See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/323259391

Mangosteen peel extract exhibits cellular antioxidant activity by induction of

catalase and heme oxygenase‐1 mRNA expression

Article  in  Journal of Food Biochemistry · February 2018

DOI: 10.1111/jfbc.12511

CITATIONS READS

0 266

7 authors, including:

Pattamapan Lomarat Supachoke Mangmool

Mahidol University Mahidol University
10 PUBLICATIONS   43 CITATIONS    40 PUBLICATIONS   761 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Phosphoinositide 3 Kinase View project

All content following this page was uploaded by Supachoke Mangmool on 08 August 2018.

The user has requested enhancement of the downloaded file.

Received: 8 August 2017 | Revised: 19 December 2017 | Accepted: 2 January 2018

DOI: 10.1111/jfbc.12511

FULL ARTICLE

Mangosteen peel extract exhibits cellular antioxidant activity by

induction of catalase and heme oxygenase-1 mRNA expression

Nattapon Jaisupa1,2 | Primchanien Moongkarndi3 | Pattamapan Lomarat4 |

Jutima Samer5 | Vatchara Tunrungtavee3 | Weerasak Muangpaisan6 |
Supachoke Mangmool2

1
Department of Pharmacology,
Phramongkutklao College of Medicine,
Abstract
Bangkok, Thailand Mangosteen (Garcinia mangostana L.) is one of the most popular fruits in tropical regions. Mangos-
2
Department of Pharmacology, Faculty of teen peel contains several phytochemicals that exhibits antioxidant properties. However, the
Pharmacy, Mahidol University, Bangkok, mechanisms underlying the capacity for mangosteen peel extract (ME) to inhibit cellular oxidative
Thailand
stress have not been fully defined. In this study, we found that ME significantly inhibited hydrogen
3
Department of Microbiology, Faculty of
peroxide (H2O2)-induced intracellular reactive oxygen species (ROS) production in human neuro-
Pharmacy, Mahidol University, Bangkok,
Thailand blastoma cells (SK-N-SH). Treatment with ME significantly increased the mRNA levels of catalase
4
Department of Food Chemistry, Faculty of (CAT) and heme oxygenase-1 in both SK-N-SH and human embryonic kidney (HEK-293) cells. In
Pharmacy, Mahidol University, Bangkok, addition, elevation of CAT enzyme activity was observed in ME-treated SK-N-SH cells. ME pos-
Thailand sessed significant antioxidant capacities as determined by anti-lipid peroxidation, H2O2 scavenging
5
Department of Physiology, Faculty of activity, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity assays. We found that
Pharmacy, Mahidol University, Bangkok,
(2)-epicatechin and the proanthocyanidin dimer are the active compounds in ME that elicit its anti-
Thailand
6 oxidant activities.
Department of Preventive and Social
Medicine, Faculty of Medicine Siriraj Practical applications
Hospital, Mahidol University, Bangkok,
Mangosteen peel has been traditionally used to treat some illnesses. It contains several phyto-
Thailand
chemicals, including phenolic compounds and flavonoids which elicit a variety of benefits, including
Correspondence antioxidant and cytoprotective effects. The present study demonstrated that mangosteen peel
Supachoke Mangmool, Department of
extract (ME) is a rich source of antioxidants that may have potential uses in nutraceuticals for pre-
Pharmacology, Faculty of Pharmacy,
Mahidol University, 447 Sri-Ayuthaya Road, venting diseases associated with oxidative stress.
Rajathevi, Bangkok 10400, Thailand.
Email: supachoke.man@mahidol.ac.th KEYWORDS
antioxidant activity, catalase, epicatechin, heme oxygenase-1, mangosteen peel extract,
Funding information
Ministry of Public Health proanthocyanidin

1 | INTRODUCTION demonstrated that vegetables and fruits, including mangosteen, con-

Oxidative stress is a condition of imbalance between reactive oxy-
gen species (ROS) production and cellular antioxidant defense mech-
anisms, which can cause cell injury and damage. Increased ROS
levels are associated with pathogenesis of many diseases. Protection
against oxidative stress and the breakdown products of oxidized
proteins and lipids is provided by several antioxidant enzymes such
as catalase (CAT), glutathione peroxidase (GPx), and superoxide dis-
mutase (SOD) (Apel & Hirt, 2004). Several studies have been
tains several antioxidant compounds such as phenolics, flavonoids,
quinones, and vitamins, which can reduce the oxidative stress
(Anantachoke, Lomarat, Praserttirachai, Khammanit, & Mangmool,
2016; Khammanit et al., 2017; Moongkarndi et al., 2010). Thus,
consumption of vegetables and fruits has attracted growing interest
because of their significant role in attenuation of oxidative stress-
induced pathogenesis of diseases. Hence, the rationale of strategies
utilizes the natural antioxidants as therapeutic agents against oxida-
tive stress-induced diseases.

