IC values was relatively small It also
supported by the analysis of paired samples T-test
(p<0.05) (Table 3) that the whole hyaluronid ase inhibition
activities of DFPE and KOR were not significantly
different. Therefore, it can be assumed that DFPE had an
equal inhibitory activity to its comparative compound,
KOR.
50
50
50 .
DISCUSSION
Phenol is one of the compounds highly contained in plants
and has been widely studied due to its biological activities
such as anti-mutagenic, anticancer, anti-aging, and
antioxidant. The DFPE from is known to
contain more phenolic compounds than the other types of
dragon fruit plants. The phenolic compounds contained
and characterized in DFPE are chlorogenic acid, gallic acid,
and quercetin. These three phenolic compounds are
widely known as powerful antioxidant agents . Its
reddish color was one indication of a high content of
phenolic compounds and betalain components. Betalain is
a pigment that can dissolve in water and consists of
betacyanin (violet) and betaxanthin (yellow). Higher
antioxidant capacity was found in red dragon fruit plants,
presumably because of the high betalain content
H. polyrhizus
(18)
(19).
There are two types of antioxidants that are categorized
based on its mechanism of action, namely primary and
secondary antioxidants. The type of antioxidant capacity
examined in this study can be classified into the primary
antioxidant group. Primary antioxidants work through free
radicals scavenging mechanism and turn them into more
stable compounds or products by inhibiting initiation and
damaging the propagation chain by transferring hydrogen
atoms or electron. Whereas, the mechanism of secondary
antioxidant action is through inhibition of free radical
formation and protection against oxidation damage, which
was not examined in this study because it involves an assay
model .(21)
Kaempferol-3-o-rutinoside (KOR) is a natural flavonol
compound which is a derivative of the flavonoid group and
commonly found in fruits, vegetables, and other herbs
such as grapes, tomatoes, broccoli, and gingko biloba
leaves. In addition, KOR is one of the flavonoids found in
dragon fruit; therefore KOR was used as a comparative
compound in this study. KOR compound has high biological
activities such as antioxidants, anti-inflammatory,
antimicrobial, antidiabetic, and anticancer activities. The
flavonoid compound characteristics owned by KOR result
in high antioxidant activity .(20)
Based on the result of H O free radical scavenging activity
of DFPE and KOR compounds (Table 4.3), both showed
decent primary antioxidant activity. However, based on IC
values, the antioxidant capacities owned by DFPE (IC
409.64±23.17µg/mL) and the comparative KOR compound
(IC 351.46±2.30µg/mL) were considered as weak. In
accordance with the standards that classified antioxidant
capacity based on IC values are very strong (IC value less
than 50ppm), strong (IC value between 50-100ppm),
moderate (IC between 100-150ppm), and weak (IC
value between 151-200 ppm or more) . According to a
study conducted by Lourith & Kanlayavattanakul, IC
values were also influenced by solvents used in the
extraction process. The study compared several solvents
used in the extraction method. The results indicated that
IC value of DFPE in the ABTS and DDPH assay resulted in a
lower value, indicating a strong antioxidant in the sample
dissolved in water compared to the sample dissolved in a
polar ethanol solvent and non-polar n-hexane solvent .
2 2
50
50
50
50 50
50
50 50
(13)
50
50
(18)
Based on the DPPH assay, a study conducted by Tatsimo et
al.,showed that kaempferol was considered to have a
strong antioxidant capacity with an IC value of
52.48µg/mL. This result suggested that the antioxidant
capacity of a certain compound was proportional to IC
50
50
Note: *Linear equations, the coefficient of regression (R2) and IC50 of
each sample were calculated.
250
Antioxidant and Anti-Hyaluronidase Activities of Dragon....
Concentrations
(ug/mL)
The average of hyaluronidase inhibition activity (%)
DFPE Kaempferol-3O-
Rutinoside
20.83 34.41 ±4.93a,b 31.24±1.12b,c
10.42 26.48 ±1.62a25.05±4.56a,b
5.21 24.32 ±4.59a18.25±0.91a
Sample Equation R2IC50
(µg/mL)
Dragon fruit peel extract Y = 0.272x + 26.63 0.960 85.32 ± 10.24
Kaempferol-3-Rutinoside Y =0.345x + 21.15 0.983 84.07 ± 10.46
Jurnal Kedokteran Brawijaya, Vol. 0, No. 4, Agustus 2013 9
value and its properties and vary depending on the free
radicals that were scavenged . This allegation was also
supported by findings previously obtained by Vellosa et
al. that hydrogen peroxide (H O ) was a lesive free radical
which can cause severe lesion conditions; therefore
specific agents are needed to fight against H O . Hence,
among the 3 compounds that have been examined in the
study (quercitrin, iso-quercitrin, and kaempferol), no one
could react with H O . Another study also revealed
that anthocyanin and betalain content in DFPE are the
best for antioxidant effect. These pigments can reduce
free radicals by binding metal ions and formed a more
stable pigment-metal ion complex
(22)
,2 2
2 2
2 2 (23)
(24).
