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Wound-Healing Activity of Some Species of Euphorbia L.

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Some species of Euphorbia have been used as medicinal plants to treat wounds, and skin diseases, around the world. The solvents n-hexane, ethyl acetate, and methanol were used successively to prepare extracts of the aerial parts of E. characias subsp. wulfenii, E. helioscopia, E. macroclada, E. seguieriana subsp. seguieriana, and E. virgata. Linear incision, circular excision wound models and the hydroxyproline assay method were used to assess the wound-healing activity. The inhibition of the increase in capillary permeability induced by acetic acid was used to assay the anti-inflammatory activity. The methanol extract of the aerial parts of E. characias subsp. wulfenii showed statistically significant wound-healing activity with 43.03% tensile strength for the linear incision wound model and a 65.24% reduction in the area of the wound by day 10 for the circular excision model. The tissue treated with this extract was found to contain 35.47 µg/mg of hydroxyproline. The methanol extract of E. characias subsp. wulfenii inhibited inflammation induced by acetic acid with a value of 34.74%. The results showed that the aerial parts of E. characias subsp. wulfenii possess wound-healing and anti-inflammatory activities on different models.
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ORIGINAL ARTICLE
The article was published by ACG Publications
http://www.acgpubs.org/journal/records-of-natural-products © March-April 2019 EISSN:1307-6167
DOI: http://doi.org/10.25135/rnp.81.18.03.255
Rec. Nat. Prod. 13:2 (2019) 104-113
Wound-Healing Activity of Some Species of Euphorbia L.
Serkan Özbilgin
1,*
, Esra Küpeli Akkol
2
, İpek Süntar
2
, Mehmet Tekin
3
and Gülçin Saltan İşcan
1
1
Department of Pharmacognosy, Faculty of Pharmacy, Ankara University, 06100, Ankara, Türkiye
2
Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, 06330, Ankara, Türkiye
3
Department of Pharmaceutical Botany, Faculty of Pharmacy, Trakya University, 22030, Edirne,
Türkiye
(Received March 15, 2018; Revised June 08, 2018; Accepted June 10, 2018)
Abstract: Some species of Euphorbia have been used as medicinal plants to treat wounds, and skin diseases,
around the world. The solvents n-hexane, ethyl acetate, and methanol were used successively to prepare extracts
of the aerial parts of E. characias subsp. wulfenii, E. helioscopia, E. macroclada, E. seguieriana subsp.
seguieriana, and E. virgata. Linear incision, circular excision wound models and the hydroxyproline assay method
were used to assess the wound-healing activity. The inhibition of the increase in capillary permeability induced by
acetic acid was used to assay the anti-inflammatory activity. The methanol extract of the aerial parts of E. characias
subsp. wulfenii showed statistically significant wound-healing activity with 43.03% tensile strength for the linear
incision wound model and a 65.24% reduction in the area of the wound by day 10 for the circular excision model.
The tissue treated with this extract was found to contain 35.47 µg/mg of hydroxyproline. The methanol extract of
E. characias subsp. wulfenii inhibited inflammation induced by acetic acid with a value of 34.74%. The results
showed that the aerial parts of E. characias subsp. wulfenii possess wound-healing and anti-inflammatory activities
on different models.
Keywords: Euphorbia; anti-inflammatory; excision; incision; wound-healing. © 2018 ACG Publications. All
rights reserved.
1. Introduction
The Euphorbia genus is the largest of the Euphorbiaceae plant family of about 2000 species,
ranging from annuals to trees. Euphorbia species contain latex and have very different flower states. An
important part of these are mostly originated from Africa or Madagascar. Ninety-one species of
Euphorbia grow in Turkey [1]. Some species of Euphorbia which are commonly known as “sütleğen”
in Turkish or “spurge” in English are used in traditional medicine to treat skin diseases, wounds, warts,
gonorrhea, migraines, and intestinal parasites in Turkey and other parts of the world [2]. The latex of
Euphorbia armena Prokh., and Euphorbia seguieriana subsp. seguieriana Necker, is used to treat
wounds and warts on the skin. The flowers of Euphorbia virgata Waldst. & Kit. are used on eczema [3-
5], Euphorbia fusiformis Buch.-Ham. ex. D.Don, Euphorbia helioscopia L., Euphorbia peplus L.