J Food Biochem. 2018;42:e12511. wileyonlinelibrary.com/journal/jfbc V

C 2018 Wiley Periodicals, Inc. | 1 of 11
https://doi.org/10.1111/jfbc.12511
2 of 11 | JAISUPA
Mangosteen (Garcinia mangostana L.) is a tropical plant belonging
to the family Guttiferae (also known as Clusiaceae). Its fruit peel con-
tains several phytochemicals including phenolic compounds that pri-
marily exhibit the antioxidant properties and has been used as a
traditional medicine for treating several illnesses (Kosem, Han, &
Moongkarndi, 2007). In addition, mangosteen peel extract (ME) has
been reported to exhibit antimicrobial, antiproliferation, and anticancer
properties (Kosem et al., 2007). The major bioactive compounds found
in mangosteen peel are xanthones (nonpolar compounds), flavonoids,
and condensed tannins or proanthocyanidins (polar compounds) (Fu,
Loo, Chia, & Huang, 2007). Flavonoids and proanthocyanidins act as
antioxidant agents, and that they protect against oxidative damage and
have many health benefits (Yao et al., 2004). Previous studies have
been reported the activities of xanthones or other nonpolar constitu-
ents, whereas the activities of the polar constituents have rarely been
described. Thus, study of polar fractions from mangosteen peel has
been of interest. Polar fractions of mangosteen peel have been shown
to have antioxidant and cytoprotective effects (Moongkarndi et al.,
2010; Ngawhirunpat et al., 2010; Weecharangsan et al., 2006). For
instance, the nonpolar fraction of ME containing xanthones exhibited
cytotoxic activity (Xu et al., 2014), whereas the polar fraction of ME eli-
cited potent antioxidant activity (Moongkarndi et al., 2010, 2014;
Suthammarak et al., 2016). Administration of ME resulted in an
improvement of cognitive function in animal models of scopolamine-
induced dementia (Sattayasai et al., 2013), and exhibited a decrease in
oxidative stress in whole blood and red blood cells of healthy humans
(Suthammarak et al., 2016). Moreover, treatment with ME protected
neuronal cells from beta-amyloid-induced cytotoxicity (Moongkarndi
et al., 2010), and protected SK-N-SH cells from hydrogen peroxide
(H2O2)-induced oxidative stress (Sattayasai et al., 2013). Even though
antioxidant activities of ME have been reported, the effects of phyto-
chemicals contained in ME on the production of antioxidant enzymes,
including its activity in the cells have not been fully defined. This study
aimed to evaluate the effects of ME on the inhibition of H2O2-induced
intracellular ROS production and on the upregulation of antioxidant
enzymes, including the mRNA expression and enzyme activity, in SK-
N-SH and HEK-293 cells. In addition, anti-lipid peroxidation, H2O2
scavenging activity, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scaveng-
ing activity assays were used to evaluate the antioxidant capacity of
ME and isolated compounds from ME.
2 | MATERIALS AND METHODS
2.1 | ME preparation
ME was obtained according to the method as previously described
(Moongkarndi et al., 2010, 2014). Briefly, powdered mangosteen peel
was macerated in ethanol with low heating, and the extracted solution
was dried under reduced-pressure using a rotary evaporator. The
obtained gummy syrup was then partitioned with ethyl acetate (EtOAc)
and water to fractionate the nonpolar fraction and the polar (water-solu-
ble) fraction (ME) from each other. Cytotoxic xanthones, such as
a-mangostin, were soluble in EtOAc and separated from water-soluble
ET AL.
contents in this step. The collected water-soluble fraction was spray-
dried to yield the powder extract (ME). The fingerprint of the ME was
determined by reversed-phase high performance liquid chromatography
(HPLC) (BDS HYPERSIL C18 from Thermo scientific, length 150 mm, I.D.
4.6 mm, UV detector 254 nm) with a gradient mobile phase system con-
sisting of 0.1% vol/vol formic acid and methanol at a flow rate of 1 mL/
min. a-Mangostin residue was analyzed by TLC developed in a mobile
phase consisting of EtOAc-hexane-ethanol (1:0.6:0.07, vol/vol/vol).
2.2 | Quantification of total phenolic and flavonoid
contents
Total phenolic and flavonoid contents were assayed by the Folin–Cio-
calteau and aluminum chloride colorimetric methods as previously
described with some modifications (Anantachoke et al., 2016). The
total phenolic contents in ME were calculated using a calibration curve
from gallic acid and expressed as mg gallic acid equivalent (GAE) per g
dry weight (DW). Total flavonoid contents of ME were calculated using
a calibration curve from quercetin and expressed as mg quercetin
equivalent (QE) per g DW. Assays were run in triplicate.
2.3 | DPPH scavenging assay
The DPPH scavenging activity was performed as previously described
(Moongkarndi et al., 2014). The scavenging activity is based on the abil-
ity of the extract to convert DPPH radical (purple) into DPPHH mole-
cule (yellow). Briefly, DPPH solution was sprayed onto the developed
TLC plate of ME to primarily screen for the antioxidant activity of each
separated bands. For the quantitative assay, 100 lL of extract solutions
in MeOH were mixed with 100 lL of 0.4 mM of DPPH in MeOH. The
reactions were incubated for 30 min in the dark at room temperature.
The absorbance was spectrophotometrically measured at 517 nm.
Ascorbic acid was used as the positive control. The scavenging activity
was calculated using the following equation:
Scavenging activity ð%Þ 5 ½12fðA1 2blankÞ=A0 2blankÞg 3 100;
where A1 was the absorbance of the mixture in the presence of the
extracts, A0 was the absorbance of the mixture without extract, and
the blank was the absorbance of MeOH.
2.4 | Cell culture
Human neuroblastoma (SK-N-SH) and human embryonic kidney-293
(HEK-293) cells were grown in Dulbecco’s Modified Eagle’s medium
(DMEM) supplemented with 10% fetal calf serum and 1% (vol/vol) of
penicillin/streptomycin solution. The cells were incubated at 378C in a
humidified 95% air with 5% CO2 and passaged routinely by trypsiniza-
tion as previously described (Khammanit et al., 2017).
2.5 | 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT) assay
Cell viability using MTT assay was applied in order to investigate an
appropriate concentration of ME as previously described (Anantachoke
JAISUPA ET AL. | 3 of 11

TA BL E 1 The specific primers for the human antioxidant genes

Genes specific primer Sense Antisense

0 0
CAT 5 -GCAGATACCTGTGAACTGTC-3 50 -GTAGAATGTCCGCACCTGAG-30

Mn-SOD 50 -GCACATTAACGCGCAGATCA-30 50 -AGCCTCCAGCAACTCTCCTT-30

HO-1 50 -CAGGCAGAGAGAATGCTGAG-30 50 -GCTTCACATAGCGCTGCA-30

GRe 50 -CAGTGGGACTCACGGAAGAT-30 50 -TTCACTGCAACAGCAAAACC-30

GAPDH 50 -CGAGATCCCTCCAAAATCAA-30 50 -GTCTTCTGGGTGGCAGTGAT-30

CAT 5 catalase; GAPDH 5 glyceraldehyde-3-phosphate dehydrogenase; GRe 5 glutathione reductase; HO-1 5 heme oxygenase-1; Mn-
SOD 5 manganese superoxide dismutase.

