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Petroselinum crispum has antioxidant properties, protects against DNA damage and inhibits proliferation and migration of cancer cells


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Background Petroselinum crispum (English parsley) is a common herb of the Apiaceae family that is cultivated throughout the world and is widely used as a seasoning condiment. Studies have shown its potential as a medicinal herb. In this study, P. crispum leaf and stem extracts were evaluated for their antioxidant properties, protection against DNA damage in normal 3T3-L1 cells, and the inhibition of proliferation and migration of the MCF-7 cells.ResultsThe dichloromethane extract of P. crispum exhibited the highest phenolic content (42.31 ± 0.50 mg GAE g−1) and ferric reducing ability (0.360 ± 0.009 mmol g−1) of the various extractions performed. The extract showed DPPH radical scavenging activity with an IC50 value of 3310.0 ± 80.5 µg mL−1. Mouse fibroblasts, 3T3-L1, pre-treated with 400 µg mL−1 of the extract showed 50.9% protection against H2O2-induced DNA damage, suggesting its potential in cancer prevention. The extract (300 µg mL−1) inhibited H2O2-induced MCF-7 cell migration by 41 ± 4%. As cell migration is necessary for metastasis of cancer cells, inhibition of migration is an indication of protection against metastasis.ConclusionP. crispum has health promoting properties with the potential to prevent oxidative stress-related diseases and can be developed into functional food.
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Research Article
Received: 13 November 2014 Accepted article published: 13 January 2015 Published online in Wiley Online Library:
( DOI 10.1002/jsfa.7078
Petroselinum crispum has antioxidant
properties, protects against DNA damage and
inhibits proliferation and migration of cancer
Esther Lai-Har Tang, Jayakumar Rajarajeswaran, ShinYee Fung and
MS Kanthimathi*
BACKGROUND: Petroselinumcrispum (English parsley) is a common herb of the Apiaceae family that is cultivated throughout the
world and is widely used as a seasoning condiment. Studies have shown itspotential as a medicinal herb. In this study, P. crispum
leaf and stem extracts were evaluated for their antioxidant properties, protection against DNA damage in normal 3T3-L1 cells,
and the inhibition of proliferation and migration of the MCF-7 cells.
RESULTS: The dichloromethane extract of P.crispum exhibited the highest phenolic content (42.31 ±0.50 mg GAE g1) and ferric
reducing ability (0.360 ±0.009 mmol g1) of the various extractions performed. The extract showed DPPH radical scavenging
activity with an IC50 value of 3310.0 ±80.5 𝛍gmL
1. Mouse fibroblasts (3T3-L1) pre-treated with 400 𝛍gmL
1of the extract
showed 50.9% protection against H2O2-induced DNA damage, suggesting its potential in cancer prevention. The extract (300 𝛍g
mL1) inhibited H2O2-induced MCF-7 cell migration by 41% ±4%. As cell migration is necessary for metastasis of cancer cells,
inhibition of migration is an indication of protection against metastasis.
CONCLUSION: Petroselinum crispum has health-promoting properties with the potential to prevent oxidative stress-related
diseases and can be developed into functional food.
© 2015 Society of Chemical Industry
Keyword s: Petroselinum crispum; antioxidant activity; antiproliferative activity ; DNA protection; MCF-7 cell migration; hydrogen peroxide
Petroselinum crispum (Mill) Nyman ex AW Hill, commonly known
as English parsley, is a culinary and medicinal herb of the Api-
aceae family that grows up to 30 – 100 cm high.1The herb has
been used to flavor the cuisines of South East Asia, China, India,
South America and Mexico.2Although native to Europe and west-
ern Asia, the herb is now cultivated and consumed throughout the
world.3The leaves and stems, either fresh or dried, as well as the
seeds, have been employed in the food, pharmaceutical and cos-
metic industries.4In folk medicine, the aerial part of P. crispum is
used to treat hemorrhoids, the stem for urethral inflammation, and
the root is used to pass kidney stones5and improve brain func-
tion and memory.6Additionally, P. crispum is used as a carmina-
tive, stomachic, emmenagogic, abortifacient and nutritive agent.7
Studies have shown that P. crispum has hypoglycemic, diuretic,
hypolipidemic, antimicrobial, anticoagulant and hepatoprotective
The chemical composition and pharmacological properties
of P. crispum have been previously reported in various studies.
The herb contains flavonol glycosides of quercetin, apiol, myris-
ticin and luteolin. Terpenes, phthalides, furanocoumarins, apiin,
carotenoids, ascorbic acid and tocopherol are also present in
P. crispum.9,10 Supplementation of diets with fresh P. crispum
leaves increases the antioxidant capacity of plasma in rats11 and
decreases oxidative stress in humans.12 Zheng et al.13 reported the
inhibition of benzo[a]pyrene-induced tumorigenesis in the lungs
of mice by myristicin, a major volatile aromatic constituent of
parsley leaf oil. The vast health-promoting properties associated
with P. crispum warrant further study. Previous investigations
on P. crispum mostly focused on its antioxidant properties.14
As phenolic compounds and antioxidant activities depend on
variety, location and growth conditions of the plant, data on the
antioxidant activity of P. crispum are still relevant and useful. The
effect of P. crispum leaves and stems on the two most common
cancers in humans breast cancer and colon cancer are unclear
and lacking thus far. The main aim of this work was to investigate
the antioxidant activities and protection against DNA damage by
Correspondence to: MS Kanthimathi, Centre for NaturalProducts and Drug D is-
covery (CENAR) and Department of Molecular Medicine, Faculty of Medicine,
University of Malaya, 50603 Kuala Lumpur, Malaysia.
Centre for Natural Products and Drug Discovery (CENAR) and Department of
Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala
Lumpur, Malaysia
J Sci Food Agric (2015) © 2015 Society of Chemical Industry EL-H Tang et al.
extracts of P. crispum leaves and stems and their inhibition of the
proliferation and H2O2-induced migration of the breast cancer cell
line MCF-7. To the best of our knowledge, this is the first study
reporting on the effects of P. crispum on DNA protection and
inhibition of MCF-7 cell migration.
Materials and methods
Analytical-grade solvents were purchased from Fisher Scientific
(Loughborough, UK). Dimethyl sulfoxide (DMSO) and H2O2were
purchased from Univar (Ingleburn, NSW, Australia). Chemicals,
polyphenolic standards (gallic acid, quercetin, rutin), proteinase
K and RPMI-1640 were obtained from Sigma-Aldrich (St Louis,
MO, USA). Dulbecco’s Modified Eagle Medium (DMEM) was
purchased from Lonza (Basel, Switzerland) and fetal bovine
serum (FBS) was obtained from iDNA Biotechnology, Singapore.