Extrinsic aging caused mainly by UV radiation can increase
the number of free radicals or ROS in the skin which can
cause inflammation and oxidative stress. This triggers the
skin aging process that is closely associated with the
overexpression of matrix metalloproteinase. The
decrease of epidermal hyaluronic acid was one of the
major changes that occur in skin aging and is found in
some skin aging cases. In addition, a decrease in skin
elasticity and premature skin aging was significantly
correlated with an increase in elastase and hyaluronidase.
Therefore, one mechanism to inhibit aging was to reduce
various enzyme activities that support matrix degradation
from the abundant connective tissues in the skin (25,26).
The antioxidant capacity of a certain compound was
closely related to its anti-aging potential. This is due to the
ROS mechanism which is one of the main causes of skin
aging. Thus, compounds having high antioxidant capacity
also have strong anti-aging activity . This was in line
with research conducted by Tu and Tawata that certain
extracts with high DPPH scavenging activity and low IC
values also showed high hyaluronidase inhibitory activity
(27)
50
(16).
Based on the results obtained in this study, the
hyaluronidase inhibitory activity by both dragon fruit peel
extract and its comparative compound were considered
as strong enzyme inhibitors with IC values of
85.32±10.24µg/mL and 84.07±10.46µg/mL respectively.
The results obtained by Tu & Tawata also showed that IC
value of an extract for anti-hyaluronidase ranging from 50
100ppm was considered as a strong inhibitor .
Comparative compound kaempferol had generally smaller
hyaluronidase inhibitory activity and IC value. Thus its
capacity as anti-hyaluronidase was higher than that of
dragon fruit peel extract. However, the difference of IC
value of hyaluronidase inhibition owned by DFPE was
relatively small. It also supported by the statistical analysis
that the whole hyaluronidase inhibition activities of DFPE
and KOR were not significantly different. Hence, it had
equal anti-aging potential to KOR, especially for
hyaluronidase inhibitions.
50
50
-
(16)
50
50
The hyaluronidase inhibition capacity was caused by
phenol and flavonoid contents found in both DFPE and
KOR. Free radicals trigger the activation of various
degrading enzymes; one of them is hyaluronidase that is
known as matrix metalloproteinase (MMP) through the
inflammatory process. The inflammation mechanism
begins with the induction of various growth factors and
proinflammatory cytokines (TNF-A, EGF, IL-1) by free
radicals. Furthermore, these cytokines carried signal
transduction and activated AP-1 protein which was a
transcription factor to increase MMP expression and
inhibited the expression of collagen. MMP worked through
the degradation of various proteins and components
found in ECM and caused the skin to lose its elasticity,
wrinkles, hyper pigmentation, and tr iggers ot her
inflammatory responses. The presence of antioxidant
compounds could prevent the ECM destruction process
through the prevention mechanism of free radical
stabilization by donating hydrogen atom or electron that
causes inhibition of MMP production. In addition,
antioxidant compounds were found to inhibit MMP work.
Thus , they were con sidered to have a nt i-agi ng
characteristics (1,5).
This study concluded that DFPE has an equivalent
antioxidant and anti-aging through H O scavenging and
hyaluronidase activity as possessed by the KOR compound.
It supported by the statistical analysis that the whole H O
scavenging and hyaluronidase inhibition activities of DFPE
and KORwere not significantly different, although generally
KORhad higher activities and lower IC in both assays.
2 2
2 2
50
ACKNOWLEDGEMENT
This research was supported by University of Prima
Indonesia, Medan, and Biomolecular & Biomedical
Research Center, Aretha Medika Utama, Bandung,
Indonesia. We thank Ni Luh Wisma, Annisa Amalia, and
Uba dillah from Biomolecular and Biomedical Research
Center, Aretha Medika Utamafor their valuable assistance.
y
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