et al., 2016). Briefly, an amount of 2 3 104 cells were seeded into 96- KAPA SYBR FAST one-step RT-qPCR Kit (KAPA biosystems) and the
well plate and incubated overnight. Various concentrations of ME were Mx3005 RT PCR system (Stratagene) according to the manufacturer
added into each well to give the final concentration of 400, 200, 100, instructions with the specific primers (Table 1). The data were obtained
and 50 lg/mL. The experiments were performed in triplicate. The cells from three independent experiments.
were further incubated for 24 hr and then followed by 100 lL of 1%
wt/vol of MTT solution in serum-free DMEM for 3 hr. The intracellular
formazan crystal formed in living cells was dissolved with 100 lL of 2.8 | Determination of CAT activity
dimethyl sulfoxide (DMSO). The absorbance value of the solution at
CAT activity was examined in both SK-N-SH and HEK-293 cells by
570 nm directly represents relative cell numbers. Cell viability (%) was
using CAT activity assay kit (colorimetric) (Abcam). Briefly, the cells
expressed as a percentage relative to the vehicle-treated (control)
were seeded in a six-well plate (1 3 106 cells/wells) overnight and then
group.
treated with 100 lg/mL of ME for 24 hr. The enzyme activity was cal-
culated as mentioned in the manufacturer’s instructions. The CAT
2.6 | Determination of intracellular ROS levels in
activity was expressed as pmol/min/mL.
SK-N-SH cell
Intracellular ROS levels were determined as DCF fluorescent intensity
and performed as described previously (Moongkarndi et al., 2014) with 2.9 | Anti-lipid peroxidation in linoleic acid system
a slight modification. Briefly, 2 3 10 cells of SK-N-SH were seeded
4
Determination of anti-lipid peroxidation was assessed by using linoleic
into 96-well plate and incubated for 24 hr. Cells were further treated
acid system as previously described (Kosem et al., 2007). Briefly, to pre-
with 100 lL of ME (100 lg/mL) for 24 hr and then washed with phos-
pare linoleic acid mixture solution, 0.41 mL of 2.51% vol/vol linoleic
phate buffer saline (PBS, pH 7.4). The cells were further incubated with
acid in absolute ethanol (EtOH), 0.4 mL of each extract solution in
20 ,70 -dichlorfluorescein-diacetate (DCFH-DA) at final concentration 50
absolute EtOH (10 and 100 lg/mL final concentration), 0.8 mL of
lM for 45 min and washed out with PBS. To measure the intracellular
0.05 M phosphate buffer pH 7, and 0.39 mL of distilled water were
ROS level, 100 lL of 30 lM H2O2 in PBS was added. After 30 min of
mixed together. The mixtures were incubated at 378C in dark, and
incubation, the intracellular level of DCF was measured by fluorometer
determined for lipid peroxide formation within 10-day duration. To
with the excitation and emission wavelengths at 485 and 530 nm,
measure lipid peroxide formation, 30 lL of the mixtures were mixed
respectively. The controls were the cells treated and untreated with
with 140 lL of 75% EtOH, 30 lL of 0.02 M FeSO4 in 0.2 M HCl, and
H2O2. Vitamin E at 200 lg/mL was used as a positive control. The data
were obtained from three independent experiments. 30 lL of 30% KSCN. The mixtures were thoroughly mixed for 3 min
and then the absorbance was measured at 500 nm using spectropho-
tometer. a-Tocopherol (vitamin E) was used as the positive control.
2.7 | Analysis of antioxidant genes expression
Higher absorbance indicated higher lipid radical formation. The princi-
Gene expression was performed by using SK-N-SH and HEK-293 cells. ple of this assay was due to the oxidation of Fe21 to Fe31 by lipid radi-
The procedures were performed according to the previous study cal, resulting in the formation of Fe31-thiocyanate complex that
(Vongsak, Mangmool, & Gritsanapan, 2015). mRNA expressions of four provided deep red color. The inhibition of lipid peroxidation was calcu-
antioxidant genes, including CAT, manganese superoxide dismutase lated by the following equation:
(Mn-SOD), heme oxygenase-1 (HO-1), and glutathione reductase
Anti2lipid peroxidation activity ð%Þ 5 ½12fðA1 2blankÞ=
(GRe), were evaluated. Briefly, cells (1 3 106 cells/well) were seeded
ðA0 2blankÞg 3 100;
overnight in a six-well plate and further incubated with the extracts at
the concentration of 100 lg/mL for 8 hr in supplements-free DMEM. where A1 is the absorbance in the presence of extracts, A0 is the
The mRNAs were extracted using the GenJET RNA purification kit absorbance in the absence of extracts, and blank is the absorbance of
(Thermo Scientific). Quantitative mRNA levels were performed using mixture that contained PBS instead of linoleic acid.
4 of 11 | JAISUPA
2.10 | H2O2 scavenging assay
H2O2 scavenging activity assay was performed as previously described
(Jayaprakasha, Jaganmohan, & Sakariah, 2004) with a slight modification.
Briefly, 1 mL of various concentrations of extracts was added into 2 mL
of 20 mM H2O2 in 0.1 M PBS (pH 7.4), and then incubated for 10 min.
The absorbance of the reaction mixture was measured at 230 nm. The
H2O2 scavenging activity was calculated using the following equation;


Scavenging activity ð%Þ5 12fAsample 2blank =ðAcontrol 2blankÞg 3 100;
where Asample is the absorbance in the presence of extracts, Acontrol is
the absorbance of the control, and the blank was the absorbance in the
absence of H2O2.
2.11 | TLC analysis and structure elucidation of
individual compounds in ME
Thin layer chromatography (TLC) method was used to determine the
active constituents of ME as previously described (Majaw & Nongbet,
2013). Briefly, ME was subjected onto TLC plates (60 F254), and devel-
oped in mobile phase (EtOAc-hexane-EtOH; 1:0.6:0.07). The isolated
compounds were visualized under UV light and qualitatively screened
for phenolic compounds by spraying with 1% ferric chloride (FeCl3)
(Mabinya, Mafunga, & Brand, 2006). These phenolic compounds were
scraped from the TLC plate and recovered by extraction with MeOH.
The structures of these compounds were further elucidated by spectro-
scopic methods (NMR, IR, and MS). Vanillin in acidic condition was also
applied for testing of flavans structure of the compounds.
2.12 | Statistical analysis
To verify the statistical analysis, p values were obtained from Mann–
Whitney U test using software SPSS version 19. The data were presented
F I G U R E 1 HPLC chromatograms of ME illustrating the predominant; compound 1 [unknown, RT 17.8 min, compound 2 [proanthocyanidin
dimers, RT 40.3 min, and compound 3 [(2)-epicatechin, RT 47.8 min]
ET AL.
as mean 6 standard error of the mean (SEM) of three independent experi-
ments. p Value less than .05 was considered statistically significant.
3 | RESULTS
3.1 | ME preparation
After spray drying, ME was obtained as a bulky light brown powder. The
HPLC chromatogram revealed the fingerprint with three predominant
peaks eluted at the retention times 17.8 min (compound 1), 40.3 min
(compound 2), and 47.8 min (compound 3) (Figure 1). Xanthones, including
a-mangosteen were not found in ME as shown in the TLC chromatogram
(Figure 2). The yields of compounds 1, 2, and 3 were obtained approxi-
mately 0.08, 0.3, and 0.5% based on DW of the ME powder, respectively.
3.2 | Total phenolic and flavonoid contents of ME
Total phenolic content was estimated using the Folin–Ciocalteu method
which relied on the transfer of electrons from phenolic compounds to
the Folin–Ciocalteu reagent. We found that total phenolic content of ME
was approximately 62.15 6 3.57 mg GAE/g DW. In addition, the flavo-
noid content of ME was approximately 260.44 6 3.03 mg QE/g DW.
3.3 | DPPH scavenging activity of ME
We determined the antioxidant capacity of ME using a DPPH scaveng-
ing assay. This assay based on the measurement of the antioxidant abil-
ity to scavenge the DPPH radical and used vitamin C as the positive
control. Formation of a yellow color after spraying DPPH solution onto
the developed TLC plate indicated the antioxidant capacity of com-
pounds 1, 2, and 3 (Figure 3, lane D). For the quantitative assay, the
DPPH scavenging activities were expressed as the half maximal inhibi-
tory concentration (IC50) as shown in Table 2.
JAISUPA ET AL. | 5 of 11