3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT), Folin– Ciocalteu phenol reagent, tris(hydroxymethyl)
aminomethane and ethidium bromide were purchased from
Merck (Darmstadt, Germany). Sodium dodecyl sulfate (SDS) was
from Bio-Rad (Hercules, CA, USA). TRIzol®reagent was purchased
from Life Technologies (Carlsbad, CA, USA). RNase A and DNA
ladders were obtained from Thermo Scientific (Carlsbad, CA,
USA). Ultrapure water from a Milli-Q-plus filter system (Millipore,
Billerica, MA, USA) was used throughout the experiments.
Cell lines and cell culture conditions
Two human breast adenocarcinoma cell lines – MCF-7 and
MDA-MB-231 – and the human colorectal adenocarcinoma cell
line HT-29 were used in the antiproliferative study. The MCF-7
cell line expresses the estrogen receptor whereas MDA-MB-231
cells do not. The study on the inhibition of cell migration, trypan
blue dye exclusion assay and DNA fragmentation analysis was per-
formed using MCF-7 cells. Mouse fibroblasts (3 T3-L1) were used in
the comet assay to estimate protection against H2O2-induced DNA
damage. All cells were purchased from the American Type Culture
Collection (Manassas, VA, USA). MCF-7 and HT-29 cancer cells
were routinely cultured in RPMI-1640. MDA-MB-231 and 3T3-L1
were grown in DMEM. The cells were supplemented with 10%
fetal bovine serum (FBS), 100 U mL1penicillin and 100 μgmL
streptomycin. Cells were grown at 37 C in a humidified incubator
with 5% CO2.
Plant material
Fresh P. crispum leaves and stems were purchased from the
local market in Kuala Lumpur, Malaysia. The plant was identi-
fied by Dr M Sugumaran, Institute of Biological Sciences, Uni-
versity of Malaya, and the voucher specimen (KLU47745) was
deposited in the University of Malaya herbarium. The leaves and
stems were washed under running tap water and finally rinsed
with distilled water. The plant parts were then freeze dried,
weighed, ground into fine powder and stored at 20 Cuntil
Preparation of P. crispum extracts
Powdered leaves and stems of P. crispum were extracted through
sequential extraction using hexane, dichloromethane, ethyl
acetate, methanol and water. Briefly, 120 g powdered leaves and
stems were extracted in 600 mL hexane (1:5 w/v) for 6 h at 40 Con
a hotplate stirrer. Extracts were then filtered through Whatman no.
1 filter paper and the resulting residue was re-extracted twice with
fresh hexane. The remaining residue was subsequently extracted
three times each with dichloromethane, followed by ethyl acetate,
methanol and water. Each filtrate (except for the aqueous extract,
which was concentrated to dryness in a freeze-dryer) was con-
centrated to dryness under reduced pressure at 40 Cusinga
rotary evaporator. The dried extracts were stored at 20 C. For
bioassays, the dried extracts were dissolved in DMSO and diluted
in ultrapure water to make appropriate extract concentrations.
The final concentration of DMSO in reaction mixtures was less
than 1%. All dissolved extracts were kept at 4 C throughout the
Determination of total phenolic content
The total phenolic content of P. crispum extracts was determined
using the Folin-Ciocalteau method,15 with some modifications.
Briefly, 500 μL of 1:10 Folin– Ciocalteau phenol reagent was added
to 1 0 μL of sample (dissolved in 10% DMSO), standard or positive
control. The mixture was mixed and allowed to stand for 5min
before the addition of 350 μL of 10% sodium carbonate. The
resulting reaction mixture was incubated in the dark at room
temperature for a further 2 h. Absorbance was then measured at
765 nm using a spectrophotometer. Gallic acid (50– 500 mg L1in
10% DMSO) was used as the standard. Rutin and quercetin were
used as positive controls. Results were expressed in milligrams
of gallic acid equivalents (GAE) per gram of dried extract. All
experiments were carried out in triplicate.
Ferric reducing antioxidant power (FRAP) assay
Ferric reducing activity of P. crispum extracts was estimated
based on the assay by Benzie and Strain16 with slight mod-
ifications. A working reagent was prepared fresh by mixing
10 mL of 300 mmol L1acetate buer with 1mL of 10mmolL
2,4,6-tripyridyl-s-triazine (TPTZ) in 40 mmol L1hydrochloric acid
and 1 mL of 20 mmol L1ferric chloride hexahydrate (FeCl3.6H2O).
The freshly prepared FRAP reagent was pre-warmed at 37Cfor
5 min, after which a blank reading was taken at 595 nm using
a plate reader. Subsequently, 3 μL sample (dissolved in 10%
DMSO), standard or positive control and 9 μL water were added to
90 μL of the FRAP reagent. Absorbance readings were measured
instantly upon addition of the FRAP reagent and again at 4 min
after the start of the reaction. The change in absorbance in the
4 min reaction was calculated by comparison with a FeSO4.7H2O
standard cur ve (100 –1000 μmol L1) tested in parallel. Rutin
and quercetin were used as positive controls. Results were
expressed as millimoles of ferric reducing activity of the extracts
per gram of dried extract. All experiments were carried out
in triplicate.
1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging
The radical scavenging activity of P. crispum extracts was deter-
mined by the DPPH radical scavenging assay,17 with some modi-
fications. Petroselinum crispum extract (20 μL) was added to 120 μL
of 0.04 mg mL1DPPH solution in methanol. The extracts tested
ranged from 0 to 5000 μgmL
1(dissolved in 10% DMSO). The solu-
tions were mixed well and incubated in the dark for 30min. The
reduction of DPPH absorption was measured at 515 nm using a
plate reader. Rutin and quercetin were used as positive controls.
All determinations were performed in triplicate. The DPPH radi-
cal scavenging activity was calculated according to the following © 2015 Society of Chemical Industry J Sci Food Agric (2015)
Antioxidant, DNA-protective and anticancer properties of Petroselinum crispum
Percentage inhibition =(Absorbancecontrol Absorbancesample)
Absorbancecontrol ×100
The results were expressed as half-maximal inhibitory concen-
tration (IC50), i.e. the concentration of the plant extract required to
scavenge 50% of the total DPPH radicals available.