Euphorbia hirta L., and Euphorbia characias L. are used as antimicrobial and antifungal agents [6-10].
Ethnobotanical studies have shown that other species of this genus, such as Euphorbia macroclada L.
*
Corresponding author: E-Mail: ozbilgin@pharmacy.ankara.edu.tr; Phone:0090-312-2033103 Fax:0090-312-
2131081
Ozbilgin et.al., Rec. Nat. Prod. (2019) 13:2 104-113
105
and Euphorbia coniosperma Boiss., were used to enhence the healing of wound and also against
scorpion and snake bites [4,11].
Plants of the genus Euphorbia have been found to contain diterpenoids with the basic skeletons
of jatrophane, lathyrane, and myrsinane [12]; ingenane, daphnane and tigliane [13]; paraliane, pepluane,
and segetane. The sesquiterpenoids clovandiol, euphanginol, euphorbioside A, and euphorbioside B
have also been reported [14]. The flavonoids kaempferol, myricetin, rutin, quercetin, and their
derivatives and phenolic compounds have been found [15], along with the volatile compounds α-
terpineol, β-caryophyllene, α-humulene, linalool, terpinene, and germacrene-D [16]. Tannins
(euphorbins), triterpenoids (lupeol, lupeol acetate, betulin, and β-amyrin) and phytosterols such as β-
sitosterol have also been reported [17].
Euphorbia species have an increasingly prevalent due to chemical compounds which have
different skeletal structure and their therapeutic importance. The results showed that Euphorbia species
have cytotoxic, antitumor, antibacterial, anti-inflammatory and anti-HIV activities [18]. Euphorbia
species also show analgesic, antidiorrheal, antifeedant, antimicrobial, antiproliferative, antipyretic, and
molluscicidal activities and the ability to modulate multidrug resistance [13-14].
The existing ethnobotanical studies covering traditional uses of Euphorbia species do not
provide scientific proof of their effectiveness. The aim of the present study is to investigate the in vivo
wound-healing and anti-inflammatory potentials of five Euphorbia species, some of which have been
used in traditional medicine. The species E. characias subsp. wulfenii (Hoppe ex W. Koch) A. R. Smith,
E. helioscopia, E. macroclada, E. seguieriana subsp. seguieriana, and E. virgata were chosen. Extracts
of the aerial parts of the selected species were prepared by using n-hexane, ethyl acetate, and methanol.
Linear incision, circular excision wound models and the hydroxyproline assay method were used to
assess the wound-healing activity. The inhibition of the increase in capillary permeability induced by
acetic acid method was used to assay the anti-inflammatory activity.
2. Materials and Methods
2.1. Plant Material
Plants of various species of Euphorbia were collected from different regions of Anatolia, Turkey
(Table 1). Taxonomic identification of the plants was confirmed by Prof. Dr. Hayri Duman at
Department of Biology, Faculty of Sciences, Gazi University, and Assoc. Prof. Mehmet Tekin at
Department of Pharmaceutical Botany, Faculty of Pharmacy, Trakya University. Voucher specimens
have been deposited in the herbarium at the Faculty of Pharmacy of Ankara University (AEF) and in the
herbarium at the Faculty of Science of Cumhuriyet University (CUFH). The localities where the plant
materials were collected and the dates when they were collected are shown along with the assigned
herbarium numbers in Table 1.