FIGURE 3 TLC chromatogram of ME in different condition;

visualized under UV 254 nm (a), FeCl3-treated (b), reaction with
vanillin in acidic condition (c), reaction with DPPH solution (d)

found to exhibit the highest H2O2 scavenging activity (13.14 6 0.55

lg/mL). The IC50 values for the H2O2 scavenging activity of ME, com-
pound 1, and compound 2 were 22.15 6 2.75, 36.81 6 1.93, and
20.03 6 0.49, respectively (Table 2).

3.5 | Anti-lipid peroxidation activity of ME

FIGURE 2 TLC chromatogram developed in mobile phase system
consisting of ethyl acetate-hexane-ethanol (1:0.6:0.07) to compare Anti-lipid peroxidation activity of ME was determined by the ferric thi-
the Rf values of the isolated compounds of ME, standard
ocyanate method in a linoleic acid system. With this method peroxide
a-mangostin (a-MG) and standard xanthone (Xan)
formation occurred during the oxidation of linoleic acid. We monitored
Compound 1 that isolated from ME exhibited the lowest DPPH the levels of lipid peroxide formation for 10 days, and found that the
scavenging activity (41.57 6 2.89 mg/mL) compared with ME and other
compounds (Table 2). In addition, the DPPH scavenging activity of vita- T AB LE 2 The IC50 of H2O2 scavenging activity and DPPH scaveng-
min C (9.32 6 0.86 lg/mL) was higher than that of all compounds iso- ing activity of ME and vitamin C (mean 6 SEM)
lated from ME, whereas ME showed moderate DPPH scavenging H2O2 scavenging DPPH scavenging
activity (26.75 6 3.07 lg/mL) (Table 2). Compounds activity activity

ME 22.15 6 2.75 lg/mL 26.75 6 3.07 lg/mL

1 36.81 6 1.93 lg/mL 41.57 6 2.89 lg/mL

3.4 | H2O2 scavenging activity of ME
2 20.03 6 0.49 lg/mL 11.45 6 0.23 lg/mL
We next determined the antioxidant capacity of ME using a H2O2
3 13.14 6 0.55 lg/mL 19.31 6 2.39 lg/mL
scavenging assay and used vitamin C as the positive control with the
Vitamin C 38.53 6 1.99 lg/mL 9.32 6 0.86 lg/mL
IC50 of 38.53 6 1.99 lg/mL. As shown in Table 2, the compound 3 was
6 of 11 | JAISUPA ET AL.

manner. At the 100 lg/mL concentration, ME exhibited approximately

36% inhibition of lipid peroxidation of linoleic acid. On the other hand,
at the same concentration, vitamin E showed 36% inhibition, suggest-
ing that ME has similar anti-lipid peroxidation activity compare with
vitamin E. Furthermore, compound 1 (100 lg/mL) exhibited the highest
anti-lipid peroxidation with 65% inhibition, followed by compound 3
(53% inhibition) and compound 2 (41% inhibition) (Table 3). The results
clearly showed that all compounds isolated from ME had more anti-
lipid peroxidation activity than vitamin E at the same concentration
(100 lg/mL) (Table 3).

3.6 | Cytotoxic effects of ME

FIGURE 4 The absorbance value measured at 500 nm from day 0
to day 10; * indicated statistically significant compared to
untreated group on day 10; * p < .05 versus the control
absorbance values were not significantly different between the groups
from day 0 to day 5 (Figure 4). At day 10, the absorbance value in the
control group was predominantly higher than that of the ME-, com-
pound 1-, compound 2-, compound 3-, and vitamin E-treated groups
(Figure 4 and Table 3). As shown in Table 3, ME and isolated compound
1, 2, and 3 exhibited anti-lipid peroxidation in a dose-dependent
We assessed the cytotoxicity of ME in both SK-N-SH and HEK-293 cells
using the MTT assay to determine the optimal concentrations of ME for
use in further experiments. We found that at the concentrations of 50–
100 mg/mL, ME was not toxic to both cell types due to cell viability was
at least 90% without significant morphological change. Cell viability
began to decrease at ME concentrations higher than 200 mg/mL in SK-
N-SH and HEK-293 cells (Figure 5a,b, respectively). The half maximal
cytotoxic concentration (CTC50) was approximately 400 and 300 mg/mL
for SK-N-SH and HEK-293 cells, respectively. Therefore, ME at the con-
centration of 100 mg/mL was selected and used for further studies.
3.7 | Treatment with ME inhibits H2O2-induced
intracellular ROS production
We next investigated whether ME inhibits H2O2-induced intracellular
ROS production by using a fluorescent probe, DCFH-DA. After

TA BL E 3 Anti-lipid peroxidation activity of ME and vitamin E

measured on day 10

Average OD p Value
500 (subtracted Approximate (compared
Compounds from blank) % inhibition to control)