Inhibition of proliferation (MTT assay)
The antiproliferative activity of P. crispum extracts on MCF-7,
MDA-MB-231 and HT-29 cancer cell lines was estimated using
the MTT assay as described by Mosmann.18 Briefly, cells supple-
mented with 5% FBS were seeded (5 ×103cells per well) in 96-well
plates and were allowed to grow at 37 C in a humidified atmo-
sphere with 5% CO2. After 24 h incubation, the cells were treated
with different concentrations of extract (0500 μgmL
1) for a fur-
ther 48 h. Vehicle-control wells with cells only and diluent-control
wells with similar DMSO concentrations as in the treatment were
included. After incubation, 10 μL of 5 mg mL1MTT bromide in
phosphate-buffered saline (PBS) were added to each well. The
plates were reincubated for 4h, after which media and MTT were
removed by aspiration. DMSO (100 μL) was added to each well
to dissolve the formazan crystals. Absorbance was read using a
microtiter plate reader at 595 nm. All measurements were per-
formed in triplicate. The percentage inhibition of cell proliferation
was calculated using the following formula:
Percentage inhibition =(Absorbancecontrol Absorbancetreated)
Absorbancecontrol ×100
Comet assay
The DNA protective effect of P. crispum was estimated using the
comet assay.19 Mouse fibroblasts (3 T3-L1) were cultured in 12-well
culture plates (1 ×105cells per well) for 24 h. The cells were then
pre-treated with dichloromethane extracts of P. crispum, at con-
centrations of 100 –400 μgmL
1for a further 24 h. The control con-
tained DMSO instead of extract. After pre-treatment, cells were
treated with 100 μmol L1of H2O2(final concentration in the well)
for 60 min on ice to induce DNA damage. Cells were then harvested
using a cell scraper, centrifuged and resuspended in 1mL PBS. The
cell suspension (25 μL) was mixed with 75 μL of 0.6% low-melting
agarose and the suspension was spread on a frosted microscopic
slide pre-coated with 250 μL of 0.8% normal melting agarose, cov-
ered with a cover slip, and then allowedto solidify on ice for 10 min.
The cover slips were removedand the slides were immersed in cold
lysis solution containing 1% SDS, 2.5 mol L1NaCl, 100 mmol L1
Na2ethylenediaminetetraacetic acid (Na2EDTA), 1% Triton X-100
and 10% DMSO (with the DMSO added just before use) for 1h
at 4 C in the dark. The slides were arranged in an electrophore-
sis tank filled with pre-chilled electrophoretic buffer (1 mmol L1
Na2EDTA and 300 mmol L1NaOH) and incubated for 20 min. Elec-
trophoresis was conducted in the same buffer at 25 V (300mA) for
20 min. The slides were washed with 0.4 mol L1Tris –HCl (pH 7.5)
and stained with 20 μgmL
1ethidium bromide for viewing under
a fluorescence microscope. The comet tail length was measured
using an ocular micrometer. A total of 50 individual cells were
screened per slide.20 The assay was carried out in triplicate. Results
were expressed in percent DNA protection, calculated using the
following formula:
DNAprotection (%)=(taillengthcontrol taillengthtreatment )
taillengthcontrol ×100
Scratch motility assay
The inhibitory effect of P. crispum on MCF-7 cell migration was
tested using the scratch motility assay. MCF-7 cells (3.5×105cells
per well) were seeded in a 24-well plate and grown for 24h.
The confluent cell monolayer was then scratched vertically with a
pipette tip, washed twice with PBS and incubated with media con-
taining P. crispum dichloromethane extract (0, 200, 300, 400, 500
and 600 μgmL
a final concentration of 1 μmol L1in the cell suspension to stim-
ulate the proliferation and migration of MCF-7 cells. The number
of cells in the denuded area were photographed and counted at 0
and 24 h incubation. The experiment was performed in triplicate.
The percentage inhibition was calculated as described by Sato and
Rifkin.21 Percentage inhibition =100 [(cell no. in denuded area of
sample / cell no. in denuded area of control)×100].
Trypan blue dye exclusion assay
A hemocytometer-based trypan blue dye exclusion cell quanti-
tation and viability assay was used to confirm the antiprolifera-
tive activity of P. crispum. MCF-7 cells were seeded in a six-well
plate (1.5 ×106cells per well) supplemented with 5% FBS and
allowed to grow at 37 C in a humidified atmosphere with 5%
CO2. After 24 h incubation, the cells were treated with differ-
ent concentrations of extract (0800 μgmL
1) for a further 48 h.
Diluent-control wells with similar DMSO concentrations as in the
treatment were included. Following treatment, the cells were col-
lected using 0.25% trypsinEDTA, pelleted and resuspended in
medium. The cells were then stained with an equal volume of
0.2% (w/v) trypan blue solution and the number of viable cells was
counted using a hemocytometer under an inverted microscope.
All measurements were performed in triplicate.
DNA fragmentation analysis
Agarose gel electrophoresis was used to investigate DNA fragmen-
tation in cells treated with P. crispum. MCF-7 cells (2 ×106mL1)
were grown in a 75 cm2culture flask for 24 h. The cells were
then treated with P. crispum dichloromethane extract (500 or
800 μgmL
1) for 24 or 48 h. After treatment, cells were lysed and
the DNA was extracted using TRIzol®reagent according to the
manufacturer’s protocol. Extracted DNA was treated with RNase
C for 1 h and proteinase K (200 μgmL
50 C for 2 h. The purified DNA was stored at 20 C until DNA
electrophoresis. The experiment was performed in triplicate. Iso-
lated DNA samples were subjected to electrophoresis in 1.8% (w/v)
agarose gel (in TAE buffer) impregnated with 0.5 μgmL
bromide and run at 90 V for 50 min. The gel was observed under
UV illumination and visualized using a gel documentation system
Statistical analysis
Data are presented as mean ±standard deviation (SD). Statistical
analysis was performed by one-way analysis of variance (ANOVA)
with Tukey’s multiple comparisons and Student’s t-test. A P-value
of <0.05 was considered statistically significant. Pearson’s correla-
tion coefficient was used to assess the correlation between TPC,
J Sci Food Agric (2015) © 2015 Society of Chemical Industry EL-H Tang et al.
Table 1. Phenolic content, ferric reducing antioxidant power and DPPH radical scavenging activity of Petroselinum crispum extracts
P. cri s p u m extract/
positive controls
Total phenolic content
(mg GAE g1)
FRAP value
(mmol g1)
DPPH radical scavenging
activity (μgmL
Hexane 20.17 ±1.35b 0.075 ±0.002b 4485.0 ±78.0b
Dichloromethane 42.31 ±0.50d 0.360 ±0.009d 3310.0 ±80.5a
Ethyl acetate 32.17 ±2.24c 0.139 ±0.006c 4712.0 ±87.0c
Methanol 24.77 ±1.24b 0.027 ±0.004a ND
Aqueous 9.63 ±2.60a 0.014 ±0.003a ND
Positive control
Rutin 649.93 ±13.34 1.789 ±0.214 42.7 ±2.3
Quercetin 1275.62 ±56.03 14.444 ±0.934 22.2 ±0.9
Results are presented as means ±SD (n=3).
IC50 values are presented for the DPPH radical scavenging activity.
Values within the same column with different letters (a–d) are significantly different at P<0.05 from the different extracts. ND, not detected.
FRAP and DPPH radical scavenging activity. Statistical Package
for the Social Sciences (SPSS) version 18.0 (Chicago, IL, USA) and
Microsoft Excel 2007 (Roselle, IL, USA) were used for the statistical
and graphical evaluations.