Table 1. Locality of the plants sample
Plant species Locality Altitude Date Herbarium No
E. macroclada
Çayyolu village, Yenimahalle,
Ankara 900 m 10.07.2012 AEF 26268
E. helioscopia Çomaklı village, Korkuteli, Antalya 1000 m 14.05.2012 AEF 26269
E. characias subsp.
wulfenii Yazır village, Korkuteli, Antalya 950 m 15.05.2012 AEF 26270
E. virgata Tödürge Lake, Zara, Sivas 1250 m 16.09.2012 M 1326
E. seguieriana subsp.
seguieriana İğneada, Kırklareli 250 m 21.08.2012 M 1321
2.1.1. Extraction
Air-dried and powdered aerial parts of each plant (80 g) were extracted with n-hexane (900mL
x 4) at room temperature during 8 h/day for 4 days with continuous stirring. The residues were dried
and extracted with ethyl acetate (900 mL x 4) at room temperature during 8 h/day for 4 days with
continuous stirring. These residues were dried and extracted with methanol (900 mL x 4) at room
Wound-healing activity of Euphorbia
106
temperature during 8 h/day for 4 days with continuous stirring. The extracts were filtered and the solvent
was removed to dryness at 50 °C under reduced pressure to yield the crude extracts.
2.2. Biological Activity Tests
2.2.1. Animals
Male Sprague Dawley rats (160–180 g) and Swiss albino mice (20–25 g) were purchased from
Laboratory of Experimental Animals, Kobay, Ankara, Turkey. The animals were acclimated for 3 days
in room conditions and fed a standard pellet diet and water ad libitum. Six animals from each group
were used to evaluate the activity tests. Throughout the experiments, the animals were processed
according to the suggested European ethical guidelines for the care of laboratory animals. The study
was performed according to the international rules covering animal experiments and biodiversity rights
(Gazi University Ethical Council Project Number: G.U.ET-12.049).
2.2.2. Preparation of the Test Samples
The test materials in an ointment base were applied topically to the wounded area of the test
animal to evaluate the wound-healing activity using the incision and excision wound models. The
extracts were mixed thoroughly in a mortar with a mixture of glycol stearate: propylene glycol and liquid
paraffin (3:6:1) to form an ointment with a concentration of 1%. The animals allocated to the control
group were treated topically with the blank vehicle, while the animals in the negative control group were
not treated with anything. Madecassol® (Bayer) (0.5 g) was used topically as a reference drug.
Madecassol® contains a 1% extract of Centella asiatica L. [19].
Test samples suspended in a mixture of distilled water and 0.5% sodium carboxy methyl
cellulose (CMC) were given to the test animals orally to evaluate anti-inflammatory activity. Animals
in the control group received only the vehicle [20]. Indomethacin (10 mg/kg) in 0.5% CMC was used
as a reference drug [21].
2.2.3. Wound Healing Activity
2.2.3.1. Linear Incision Wound Model
Ketasol® (Richterpharma) with a dose of 0.15 cc were used to anesthetize the animals. The
dorsal sides of the rats were shaved and disinfected with 70% ethanol. Two linear-paravertebral
incisions, each 5 cm long and 1.5 cm from the dorsal midline were made through the shaved skin using
a sterile blade. Three surgical sutures were each placed 1 cm apart.
The test ointments, the reference drug, or the ointment base (vehicle) was applied topically on
the wounds once a day for 9 days. The sutures were removed on day ten and the tensile strengths of the
injured and treated skin were measured with a tensiometer (Zwick/Roell Z 0.5, Germany) [22-23].
2.2.3.2. Circular Excision Wound Model
The circular excision wound model was used to evaluate the wound contraction and wound
closure time. Each animal group was anesthetized using 0.01 cc of Ketasol® (Richterpharma). After the
dorsal parts of the animals had been shaved, a circular wound was created on application area of each
mouse by excising the skin with a 5 mm biopsy punch (Nopa instruments, Germany) and nothing was
applied on the wounds [24]. The test samples, the reference drug (Madecassol®, Bayer), and the vehicle
ointments were applied topically once a day until the wound was completely healed. The wound areas
were photographed every day using a camera (Fuji, S20 Pro, Japan). The wound area was evaluated
using the program AutoCAD. The wound contraction was calculated as a percentage of the reduction in
the area covered by the wound [23,25].