Control 1.749 0 –

ME, 10 lg/mL 1.181 32 .029*

ME, 100 lg/mL 1.117 36 .042*

Compound 1, 10 lg/mL 0.857 51 .021*

Compound 1, 100 lg/mL 0.605 65 .013*

Compound 2, 10 lg/mL 1.070 39 .017*

Compound 2, 100 lg/mL 1.028 41 .015*

Compound 3, 10 lg/mL 1.377 21 .133

Compound 3, 100 lg/mL 0.816 53 .001*

Vitamin E, 100 lg/mL 1.299 26 .073

Vitamin E 200 lg/mL 1.122 36 .044*

FIGURE 5 Bar graphs represented the relation of concentration
*Indicated statically significant.
and cell viability; SK-N-SH (a) and HEK-293 cells (b)
JAISUPA ET AL.
FIGURE 6 The comparison of DCF fluorescent intensities in
SK-N-SH cell in four conditions; * indicated p value < .05
incubation with H2O2, the intracellular ROS levels robustly increased
compare with the control (vehicle) in SK-N-SH cells (Figure 6). Treatment
with ME at a concentration of 100 mg/mL significantly inhibited H2O2-
induced ROS production, which has effect similar to those of vitamin E (a
potent antioxidant) (Figure 6). These results suggested that ME inhibits
cellular oxidative stress by preventing the production of ROS in the cells.
3.8 | Treatment with ME increased the mRNA
expression of CAT and HO-1
Antioxidant enzymes, including GRe, CAT, Mn-SOD, and HO-1 play
important role in antioxidant signaling cascades in several cell types.
Treatment with ME significantly elevated the mRNA levels of CAT and
HO-1 in SK-N-SH and HEK-293 cells (Figure 7a,b, respectively). In
addition, GRe mRNA expression tended to increase in ME-treated cells.
F I G U R E 7 The ratio of antioxidant enzymes mRNA and GAPDH mRNA expression in SK-N-SH (a) and HEK-293 cells (b); * p < .05 versus
untreated group
| 7 of 11
However, treatment with ME had no effect on mRNA expression of
Mn-SOD in both cell types (Figure 7). Collectively, our results demon-
strated that ME plays an important role in inhibition of oxidative stress
by enhancing the synthesis of antioxidant enzymes.
3.9 | Effects of ME on CAT enzyme activity in HEK-
293 and SK-N-SH cells
As shown in Figure 7, treatment with ME significantly increased the
mRNA expression of CAT. We further investigated the antioxidant
effect of ME on CAT enzyme activity. Treatment with ME (100 lg/mL)
markedly increased cellular activity of CAT in SK-N-SH cells (Table 4).
In addition, treatment with ME tended to increase CAT activity in
HEK-293 cells. These results suggested that the effects of ME on the
enzymatic activity of CAT may be attributed to its antioxidant effects
in cells.
3.10 | TLC analysis and structure elucidation of active
compounds in ME
HPLC chromatogram at peaks 1, 2, and 3 (Figure 1) are corresponded
to compound 1, 2, and 3, respectively, on the developed TLC plate
(Figure 3, lane A). The ferric chloride test is used to determine the pres-
ence of phenol structures in the isolated compounds from ME. We
found that compound 1, 2, and 3 were given the positive results with
this test, suggesting these compounds are phenolic compounds (Figure
3, lane B). Compound 2 and compound 3 were used for further anal-
ysis of structural elucidation. The high-resolution electrospray ioni-
zation mass spectrometry (HRESIMS) data for compound 3 exhibited
an [M 1 Na]1 ion with m/z 313.0712 that corresponded to a
8 of 11 | JAISUPA ET AL.

TA BL E 4 Catalase activity (pmol/min/mL) in SK-N-SH and HEK-

293 cells (mean 6 SEM)

Cells Vehicle-treated ME-treated

SK-N-SH 0.098 6 0.032 0.478 6 0.111*

HEK-293 0.206 6 0.062 0.255 6 0.197

*
Indicated statically significant.

molecular formula of C15H14O6. Based on the spectroscopic data,

we found that compound 3 was determined as (2)-epicatechin (Fig-
ure 1). Our data are consistent with previous studies showing the
similar spectroscopic data for (2)-epicatechin (Mendoza-Wilson &
Glossman-Mitnik, 2006; Orisakeye & Olugbade, 2014; Shu-Hua, Da-
Gang, Yun-Bao, & Xiao-Dong, 2003). In addition, standard (2)-epica-
techin and standard (2)-catechin were used to confirm that com-
pound 3 is (2)-epicatechin by a TLC technique (Figure 8).
The HRESIMS data for compound 2 showed an [M 1 Na]1 ion
with m/z 601.1314 that corresponded to a molecular formula of
C30H26O12. Based on the spectroscopic data, we found that compound
2 was identified as a proanthocyanidin dimer, which is commonly found
in ME (Ngawhirunpat et al., 2010; Yoshimura et al., 2015). Our data are
consistent with previous studies showing the similar spectroscopic
data for proanthocyanidin dimer (Fu et al., 2007; Hoyos et al., 2015;
Orisakeye & Olugbade, 2014). Furthermore, vanillin in acidic conditions
was used to check for the existence of flavan structures (Price, Scoyoc,
& Butler, 1978). We found that compound 2 and compound 3 reacted
with this agent, leading to the formation of a pink adduct (Figure 3,
lane C). However, compound 1 did not react with this agent, indicating
that compound 1 did not contain a flavan structure (Figure 3, lane C).