Total phenolic content
Phenolics are secondary metabolites that are ubiquitously present
in plants. Positive correlations between antioxidant activities
present in medicinal plants with their total phenolic content have
been reported. In our study, the phenolic values of P. crispum
extracts ranged from 9.63 ±2.60 to 42.31 ±0.50 mg GAE g1
(Table 1). The dichloromethane extract displayed the highest
phenolic content (P<0.05) among the extracts. The nature of
the extracting solvent is one of the most important factors in the
extraction of antioxidants, thus explaining the various phenolic
values from different extracts of P. crispum leaves and stems.
A study by Luthria22 investigated the influence of particle size on
phenolic compound extraction of P. crispum with ethanol:water,
50:50 (v/v), using a pressurized liquid extractor. They reported
phenolic values ranging from 18.3 to 22.9 mg GAE g1. In our
study, we showed higher phenolic contentin the dichloromethane
(42.31 ±0.50 mg GAE g1), ethyl acetate (32.17 ±2.24 mg GAE g1)
and methanolic (24.77 ±1.24 mg GAE g1)extractsofP. crispum
compared to that reported by Luthria.22 The dichloromethane
extract in our study also showed higher phenolic content com-
paredtotheP. crispum hydrodistilled ex tract (29.2 ±0.44 mg GAE
g1) reported by Hinneburg et al.23 A possible explanation for
the different phenolic values between these studies and that
reported in our study is that phenolic content in plants differ very
much between cultivars of the same species and are influenced
by genetic factors and environmental conditions.14 In addition,
parameters such as solvent polarity, extraction procedures and
conditions can influence the extraction of phenolic compounds
from P. crispum.24 Our study showed that the dichloromethane sol-
vent resulted in the highest extraction of phenolics from P. crispum
leaves and stems.
Ferric reducing antioxidant power
The FRAP values of P. crispum extracts are presented in Table 1.
Among the extracts tested, the dichloromethane extract of P.
crispum exhibited the highest FRAP value (0.360 ±0.009 mmol g1,
Table 2. Correlation analysis of total phenolic content with antioxi-
dant activities of Petroselinum crispum extracts
Pearson correlation (r)value
P. cri s p u m (leaf and stem) 0.875*0.910*
TPC, total phenolic content; FRAP, ferric reducing antioxidant power;
DPPH, 1,1-diphenyl-2-picryl hydrazyl radical scavenging activity.
*Correlation is significant at the 0.01 level.
Pearson correlation analysis was performed to assess the rela-
tionship between phenolic content and ferric reducing activities
of the leaf and stem extracts. A statistically significant positive
correlation was identified between FRAP and phenolic content
of P. cr i s p um (r=0.875, P<0.01; Table 2). This indicates that phe-
nolic compounds present in P. crispum contributed to their ferric
reducing activities. The reductive ability of the extracts suggests
their ability to donate electrons to reduce ferric tripyridyltriazine
(Fe3+-TPTZ) to the ferrous complex (Fe2+-TPTZ). This implies that
P. crispum extracts may provide antioxidative protection from free
radicals in actual biological systems by donating electrons to rad-
icals and blocking radical chain reactions from causing diseases
related to chronic oxidative stress.25
DPPH radical scavenging activity
The dichloromethane extract of P. crispum showed the lowest IC50
value (3310.0 ±80.5 μgmL
1,P<0.05) compared to the other P.
crispum extracts (Table 1). In a study by Zhang et al.,26 essential
oil from P. crispum showed antioxidant activities in the 𝛽-carotene
bleaching assay (EC50 =5.12 mg mL1) and DPPH scavenging
assay (EC50 =80.21 mg mL1). Hinneburg et al.23 reported that
the P. cr i s p um hydrodistilled extract showed an IC50 value of
12.0 ±0.10 mg mL1in the DPPH scavenging assay. In our study,
the P. crispum extracts showed better DPPH radical scavenging
activity (Table 1) compared to the two studies mentioned above.
A strong and significant positive correlation was seen between
DPPH scavenging activity and phenolic content of P. crispum
(r=0.910, P<0.01; Table 2). This shows that phenolic compounds
of P. crispum could be responsible for the observed DPPH radical
scavenging activity, since these compounds can readily donate
hydrogen atoms to the radical. From the antioxidant study, it © 2015 Society of Chemical Industry J Sci Food Agric (2015)
Antioxidant, DNA-protective and anticancer properties of Petroselinum crispum
Highest percentage inhibition (%)
Cell line
yl acetate
Figure 1. Anti-proliferative activities of Petroselinum crispum extracts on cancer cell lines, MCF-7, MDA-MB-231 and HT-29. Results are presented as
means ±SD (n=3). Values within the same cell line with different letters (a–c) are significantly different at P<0.05 from the different extracts. Extract
concentration tested: 0 –500 μgmL
is observed that the dichloromethane extract displayed highest
phenolic content and FRAP value while exhibiting best DPPH
radical scavenging activity among the extracts of P. crispum leaves
and stems.
Antiproliferative activity
Extracts of P. crispum (0– 500 μgmL
1) were tested for their effect
on the proliferation of MCF-7, MDA-MB-231 and HT-29 cells,
using the MTT assay. Generally, extracts of P. crispum leaves
and stems exhibited weak cytotoxic activity with percent inhi-
bitions below 50% (Fig. 1). Among the five extracts analyzed,
the dichloromethane extract exhibited the best antiprolifera-
tive activity. At the highest concentration tested (500μgmL
the dichloromethane extract showed a percentage inhibition
of 48.4% ±1.8%, 25.5% ±3.0% and 49.9% ±1.0% on MCF-7,
MDA-MB-231 and HT-29 cells, respectively (Fig. 1). The ethyl
acetate, methanol and aqueous extracts showed less than 20%
inhibition, even at 500 μgmL
1of extract. The different cytotoxic
effects of the various extracts in this study suggest the impor-
tance of using solvents of differing polarity in order to extract
compounds with various polarities that contribute to different
biological activities of the plant extract. Each extract of P. crispum
behaved differently against the cell lines. The distinct effects of
these extracts may be due to the phytodiversity or different mech-
anisms associated with the compounds present in the extracts
and the various susceptibility levels of cell lines to the plant
In a study by Yoshikawa et al.,29 the methanolic extract from
the aerial parts of P. crispum (1 and 10 μgmL
have potent estrogenic activity and increased MCF-7 cell pro-
liferation. In our study, the methanol extract did not increase
MCF-7 cell proliferation but exhibited a very weak antiprolifer-
ative effect on MCF-7 cells. Our study on P. crispum leaves and
stems showed that the dichloromethane extract displayed best
antioxidant and antiproliferative activities. Hence we selected the
dichloromethane extract for further analysis.