2.2.3.3. Estimation of Hydroxyproline
Isolated tissues were dried at 60-70 °C in a hot-air oven until a consistent weight was achieved.
Each sample was hydrolyzed with 6 N HCl for 3 h at 130 °C. The solution was adjusted to pH 7 and
Ozbilgin et.al., Rec. Nat. Prod. (2019) 13:2 104-113
107
subjected to chloramin T oxidation. Absorbance of the colored adduct-product formed with Ehrlich
reagent at 60 °C was read at 557 nm using a Beckmann Dual Spectrometer (Beckman, Fullerton, CA,
USA). Standard hydroxyproline was also run and values reported as µg/mg dry weight of tissue [26,
27].
2.2.4. Anti-inflammatory Activity
The effects of the test samples on the increased vascular permeability induced by acetic acid in
mice was determined according to the Whittle method [28]. Test samples were orally administered with
0.2 mL/20 g body weight doses. 0.1 mL of 4% Evans blue in saline solution was injected to the tails of
each animal thirty minutes later. After 10 min 0.4 mL of 0.5% (v/v) acetic acid was injected
intraperitoneally. The mice were sacrificed 20 min later by dislocation of the neck, and the viscera was
washed with distilled water, then poured into 10 mL volumetric flasks from glass wool. Each flask was
made up to 10 mL with distilled water, 0.1 mL of 0.1 N NaOH solution was added, and the absorption
of the final solution was measured at 590 nm using a Beckmann Dual Spectrometer (Beckman,
Fullerton, CA, USA). The control group was given a mixture of oral water and 0.5% CMC.
2.3. Statistical Analysis of the Data
One-way analysis of variance (ANOVA) and Students-Newman-Keuls post hoc tests were used
to analyze the data. Values of p ˂ 0.05 were considered statistically significant.
3. Results and Discussion
Wound-healing and anti-inflammatory activity results of n-hexane (Hex), ethyl acetate (EtOAc), and
methanol (MeOH) extracts prepared from the aerial parts of E. characias subsp. wulfenii, E.
helioscopia, E. macroclada, E. seguieriana subsp. seguieriana, and E. virgata, are shown in the tables.
Table 2. Effects of the test materials on the linear incision wound model and the hydroxyproline
content of each extract
Material Extract
Tensile strength of wound
± SEM
(tensile strength %)
Hydroxyproline (µg/mg)
± SEM
Vehicle 13.34±1.79 (2.30) 12.47±2.15
Negative control 13.04±1.87 (-) 9.66±2.51
E. virgata
Hex
14.07±1.38 (5.47)
13.01
±2.70
EtOAc
14.61±1.42 (9.52)
12.36
±2.02
MeOH 16.42±1.56 (23.09) 17.85±2.17
E. macroclada Hex 13.08±1.41 (-) 14.78±2.12
EtOAc
13.87±1.29 (3.97)
11.06
±2.33
MeOH
17.35±1.17
(30.06**)
28.37
±
1.59**
E. seguieriana
subsp. seguieriana Hex 14.02±1.46 (5.09) 10.28±2.19
EtOAc
14.58±1.15 (9.30)
10.92
±2.26
MeOH
16.50±1.39 (23.69)
20.74
±1.99
E. characias
subsp. wulfenii Hex 14.17±1.52 (6.22) 9.67±2.13
EtOAc
16.11±1.79 (20.76)
20.62
±2.01
MeOH
19.08±1.47
(43.03**)
35.47
±
1.38***
E. helioscopia Hex 14.27±1.58 (6.97) 11.73±2.07
EtOAc 15.12±1.70 (13.34) 15.08±2.11
MeOH
18.35±1.84
(37.56**)
30.64
±
1.44**
Madecassol
®
21.41±1.17
(60.49***)
47.61
±
1.13***
SEM: Standard Error of the Mean; Hex- hexane extracts; EtOAc- ethyl acetate extracts; MeOH- methanol
extracts;
* : p < 0.05;
** : p < 0.01;
*** : p < 0.001
Wound-healing activity of Euphorbia
108
The methanol extract of E. characias subsp. wulfenii showed more wound-healing activity than
the other extracts, with a 43.03% tensile strength value for the linear incision wound model (Table 2)
and a 65.24% reduction of the wound area at day 10 for the circular excision wound model (Table 3).