4 | DISCUSSION

In this study, we discovered a novel mechanism of ME on the inhibition

of cellular oxidative stress. We demonstrated that ME has antioxidant
effects by scavenging of ROS and inducing the synthesis and activity of
antioxidant enzymes such as CAT and HO-1 in SK-N-SH and HEK-293
cells.
Phytochemicals found in vegetables and fruits could exert antioxi-
dant effects in several ways such as the non-enzymatic pathway by
donating electrons to ROS; chelating metal ions that promote free radi-
cal production; inhibiting free radical-derived enzymes; upregulating
antioxidant enzymes; and regenerating membrane-bound antioxidants
(Amic, Davidovic-Amic, Beslo, Rastija, & Lucic, 2007; Packer, Weber, &
Rimbach, 2001). Consumption of natural products has attracted grow-
ing interest because of their significant role on inhibition of oxidative FIGURE 8 The TLC chromatogram compared compound 3 with
stress. Previous studies have demonstrated that phenolic compounds standard (2)-epicatechin ((2)-EP), standard (2)-catechin ((2)-CAT)
developed in mobile phase consisting of chloroform-ethyl acetate-
from various sources are effective in reduction of ROS production
formic acid (5:4:1 vol/vol/vol) and visualized under UV 254 nm
(Anantachoke et al., 2016; Khammanit et al., 2017; Kosem et al., 2007),
suggesting their roles in the prevention and treatment of oxidative (Anantachoke et al., 2016; Khammanit et al., 2017). Thus, phenolic
stress-related diseases. Antioxidant activities assayed by several meth- compounds are the major contributor to the total antioxidant activities
ods showed strongly positive relationship with total phenolic contents in natural products. Mangosteen peel is a rich source of natural
JAISUPA ET AL.
antioxidants such as phenolic compounds and has been shown to pro-
tect the cells from oxidative damage (Kosem et al., 2007).
Our data are consistent with these previous studies (Ngawhirunpat
et al., 2010; Yoshimura et al., 2015) showing that epicatechin and
proanthocyanidin dimer were found in mangosteen peel. Previous stud-
ies indicated that epicatechin possessed a cytoprotective effect via
antioxidant activity (Cuevas et al., 2009; Martín, Fernandez-Millan,
Ramos, Bravo, & Goya, 2014). Epicatechin reacts with free radicals to
form the stable quinones or stable phenoxyl radicals (Amic et al., 2007).
In addition, epicatechin does not have a 4-carbonyl group, but it has
many hydroxyl groups in the structure that can scavenge free radicals.
Epicatechin can donate its own electron to ROS and form stable epica-
techin-o-quinone (Shay et al., 2015). Proanthocyanidins are oligomers
and polymers and are the most abundant type of pronathocyanidins in
plant kingdom, and are presented in many foods, including fruits (Gu
et al., 2003). Proanthocyanidins consisting only of (2)-epicatechin/
(1)-catechin units are called procaynidins, whereas prodelphinidins are
oligomers and polymers of (epi)gallocatechin (Gu et al., 2003). Proan-
thocyanidins show high scavenging activities against ROS such as
superoxide, hydroxyl radical, and DPPH radicals (Lu & Foo, 2000;
Omar, Mullen, & Crozier, 2011). Our present study demonstrated that
(2)-epicatechin (compound 3) and proanthocyanidin dimer (compound
2) were found in ME (Figure 1). Thus, the antioxidant effects of ME
could be due to the properties of (2)-epicatechin and proanthocyanidin
dimer. However, other phytochemical compounds apart from polyphe-
nols might be involved in the antioxidant activities of ME and should
be investigated in the further study.
In our present study, antioxidant activities of ME were evaluated
using DPPH free radical scavenging activity, H2O2 scavenging activity,
and anti-lipid peroxidation assays. DPPH assay was used to measure
the direct involvement of ME as a primary antioxidant. ME and its iso-
lated compounds, including proanthocyanidin dimer, and (2)-epicate-
chin, were found to exhibit free radical scavenging activities. In
addition, H2O2 scavenging activity and anti-lipid peroxidation assays
can be used to investigate non-enzymatic antioxidant effects in vitro,
because these two oxidants are naturally formed in the human body.
ME displayed the scavenging effect against these ROS, and therefore,
ME may be useful for the prevention of oxidative damage in cells and
tissues. These scavenging activities of ME could be due to the presence
of several phenolic compounds. For instance, phenolic compounds
donate electrons to H2O2 molecules and convert them to water mole-
cules for scavenging of H2O2 (Bendary, Francis, Ali, Sarwat, & El Hady,
2013). Moreover, phenolic compounds found in ME interacted and
bound to the peroxyl radical to form a stable product to get rid of lipid
peroxide (Koroleva et al., 2014). Interestingly, ME displayed anti-lipid
peroxidation activity, which had effects similar to those of vitamin E
(a potent antioxidant). These findings were consistent with a previous
study showing that a crude methanolic extract of mangosteen
peel exhibited anti-lipid peroxidation activity (Kosem et al., 2007;
Ngawhirunpat et al., 2010).
Although compound 3 (epicatechin) displayed approximately two-
fold higher IC50 than vitamin C on weight basis in DPPH scavenging
assay, its IC50 is only approximately 1.2-fold higher than vitamin C on
| 9 of 11
molar basis (52.9 lM for vitamin C and 66.58 lM for epicatechin). In
case of compound 2 (proanthocyanidin dimer), its IC50 in DPPH assay is
approximately 11.45 lg/mL, which was lower than that of vitamin C. If
this value is converted into molar unit (19.8 lM), compound 2 become
more potent (2.5-fold) than vitamin C. Thus, these two active com-
pounds found in ME exhibit the potent free radical scavenging activity.
Oxidative stress in the cells can cause DNA damage, lipid peroxida-
tion, and protein modification, resulting in cell senescence and damage.
Our present study reported antioxidant effect of ME by attenuating
H2O2-induced intracellular ROS production in SK-N-SH cells.
Cells treated with ME showed the lower DCF intensity, indicating
that lower amounts of intracellular ROS, including H2O2, were found in
these cells. This phenomenon may be due to poor diffusion of H2O2
into the cells or poor retention of H2O2 in the cells due to the defense
mechanisms. Therefore, the cells in the abovementioned groups lacked
the catalyst to convert DCFH to DCF. This finding may be explained
by several possibilities. The first possibility is the effect of some active