DNA protective activity
Within living cells, reactive oxygen species are constantly being
generated as normal by-products of mitochondrial respiration.
Uncontrolled levels of reactive oxygen species can cause severe
damage to macromolecules, especially DNA, leading to degener-
ative diseases such as cancer.30 H2O2is an oxidizing agent which
produces reactive hydroxyl radicals that can induce strand breaks
associated with DNA damage.20 The comet assay is a quick, sim-
ple and sensitive method for the evaluation of DNA damage,
mainly single-strand and double-strand breaks in individual cells.
The comet tail length is associated with DNA damage. Greater tail
length signifies greater DNA damage.19
Cells pre-treated with the dichloromethane extract of P. crispum
at concentrations of 100 –400 μgmL
1showed a significant
dose-dependent increase in DNA protection (P<0.05) compared
to the control of H2O2treatment alone (Table 3). At 400 μgmL
extract pretreatment, DNA damage was reduced by 50.9% ±6.6%
J Sci Food Agric (2015) © 2015 Society of Chemical Industry EL-H Tang et al.
Table 3. Protection from H2O2-induced DNA damage in 3 T3-L1
fibroblasts pre-treated with Petroselinum crispum extract
P. cri s p u m dichloromethane
extract (μgmL
1) DNA protection (%)
100 19.0 ±6.1*
200 23.1 ±6.9*
300 37.9 ±7.8*
400 50.9 ±6.6*
Results are presented as means ±SD (n=3).
*P<0.05 compared to control (without extract treatment), as tested
by Student’s t-test.
compared to the control, indicating 50.9% DNA protection. The
high phenolic content in the dichloromethane extract of P. crispum
as shown in Table 1 may be responsible for the observed DNA
protective effect. A study on spices (ginger, caraway, cumin,
cardamom, star anise and fennel) has shown a strong positive
correlation between DNA protection and phenols.20 Phenolics in
P. crispum can lower H2O2levels or hydroxyl radicals by increasing
the levels of H2O2-detoxifying enzymes in cells, thus preventing
DNA damage.31 Studies have shown that supplementation of
diets with fresh P. crispum leaves can increase antioxidant capacity
of rat plasma,11 protect against mitochondrial oxidative damage
10 and decrease oxidative stress in humans.12
Our study shows that the P. crispum extrac t protected 3 T3-L1
fibroblasts against H2O2-induced DNA damage, suggesting that
appropriate addition of the herb in the daily diet might reduce
the effects of free radical-induced carcinogenesis, hence affording
some protection against cancer.
Inhibition of H2O2-induced MCF-7 cell migration using
the scratch motility assay
Metastasis is the most characteristic aspect of malignant neoplasm
and is the leading cause of the ineffectiveness of chemothera-
peutic drugs and cancer deaths. The scratch motility assay tests
the ability of P. crispum extracts to inhibit migration of can-
cer cells in the denuded area, thus indicating defense against
(a) (b) (c)
(d) (e) (f)
20 µm
20 µm 20 µm 20 µm
20 µm 20 µm
ct (µg mL
ntage inhibi
27 ± 6*
41 ± 4*
30 ± 13
26 ± 8*
18 ± 7
Figure 2. Effect of Petroselinum crispum dichloromethane extract on the inhibition of H2O2-induced MCF-7 cell migration in a denuded area using the
scratch motility assay. (A)Photographs of cell migration (a) in untreated cells and (b) in cells treated with P. c r i sp u m dichloromethane extract at 200 μgmL
(c) 300 μgmL
1, (d) 400 μgmL
1, (e) 500 μgmL
1and (f ) 600 μgmL
1after 24 h. (B) Percentage inhibition presented as means ±SD (n=3). Asterisk
represents *P<0.05 compared to the control (without extract) as tested by Student’s t-test. © 2015 Society of Chemical Industry J Sci Food Agric (2015)
Antioxidant, DNA-protective and anticancer properties of Petroselinum crispum
0 100 200 400 500 600 700 800
Live cell number
% of control
P. crispum dichloromethane extract (µg mL
% of control Live cell number
Figure 3. Trypan blue exclusion assay of MCF-7 cells treated with Petroselinum crispum dichloromethane extract. The results are presented as the number
of viable cells counted per well and the percent of viable cells relative to untreated control. The viability of untreated control cells was taken as 100%.
Results are presented as means ±SD (n=3). *P<0.05 compared to the control (without extract) as tested by Student’s t-test.
metastasis.32 H2O2was included in this experiment to induce
the proliferation and migration of MCF-7 cells. The concentra-
tion of H2O2(1 μmol L1) used in this assay has been previously
tested in our laboratory and showed increased cell migration and
In this study, the scratch motility assay displayed the ability
of P. crispum to suppress H2O2-induced migration of MCF-7 cells
in a denuded area (Fig. 2). Treatment with P. crispum extract at
300 μgmL
1resulted in the highest inhibition of MCF-7 migration
(41% ±4%). At higher concentrations of P. crispum extract, the
inhibitory effect on cell migration decreased; at the highest con-
centration tested, 600 μgmL
1, the inhibition of migration was
the lowest (18% ±7%). The dichloromethane extract of P. crispum
inhibited the migration of MCF-7 cells, but not in a directly pro-
portional manner to the concentration of extract. It is interesting
to note that the inhibition of proliferation induced by the extract
was highest at the highest concentration of 500 μgmL
1. As cell
migration is necessary for metastasis of cancer cells, inhibition
of migration is an indication of protection against metastasis.
Petroselinum crispum prevented migration of MCF-7 cells, thus
showing potential in preventing metastasis. The flavonoids
present in P. crispum, apigenin and luteolin9have been reported
as chemopreventive agents of metastasis due to their ability to
prevent tumor cell motility and invasion.33 Phenolics present
in P. crispum might lower H2O2levels or hydroxyl radicals by
increasing the levels of H2O2-detoxifying enzymes in cells such as
glutathione peroxidase,31 thus preventing cancer cell proliferation
2O2. Antioxidants present in P. crispum
can maintain H2O2levels in cells within physiological levels and
may be associated with the prevention of cancer cell proliferation
and migration.
Trypan blue dye exclusion
Trypan blue is taken up by dead cells that have lost their mem-
brane permeability barrier or dye exclusion capacity, while the
intact plasma membrane of live cells excludes the dye.34 To
assess the antiproliferative effect of P. crispum on MCF-7 cells,
trypan blue exclusion counts were conducted on cells treated
with 0– 800 μgmL
1of dichloromethane extract for 48 h. A sig-
nificant dose-dependent decrease in live cell number (P<0.05)
was observed in cells treated with the dichloromethane extract
of P. crispum compared to the control (Fig. 3). This is shown
by the lower number of live cells counted as the concentra-
tion of dichloromethane extract treatment increased. Using
the trypan blue exclusion assay, the percent of viable cells rel-
ative to untreated control was 55.04%±0.75% at 500 μgmL
of dichloromethane extract treatment, whereas at the highest
concentration of extract treatment (800 μgmL
1) cell viability
decreased to 30.10% ±1.48%, indicating the antiproliferative
activity of P. crispum dichloromethane extract on MCF-7 cells.