The tissue treated with the methanol extract of E. characias subsp. wulfenii was found to contain 35.47
µg/mg of hydroxyproline (Table 2).
Table 3. Effects of the test materials on the circular excision wound model
Material Extract
Day
Wound area (mm
2
) ± SEM (Contraction %)
0 2 4 6 8 10
Vehicle 19.89±2.31 17.32±2.37
-
15.29±2.42
-
12.71±1.96
(1.93)
6.49±1.49
(8.46)
3.05±1.25
(10.03)
Negative Control 19.21±2.19 17.15±2.24 15.21±2.30 12.96±1.77 7.09±1.57 3.39±1.82
E. virgata
Hex 19.33±2.02 17.48±1.84
-
5.02±1.47
(1.77)
13.02±1.38
-
6.19±1.03
(4.62)
2.95±0.41
(3.28)
EtOAc 19.53±2.11 16.91±1.96
(2.37)
14.78±1.29
(3.34)
12.82±1.26
-
5.82±1.07
(10.32)
2.98±0.66
(2.29)
MeOH 19.38±2.25 15.88±1.82
(8.31)
14.51±1.25
(5.10)
11.91±1.17
(6.29)
5.74±0.98
(11.56)
2.23±0.71
(26.89)
E. macroclada
Hex 19.62±2.28 16.34±1.61
(5.66)
14.92±2.10
(2.42)
12.85±1.40
-
6.04±1.51
(6.93)
2.97±0.98
(2.62)
EtOAc 19.45±2.17 15.01±1.45
(13.34)
13.95±1.98
(8.76)
11.72±1.36
(7.79)
5.72±0.60
(11.86)
2.47±0.49
(19.02)
MeOH 19.47±2.26 14.37±1.58
(17.03)
12.99±1.79
(15.04)
10.87±1.44
(14.48)
4.44±0.35
(31.59)*
1.71±0.77
(43.93)**
E. seguieriana
subsp.
seguieriana
Hex 19.49±2.14 17.42±1.79
-
15.04±1.59
(1.64)
11.24±1.42
(11.57)
5.84±1.12
(10.01)
2.86±0.81
(6.23)
EtOAc 20.02±2.03 16.49±1.81
(4.79)
16.42±1.10
-
11.32±1.45
(10.94)
5.76±1.13
(11.25)
2.59±0.89
(15.08)
MeOH 19.99±2.22 16.32±1.85
(5.77)
15.44±1.47
-
11.43±1.55
(10.07)
4.89±1.08
(24.65)
2.46±0.92
(19.34)
E. characias
subsp. wulfenii
Hex 19.32±2.01 16.85±1.43
(2.71)
14.18±1.12
(7.26)
10.17±1.39
(19.98)
5.74±1.22
(11.56)
2.72±0.49
(10.82)
EtOAc 19.27±2.10 16.39±1.59
(5.37)
13.41±1.19
(12.29)
9.99±1.43
(21.40)
4.52±0.89
(30.35)
1.95±0.58
(36.07)*
MeOH 21.03±2.11 15.11±1.34
(12.76)
12.88±1.23
(15.76)
9.90±1.57
(22.11)
4.36±0.92
(32.82)*
1.06±0.39
(65.24)**
E. helioscopia
Hex 19.57±2.04 17.25±1.28
(0.40)
14.25±1.31
(6.80)
11.52±1.37
(9.36)
5.87±0.97
(9.55)
3.22±0.71
-
EtOAc 19.31±2.07 16.81±1.19
(2.94)
14.06±1.43
(17.20)
10.84±1.30
(16.28)
4.92±0.81
(24.19)
2.35±0.65
(22.95)
MeOH 20.10±2.31 15.71±1.23
(9.29)
14.01±1.25
(8.37)
10.79±1.53
(15.11)
4.02±0.74
(38.06)*
1.72±0.32
(43.61)**
Madecassol® 19.81±2.05 14.28±1.30
(17.55)
12.20±1.37
(20.21)
6.82±1.24
(46.34)**
1.85±0.62
(71.49)**
0.00±0.00
(100.00)***
SEM: Standard Error of the Mean; Hex- hexane extracts; EtOAc- ethyl acetate extracts; MeOH- methanol
extracts
* : p < 0.05
** : p < 0.01
*** : p < 0.001
Anti-inflammatory activity test results showed that, the methanol extracts of E. characias subsp.