compounds in ME remaining in the cells that are able to scavenge
H2O2 directly. The second possibility is an ability of active compounds
in ME to trigger the resistance of the cell membrane to H2O2
(El-Rahman, Hammouda, & Fakeir, 1995). The third possibility is an
induction of the production and the activity of antioxidant enzymes, as
described in this study. Taken together, these data indicated an excel-
lent ability of antioxidant compounds in ME to scavenge free radicals
directly or through the promotion of cellular processes.
Several antioxidant enzymes including GPx, GRe, HO-1, CAT, Mn-
SOD, CuZn-SOD, and GST are the major components of antioxidant
signaling cascade in many cells and tissue. The measurements of
changes in endogenous antioxidant enzymes activity and production
are considered as a sensitive biomarker of the response to cellular oxi-
dative stress. In biological systems, CAT, which is naturally found in the
human body, catalyzes the conversion of H2O2 to water molecules.
HO-1 is one of the heme oxygenase isoforms responsible for protect-
ing cells from oxidative stress conditions, inflammation, or cell death. In
addition, HO-1 has been reported to have benefits in metabolic dis-
eases (Araujo, Zhang, & Yin, 2012; Son, Lee, Chung, & Pae, 2013).
Epicatechin and proanthocyanidin dimer have been reported to be
antioxidant agents and exhibited antioxidant effects in the cells and tis-
sues. For example, epicatechin regulates the redox/oxidative status
through the NF-E2-related factor (Nrf-2) pathway, which regulates
antioxidant genes expression (Chang, Cho, & Wang, 2014). Proantho-
cyanidins can enhance antioxidant gene expression in a murine model
(Long et al., 2016). In addition, proanthocyanidins-rich grape seed
extract reduced cisplatin-induced attenuation of activities of antioxi-
dant enzymes, including GST, SOD, and CAT (Long et al., 2016). Con-
sistent with these previous studies, our present data demonstrate that
ME, which contained epicatechin and proanthocyanidin dimer, sup-
pressed cellular oxidative stress through the upregulation of antioxi-
dant enzymes, including CAT and HO-1 in SK-N-SH and HEK-293
cells. Moreover, treatment with ME elicited cellular antioxidant activity
by increasing enzyme activity of CAT. Thus, epicatechin and proantho-
cyanidin dimer act as the major antioxidant compounds in ME. Interest-
ingly, compound 1 also found in ME and has antioxidant activity. Even
10 of 11 | JAISUPA
though compound 1 showed weak antioxidant activity, compared to
proanthocyanidin (compounds 2), and epicatechin (compound 3), fur-
ther study is required to elucidate the structure of compound 1. Data
from our study and others provided evidences that ME elicits antioxi-
dant effects in many cell types through an increase in the enzyme
activity and synthesis of antioxidant enzymes.
5 | CONCLUSIONS
Mangosteen peel is an interesting source of a number of antioxidant
compounds, including epicatechin and proanthocyanidin dimer. These
active compounds exhibited many benefits including excellent antioxi-
dant activity and cytoprotection. The antioxidant effects of ME have
been associated with both nonenzymatic pathways by directly scav-
enging ROS, and enzymatic pathways by enhancing the production and
activity of antioxidant enzymes, including CAT and HO-1. Finally, we
suggested that ME is a high potential natural product as a complemen-
tary or herbal medicine used for the treatment of oxidative stress-
related diseases.
AC KNOW LEDG MENT S
This study was supported by the grant from the Department for
Development of Thai Traditional and Alternative Medicine, Ministry
of Public Health. We would like to give special thanks to Professor
Dr. Neelobol Neungton for research suggestion and Assoc. Professor
Dr. Weena Jiratchariyakul for the discussion on the isolating and
structure elucidation of bioactive compounds. We would like to
thank Mrs. Angkana Jongsawadipatana for her assistances concern-
ing statistical calculations.
CONFLIC T OF I NTE R ES T
There was no conflict of interest concerning in this study.
ORCI D
Supachoke Mangmool http://orcid.org/0000-0003-3223-1551
RE FE RE NCE S
Amic, D., Davidovic-Amic, D., Beslo, D., Rastija, V., & Lucic, B. (2007).
Trinajstic N. SAR and QSAR of the antioxidant activity of flavonoids.
Current Medicinal Chemistry, 14, 827–845.
Anantachoke, N., Lomarat, P., Praserttirachai, W., Khammanit, R., & Man-
gmool, S. (2016). Thai fruits exhibit antioxidant activity and induction
of antioxidant enzymes in HEK-293 cells. Evidence-Based Complemen-
tary and Alternative Medicine, 2016, 6083136.
Apel, K., & Hirt, H. (2004). Reactive oxygen species: Metabolism, oxida-
tive stress, and signal transduction. Annual Review of Plant Biology,
55(1), 373–399.
Araujo, J. A., Zhang, M., & Yin, F. (2012). Heme oxygenase-1, oxidation,
inflammation, and atherosclerosis. Frontiers in Pharmacology, 3, 119.
Bendary, E., Francis, R. R., Ali, H. M. G., Sarwat, M. I., & El Hady, S.
(2013). Antioxidant and structure–activity relationships (SARs) of
some phenolic and anilines compounds. Annals of Agricultural Scien-
ces, 58, 173–181.
ET AL.
Chang, C. F., Cho, S., & Wang, J. (2014). (2)-Epicatechin protects hemor-
rhagic brain via synergistic Nrf2 pathways. The Annals of Clinical and
Translational Neurology, 1, 258–271.
Cuevas, E., Limon, D., Perez-Severiano, F., Diaz, A., Ortega, L., Zenteno,
E., & Guevar, A. J. (2009). Antioxidant effects of epicatechin on the
hippocampal toxicity caused by amyloid-beta 25–35 in rats. The Euro-
pean Journal of Pharmacology, 616, 122–127.
El-Rahman, A., Hammouda, M. A., & Fakeir, A. (1995). Flow Cytometric
evaluation of erythrocyte response to oxidant stress. Cytometry,
20(1), 19–22.
Fu, C., Loo, A. E. K., Chia, F. P. P., & Huang, D. (2007). Oligomeric proan-
thocyanidins from mangosteen pericarps. Journal of Agricultural and
Food Chemistry, 55, 7689–7694.
Gu, L., Kelm, M. A., Hammerstone, J. F., Beecher, G., Holden, J.,
Haytowitz, D., & Prior, R. L. (2003). Screening of foods containing
proanthocyanidins and their structural characterization using LC-MS/
MS and thiolytic degradation. Journal of Agricultural and Food Chemis-
try, 59, 1363–1369.
Hoyos, M. N., Sanchez-Patan, F., Masis, R. M., Martin-Alvarez, P. J., Ram-
irez, W. Z., Monagas, M. J., & Bartolome, B. (2015). Phenolic assess-
ment of Uncaria tomentosa L. (cat’s claw): Leaves, stem, bark and
wood extracts. Molecules, 20, 22703–22717.
Jayaprakasha, G. K., Jaganmohan, R. L., & Sakariah, K. K. (2004). Antioxi-