This antiproliferation profile by trypan blue exclusion assay fur-
ther confirmed the inhibitory effect of P. crispum on MCF-7 cell
proliferation analyzed using the MTT assay (Fig. 1).
DNA fragmentation analysis
DNA fragmentation is a hallmark of apoptosis. In agarose gel elec-
trophoresis, apoptotic cells demonstrate a characteristic DNA ‘lad-
der’ pattern at 200 bp intervals, while necrotic cells are observed
as a ‘smear’ of randomly degraded DNA.35 However, internucleoso-
mal DNA fragmentation is not universal as it may not always occur
during apoptosis.36
In this study, apoptotic DNA fragmentation was analyzed using
agarose gel electrophoresis. DNA isolated from untreated con-
trol cells exhibited one clear band that pointed to the pres-
ence of living cells with intact DNA strand (Fig. 4). A typical
DNA ladder pattern was not evident in MCF-7 cells treated with
dichloromethane extract of P. crispum (500 or 800 μgmL
24 or 48 h. Instead, a smear pattern of DNA fragmentation was
observed in the extract-treated cells, with more intense smear-
ing seen in cells treated for 48 h, compared to 24 h. The results
could indicate that P. crispum dichloromethane extract kills MCF-7
cells by necrosis in a time-dependent manner, where random DNA
fragmentation occurs through the release of lysosomal DNases to
form a ‘smear’ on agarose gels.37 Conversely, studies have reported
that MCF-7 cells can undergo apoptosis without showing DNA
fragmentation due to lack of caspase-3, which is responsible for
this feature.38,39 Thus, using the results from DNA fragmentation
J Sci Food Agric (2015) © 2015 Society of Chemical Industry EL-H Tang et al.
1000 bp
10000 bp
250 bp
500 bp
750 bp
1500 bp
400 bp
100 bp
200 bp
1000 bp
2000 bp
600 bp
800 bp
Figure 4. Electrophoresis of DNA extracted from MCF-7 cells after treatment with Petroselinum crispum dichloromethane extract. M1: 1 kb DNA ladder;
lane 1: DNA from cells of control; lane 2: DNA treated with 500 μgmL
1extract for 24 h; lane 3: DNA treated with 800 μgmL
1extract for 24 h; lane 4: DNA
treated with 500 μgmL
1extract for 48 h; lane 5: DNA treated with 800 μgmL
1extract for 48 h; M2: 100 bp DNA ladder.
alone would not be definitive to accurately ascertain the mode of
cell death (whether by apoptosis or necrosis, or both), in which
P. crispum kills MCF-7 cells.Further work will be needed for more
in-depth investigation into the mechanism of cell death induced
by P. crispum.
The dichloromethane extract of P. crispum leaves and stems
showed antioxidant activities and also inhibition of proliferation
and cell migration in MCF-7 cells. The extract also protected
against DNA damage induced by H2O2.
Regular addition of P. crispum in the daily diet as food or supple-
ments can help strengthen the antioxidant systems of the body
and reduce the effects of free radical-induced carcinogenesis, can-
cer and subsequent metastasis caused by prolonged and excessive
oxidative stress.
The study was supported by the University of Malaya Research
University grants RG004/09AFR, PS250/2010B and RG341/11HTM.
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... The average yield of parsley ranges from 74 to 167 t·ha −1 [2]. Among several applications of parsley, its pharmacological properties such as antifungal, hepatoprotective, gastroprotective, anticancer, and antibacterial activities are the most important [3,4]. All parts of this plant are useful, including as food, cosmetic, and pharmaceutical ingredients [5]. ...
... Fejes et al. [7] studied the phytochemical profile of PL and reported the presence of flavonoids such as kaempferol and quercetin and glycosylated flavones such as luteolin and apigenin. In addition, PL are rich in ascorbic acid, terpenes, apiin, carotenoids, and tocopherol [4]. Wong and Kitts [8] studied the antioxidant effects of parley leaves in vitro and proposed that essential oil plays a crucial role in the scavenging effect. ...
Full-text available
Parsley leaves (PL) are a rich source of many bioactive compounds and show many health-promoting properties. The aim of this study is to analyze the effect of the addition of PL to wheat flour on the physical, antioxidant, and sensory properties of wheat bread. Wheat flour was partially substituted with 0, 1, 2, 3, 4, and 5% PL. Bread dough was prepared using the direct method. Bread loaves were cooled, and then their volume, texture, color, total phenolic content, and antioxidant activity were evaluated. In addition, a sensory evaluation of bread was performed. Incorporation of PL into wheat decreased the bread volume and increased the crumb moisture but had little influence on the crumb texture. The crumb of the enriched bread was darker and greener compared with the control sample. PL addition increased the redness of the crumb as well. The total color difference for the enriched bread ranged from 9.3 to 21.4. According to the sensory evaluation, the amount of wheat flour added to PL should not exceed 3%. Such a kind of bread showed about a twofold higher level of phenolic compounds and enhanced antioxidant activity compared with the control bread. This study showed that powdered PL can be a valuable nutritional supplement to wheat bread. Future research should focus on the possibilities of fortifying various types of food with this additive.
... Indeed, parsley importance is attributed to its high vitamins concentration (mainly vitamin C), antioxidants [11], and some mineral elements such as iron [12] as well as volatile oils that play an important role in the pharmaceutical and food industries [13]. Parsley has been used medicinally since ancient times for possible medicinal qualities including antioxidative [14], anti-carcinogenic [15], antimicrobial, laxative, antihyperlipidemic, anticoagulant and antihepatotoxic [16]. Because of the many antioxidants present in this plant, a diet including fresh parsley leaf can significantly increase antioxidant capacity, which plays a special role in people nutrition [10,17,18]. ...
Production of functional food with high levels of selenium and other antioxidants is very valuable for human protection against different forms of oxidant stress. Among leafy vegetables parsley demonstrate the highest levels of antioxidants. Biochemical analysis and fluorimetric determination of selenium revealed that foliar biofortification of 4 parsley (Petroselinum crispum) cultivars and Mitsuba (Cryptotaenia japonica) with sodium selenate (25 mg L-1) resulted in the highest biofortification level in curly parsley cultivar Krasotka (102.9) which showed the highest leaf surface area, antioxidant activity (65 mg GAE g-1 d.w.) and flavonoids content (25.9 mg quercetin equivalent g-1 d.w.), and the increase by 1.4 times in carotene content and 1,5 times in total chlorophyll content. ICP-MS method of mineral composition evaluation recorded extremely high levels of B and Si in Mustuba, which increased due to Se supplementation by 1.23 and 1.46 times respectively. In a two-year experiment with control and Se-fortified, leafy parsley, cultivar Moskvichka reached high values of seed yield and viability, and seed Se content (6170 µg kg-1 d.w.). The results of the present investigation demonstrate high prospects of parsley and Mitsuba selenium biofortification for production of functional food with elevated levels of microelement and high antioxidant activity.
... Moreover, El-Zaeddi et al. [11], who evaluated the phenolic compound composition and the antioxidant activity in three Apiaceae species (coriander, dill and parsley), suggested a variable association between phenolic compound content and antioxidant activity, depending on the species and the tested assay. In contrast to these reports, several other studies identified phenolic compounds as significant contributors to the overall antioxidant activity of parsley plant parts [14,54,58,65], highlighting the important effect of the implemented assay, the extraction protocol, the plant part used for the extraction and the growing conditions. The antimicrobial properties of the tested parsley root extracts are presented in Tables 8 and 9. ...
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In the present study, the chemical profile and bioactive properties of the roots of turnip-rooted parsley (Petroselinum crispum spp. tuberosum) germplasm were evaluated. For this purpose, plants from seventeen parsley cultivars were grown in 6 L pots, and the obtained roots were analyzed in terms of nutritional value, chemical composition (tocopherols, sugars and organic and fatty acids) and bioactive content (antioxidant activity, phenolic compound composition and antimicrobial properties). Our results showed great variability in terms of the chemical composition and bioactive properties of root parsley germplasm. A higher fresh root yield was recorded for the common "Root parsley" common variety (164 g/pot), followed by the varieties "Osborne" (109 g/pot), "Sonata" (104 g/pot), "Kaśka" (104 g/pot) and "Halblange Berlinska" (103 g/pot), whereas the lowest yield was recorded for the "Hanacka" variety (69 g/pot). A significant variation was also observed in the nutritional value parameters: the roots of the "Sonata" genotype showed the highest fat content; "Arat", "Osborne" and "Olomuńcka" had the highest ash content; the "Alba" cultivar contained significantly higher amounts of carbohydrates; and the "Vistula" cultivar showed the highest energetic value. The only detected isoforms of vitamin E were α-and δ-tocopherols; content varied depending on the cultivar, although α-tocopherol was the most abundant compound in most cultivars, especially in the "Arat" cultivar. Sucrose was the most abundant free sugar detected, especially in the "Sonata" cultivar (16.96 g/100 g dw), followed by apiose (2.93-5.55 g/100 g dw), glucose (1.3-3.47 g/100 g dw) and fructose (1.37-3.03 g/100 g dw). Moreover, malic acid was the most abundant organic acid in most of the tested cultivars. Twenty-one individual fatty acids were identified in all the studied cultivars, with linoleic (47.9-57.1%) and palmitic acid (20.66-20.5%) being the most abundant. Nineteen individual phenolic compounds were tentatively identified, including three phenolic acids, fourteen flavonoids and two hydrolyzable tannins, while apigenin-O-pentoside-O-hexoside was the most abundant. The antioxidant activity differed between the tested assays (TBARS and OxHLIA), and the most effective cultivars for the TBARS assay ("Root parsley (common variety)" and "Berlinski Halblange Springer") were those with the lowest antioxidant activity for the OxHLIA assay after 120 min. Finally, in most cases, the root extracts were more efficient or similarly effective compared to the positive controls against the tested bacteria and fungi. In conclusion, our results provide information regarding the chemical characterization and the bioactivities of the roots of turnip-rooted parsley germplasm that could be further exploited in sustainable and diversified agro-ecosystems through the introduction of this species as a novel/complementary crop in the traditional farming systems of the Mediterranean basin. Citation: Petropoulos, S.A.; Fernandes, Â.; Finimundy, T.C.; Polyzos, N.; Pinela, J.; Ivanov, M.; Soković, M.; Ferreira, I.C.F.R.; Barros, L. The Bioactivities and
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The herbal plant Petroselinum crispum (P. crispum) (Mill) is commonly available around the world. In this study, the leaves of the herbal plant P. crispum were collected from the central region of Al-Kharj, Saudi Arabia, to explore their in vitro pharmacological activity. Essential oil from the leaves of P. crispum was isolated using the hydrodistillation method. The composition of P. crispum essential oil (PCEO) was determined using Gas chromatography-mass spectrometry (GC-MS). A total of 67 components were identified, representing approximately 96.02% of the total volatile composition. Myristicin was identified as the principal constituent (41.45%). The in vitro biological activity was assessed to evaluate the antioxidant, antimicrobial, and anti-inflammatory potential of PCEO. PCEO showed the highest antimicrobial activity against Candida albicans and Staphylococcus aureus among all the evaluated microbial species. In vitro anti-inflammatory evaluation using albumin and trypsin assays showed the excellent anti-inflammatory potential of PCEO compared to the standard drugs. An in silico study of the primary PCEO compound was conducted using online tools such as PASS, Swiss ADME, and Molecular docking. In silico PASS prediction results supported our in vitro findings. Swiss ADME revealed the drug likeness and safety properties of the major metabolites present in PCEO. Molecular docking results were obtained by studying the interaction of Myristicin with an antifungal (PDB: 1IYL and 3LD6), antibacterial (PDB: 1AJ6 and 1JIJ), antioxidant (PDB: 3NM8 and 1HD2), and anti-inflammatory (3N8Y and 3LN1) receptors supported the in vitro results. Therefore, PCEO or Myristicin might be valuable for developing anti-inflammatory and antimicrobial drugs.
... In ancient times, the herb was used not only for culinary and medicinal purposes but was also subjected to a variety of superstitious practices by the Greeks and ancient Romans 3 . The roots of parsley have long been used as a diuretic, while the seeds have been used to treat gastrointestinal disorders, diarrhoea, halitosis, kidney stones, inflammation, and amenorrhea [4][5][6] . Parsley is commonly used as a food additive in Saudi Arabia and has attracted much attention as a basic food additive due to its health benefits 7 . ...
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In this chapter, the traditional use, the phytochemical composition, and the pharmacological activities of African medicinal plants displaying antibacterial effects were reported. We have pooled together the plants and phytochemicals active in pathogens of the family Enterobacteriaceae, as well as Pseudomonas aeruginosa, Gram-positive bacteria, and Mycobacteria. We also identified potent antibacterial medicinal plants of Africa having other pharmacological activities such as anti-inflammatory, anticancer, anti-diabetic, central nervous system, cardiovascular, anti-parasitic, hepatoprotective, immunomodulatory, nephroprotective, reproduction and digestive systems, antiviral, and wound healing activities. The documented plants can be further investigated globally by scientists to develop new herbal drugs to combat various types of bacterial infections.