dant activities of flavidin in different in vitro model systems. Bioor-
ganic and Medicinal Chemistry, 12, 5141–5146.
Khammanit, R., Lomarat, P., Anantachoke, N., Sato, V. H., Ungsurungsie,
M., & Mangmool, S. (2017). Inhibition of oxidative stress through the
induction of antioxidant enzymes of pigmented rice bran in HEK-293
cells. Natural Product Communications, 12, 1107–1110.
Koroleva, O., Torkova, A., Nikolaev, I., Khrameeva, E., Fedorova, T.,
Tsentalovich, M., & Amarowicz, R. (2014). Evaluation of the antiradi-
cal properties of phenolic acids. International Journal of Molecular
Sciences, 15, 16351–16380.
Kosem, N., Han, Y. H., & Moongkarndi, P. (2007). Antioxidant and cyto-
protective activities of methanolic extract from Garcinia mangostana
hulls. Science Asia, 33, 283–292.
Long, M., Yang, S., Han, J., Li, P., Zhang, Y., Dong, S., . . . He, J. (2016).
The protective effect of grape-seed proanthocyanidin extract on oxi-
dative damage induced by zearalenone in Kunming mice liver. Inter-
national Journal of Molecular Sciences, 17(6), 808.
Lu, Y., & Foo, L. Y. (2000). Antioxidant and radical scavenging activ-
ities of polyphenols from apple pomace. Food Chemistry, 68(1),
81–85.
Mabinya, L. V., Mafunga, T., & Brand, J. M. (2006). Determination of
ferulic acid and related compounds by thin layer chromatography.
African Journal of Biotechnology, 5, 1271–1273.
Majaw, S., & Nongbet, A. (2013). Evaluation of antioxidant activity of
TLC fractions of Clerodendron colebrookianum leaf extract containing
flavonoids. International Journal of Pharmacy and Pharmaceutical Scien-
ces, 5, 90–94.
 Fern
Martín, M. A., andez-Mill
an, E., Ramos, S., Bravo, L., & Goya, L.
(2014). Cocoa flavonoid epicatechin protects pancreatic beta cell via-
bility and function against oxidative stress. Molecular Nutrition and
Food Research, 58, 447–456.
Mendoza-Wilson, A. M., & Glossman-Mitnik, D. (2006). Theoretical study
of the molecular properties and chemical reactivity of (1)-catechin
and (-)-epicatechin related to their antioxidant ability. Journal of
Molecular Structure, 761, 97–106.
Moongkarndi, P., Jaisupa, N., Samer, J., Kosem, N., Konlata, J., Rodpai, E.,
& Pongpan, N. (2014). Comparison of the biological activity of two
JAISUPA ET AL.
different isolates from mangosteen. Journal of Pharmacy and Pharma-
cology, 66, 1171–1179.
Moongkarndi, P., Srisawat, C., Saetun, P., Jantaravinid, J., Peerapittaya-
mongkol, C., Soi-Ampornkul, R., . . . Neungton, N. (2010). Protective
effect of mangosteen extract against beta-amyloid-induced cytotoxic-
ity, oxidative stress and altered proteome in SK-N-SH cells. Journal of
Proteome Research, 9, 2076–2086.
Ngawhirunpat, T., Opanasopi, P., Sukma, M., Sittisombut, C., Kat, A., &
Adachi, I. (2010). Antioxidant, free radical-scavenging activity and
cytotoxicity of different solvent extracts and their phenolic constitu-
ents from the fruit hull of mangosteen (Garcinia mangostana).
Pharmaceutical Biology, 48(1), 55–62.
Omar, M. H., Mullen, W., & Crozier, A. (2011). Identification of proantho-
cyanidin dimers and trimers, flavone C-glycosides, and antioxidants in
Ficus deltoidea, a Malaysian herbal tea. Journal of Agricultural and
Food Chemistry, 59, 1363–1369.
Orisakeye, O. T., & Olugbade, T. A. (2014). Epicatechin and procyanidin
B2 in the stem and root bark of Sterculia tragacantha Lindl (Sterculia-
ceae). Medicinal Chemistry, 4, 334–337.
Packer, L., Weber, S. U., & Rimbach, G. (2001). Molecular aspects of
alpha-tocotrienol antioxidant action and cell signalling. The Journal of
Nutrition, 131(2), 369S–373S.
Price, M. L., Scoyoc, S. V., & Butler, L. G. (1978). A critical evaluation of
the vanillin reaction as an assay for tannin in sorghum grain. Journal
of Agricultural and Food Chemistry, 26, 1214–1218.
Sattayasai, J., Chaonapan, P., Arkaravichie, T., Soi-Ampornkul, R., Junnu,
S., Charoensilp, P., . . . Moongkarndi, P. (2013). Protective effects of
mangosteen extract on H2O2-induced cytotoxicity in SK-N-SH cells
and scopolamine-induced memory impairment in mice. PLoS One,
8(12), e85053.
€ttemann, M.
Shay, J., Elbaz, H. A., Lee, I., Zielske, S. P., Malek, M. H., & Hu
(2015). Molecular mechanisms and therapeutic effects of (2)-epicate-
chin and other polyphenols in cancer, inflammation, diabetes, and neu-
rodegeneration. Oxidative Medicine and Cellular Longevity, 2015, 1.
Shu-Hua, Q., Da-Gang, W., Yun-Bao, M., & Xiao-Dong, L. (2003). A
novel flavane from Carapa guianensis. Acta Botanica Sinica, 45,
1129–1133.
View publication stats
| 11 of 11
Son, Y., Lee, J. H., Chung, H., & Pae, H. (2013). Therapeutic roles of
heme oxygenase-1 in metabolic disease: Curcumin and resveratrol
analogues as possible inducers of heme oxygenase-1. Oxidative Medi-
cine and Cellular Longevity, 2013, 639541.
Suthammarak, W., Numpraphrut, P., Charoensakdi, R., Neungton, N.,
Tunrungruangtavee, V., Jaisupa, N., . . . Muangpaisan, W. (2016). Anti-
oxidant enhancing property of the polar fraction of mangosteen peri-
carp extract and evaluation of its safety in human. Oxidative Medicine
and Cellular Longevity, 2016, 1293036.
Vongsak, B., Mangmool, S., & Gritsanapan, W. (2015). Antioxidant activ-
ity and induction of mRNA expressions of antioxidant enzymes in
HEK-293 cells of Moringa oleifera leaf extract. Planta Medicine, 81,
1084–1089.
Weecharangsan, W., Opanasopit, P., Sukma, M., Ngawhirunpat, T.,
Sotanaphun, U., & Siripong, P. (2006). Antioxidative and
neuroprotective activities of extracts from the fruit hull of mangos-
teen (Garcinia mangostana Linn.). Medical Principles and Practice, 15,
281–287.
Xu, Z., Huang, L., Chen, X. H., Zhu, X. F., Qian, X. J., Feng, G. K., . . . Li,
H. J. (2014). Cytotoxic prenylated xanthones from the pericarps of
Garcinia mangostana. Molecules, 19(2), 1820–1827.
Yao, L. H., Jiang, Y. M., Shi, J., Tomas-Barberan, F. A., Datta, N., Singanu-
song, R., & Chen, S. S. (2004). Flavonoids in food and their health
benefits. Plant Foods for Human Nutrition, 59, 113–122.
Yoshimura, M., Ninomiya, K., Tagashira, Y., Maejima, K., Yoshida, T., &
Amakura, Y. (2015). Polyphenolic constituents of the pericarp of
mangosteen (Garcinia mangostana L.). Journal of Agricultural and Food
Chemistry, 63, 7670–7674.
How to cite this article: Jaisupa N, Moongkarndi P, Lomarat P,
et al. Mangosteen peel extract exhibits cellular antioxidant activ-
ity by induction of catalase and heme oxygenase-1 mRNA
expression. J Food Biochem. 2018;42:e12511. https://doi.org/
10.1111/jfbc.12511