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Culinary herbs and spices are known to be good sources of natural antioxidants. Although the antioxidant effects of individual culinary herbs and spices are widely reported, little is known about their effects when used in combination. The current study was therefore undertaken to compare the antioxidant effects of crude extracts and essential oils of some common culinary herbs and spices in various combinations. The antioxidant interactions of 1:1 combinations of the most active individual extracts and essential oils were investigated as well as the optimization of various ratios using the design of experiments (DoE) approach. The 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays were used to determine the antioxidant activity, and MODDE 9.1® software (Umetrics AB, Umea, Sweden) was used to determine the DoE. The results revealed synergism for the following combinations: Mentha piperita with Thymus vulgaris methanol extract (ΣFIC = 0.32 and ΣFIC = 0.15 using the DPPH and FRAP assays, respectively); Rosmarinus officinalis with Syzygium aromaticum methanol extract (ΣFIC = 0.47 using the FRAP assay); T. vulgaris with Zingiber officinalis methanol extracts (ΣFIC = 0.19 using the ABTS assay); and R. officinalis with Z. officinalis dichloromethane extract (ΣFIC = 0.22 using the ABTS assay). The DoE produced a statistically significant (R2 = 0.905 and Q2 = 0.710) model that was able to predict extract combinations with high antioxidant activities, as validated experimentally. The antioxidant activities of the crude extracts from a selection of culinary herbs and spices were improved when in combination, hence creating an innovative opportunity for the future development of supplements for optimum health.
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Cancer is the second cause of death in the world and is foreseen to be responsible for about 16 million deaths in 2040. Approximately, 60% of the drugs used to treat cancer are of natural origin. Besides the extensive use of some of these drugs in therapies, such as those derived from the genus Taxus, a significant number of plants have revealed themselves as useful against cancer in recent years. The field of ethnobotany focuses on documenting traditional knowledge associated with plants, constituting a starting point to uncover the potential of new plant-based drugs to treat or prevent, in this case, tumour diseases and side effects of chemotherapy and radiotherapy. From a series of extensive ethnobotanical prospections across the Catalan linguistic area (CLA), we have recorded uses for 41 taxa with antitumour effects. The two most quoted botanical families are Asteraceae and Ranunculaceae, and the most frequently reported species is Ranunculus parnassifolius, a high-mountain species, which is widely collected for this purpose. The reported species have been used to treat an important number of cancer types, focusing on preventive, palliative, and curative uses, as well as to deal with the side effects of conventional treatments. Comparing our results in CLA with previous data available in the most comprehensive databases of pharmacology and a review of cytotoxicity assays revealed that for the several species reported here, there was no previous evidence of traditional uses against cancer. Despite the need for further analyses to experimentally validate the information presented here, combining traditional uses and phylogenetically-informed strategies to phytochemical and pharmacological research would represent new avenues to establish more integrative approaches, hence improving the ability to select new candidate taxa in cancer research.
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ABSTRACT The invasiveness and low survivability on the part of patients associated with cancer continue to raise global concern. Different approaches have been used in the treatment and restoration of normalcy in cancer patients. However, most of the therapeutic strategies employed are challenged with high cost, low efficacy, high toxicity, and multiple side effects. In recent times, emergent studies have provided evidence that functional foods and their bioactive components serve roles as potential agents in the prevention and treatment of cancers. Moreover, global interest has focused on how this chemoprevention potential of functional foods can be explored as plant-based medicines for drug development. Although, the literature is replete with the mechanism of chemoprevention elicited by individual components of functional foods, there are limited reports on their overall anti-cancer mechanisms.
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Cell migration and invasion are processes that offer rich targets for intervention in key physiologic and pathologic phenomena such as wound healing and cancer metastasis. With the advent of high-throughput and high content imaging systems, there has been a movement towards the use of physiologically relevant cell-based assays earlier in the testing paradigm. This allows more effective identification of lead compounds and recognition of undesirable effects sooner in the drug discovery screening process. This article will review the effective use of several principle formats for studying cell motility: scratch assays, transmembrane assays, microfluidic devices and cell exclusion zone assays.
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Interleukin 1β-converting enzyme-like proteases (caspases) are crucial components of cell death pathways. Among the caspases identified, caspase-3 stands out because it is commonly activated by numerous death signals and cleaves a variety of important cellular proteins. Studies in caspase-3 knock-out mice have shown that this protease is essential for brain development. To investigate the requirement for caspase-3 in apoptosis, we took advantage of the MCF-7 breast carcinoma cell line, which we show here has lost caspase-3 owing to a 47-base pair deletion within exon 3 of the CASP-3 gene. This deletion results in the skipping of exon 3 during pre-mRNA splicing, thereby abrogating translation of theCASP-3 mRNA. Although MCF-7 cells were still sensitive to tumor necrosis factor (TNF)- or staurosporine-induced apoptosis, no DNA fragmentation was observed. In addition, MCF-7 cells undergoing cell death did not display some of the distinct morphological features typical of apoptotic cells such as shrinkage and blebbing. Introduction of the CASP-3 gene into MCF-7 cells resulted in DNA fragmentation and cellular blebbing following TNF treatment. These results indicate that although caspase-3 is not essential for TNF- or staurosporine-induced apoptosis, it is required for DNA fragmentation and some of the typical morphological changes of cells undergoing apoptosis.
Seven men and seven women participated in a randomized crossover trial to study the effect of intake of parsley (Petroselinum crispum), containing high levels of the flavone apigenin, on the urinary excretion of flavones and on biomarkers for oxidative stress. The subjects received a strictly controlled diet low in flavones and other naturally occurring antioxidants during the 2 weeks of intervention. This basic diet was supplemented with parsley providing 3.73-4.49 mg apigenin/MJ in one of the intervention weeks. Urinary excretion of apigenin was 1.59-409.09 mu g/MJ per 24 h during intervention with parsley and 0-112.27 mu g/MJ per 24h on the basic diet (P < 0.05). The fraction of apigenin intake excreted in the urine was 0.58 (sE 0.16) % during parsley intervention. Erythrocyte glutathione reductase (EC; GR) and superoxide dismutase (EC; SOD) activities increased during intervention with parsley (P < 0.005) as compared with the levels on the basic diet, whereas erythrocyte catalase (EC and glutathione peroxidase (EC activities did not change. No significant changes were observed in plasma protein 2-adipic semialdehyde residues, a biomarker of plasma protein oxidation. In this short-term investigation, an overall decreasing trend in the activity of antioxidant enzymes was observed during the 2-week study. The decreased activity of SOD was strongly correlated at the individual level with an increased oxidative damage to plasma proteins. However, the intervention with parsley seemed, partly, to overcome this decrease and resulted in increased levels of GR and SOD.