wulfenii, and E. macroclada inhibited inflammation by 34.74% and 38.81%, respectively (Table 4). The
Ozbilgin et.al., Rec. Nat. Prod. (2019) 13:2 104-113
109
wound-healing and anti-inflammatory activity tests were consistent. The methanol extract of E.
characias subsp. wulfenii demostrated remarkable bioactivity in both the wound-healing and acute
inflammation models.
Table 4. Inhibitory effect of the test materials on the acetic acid-induced increase in capillary
permeability
Material Extract Dose (mg/kg) Evans Blue Concentration
(g/mL) ± SEM Inhibition (%)
Control 12.29±1.25
E. virgata
Hex
100
12.41±1.02
-
EtOAc 100 11.85±1.27 3.58
MeOH
100
10.32±1.21
16.03
E. macroclada
Hex 100 12.75±1.33 -
EtOAc 100 11.47±1.39 6.67
MeOH 100 7.52±0.93 38.81***
E. seguieriana
subsp. seguieriana
Hex
100
13.04±1.76
-
EtOAc
100
11.31±1.49
7.97
MeOH 100 10.75±1.05 12.53
E. characias subsp.
wulfenii
Hex 100 11.95±1.34 2.77
EtOAc 100 9.24±0.85 24.82
MeOH 100 8.02±0.79 34.74**
E. helioscopia
Hex 100 10.99±0.91 10.58
EtOAc
100
9.91±0.74
19.37
MeOH 100 9.45±1.02 23.11
Indomethacin 10.0 6.81±0.37 44.59***
SEM: Standard Error of the Mean; Hex- hexane extracts; EtOAc- ethyl acetate extracts; MeOH- methanol
extracts
* : p < 0.05
** : p < 0.01
*** : p < 0.001
Collagen is the major protein of the extracellular matrix. It makes the hydroxyproline and its
peptides free up [29]. The rate of collagen synthesis and the maturation process where covalent binding
of collagen fibrils are deterministic for a tensile strength of a wound [30]. In the present study, the tensile
strength of the new tissue is better than the treated groups and the signs of infection were shown at least
on level. The tensile strength is the resistance to breakage shown against stretching. This shows how
much the repaired tissue is resistanted against breaking. For this purpose, the newly repaired tissue was
removed and the tensile strength was measured [22]. Large amounts of hdyroxyproline in tissues
indicate the presence of collagen and accelerated healing [31]. The amount of hyroxyproline indicated
by the wound tensile strength parallels the results of incision wound model in this study.
The first phase of healing is inflammation. However, an elongated inflammatory response
delays healing. For this purpose, anti-inflammatory activity of the extracts was evaluated by using the
Whittle method. This model of inflammation determines the effectiveness of a test material against the
increased capillary permeability induced by acetic acid [32]. In the present study, the methanol extracts
of E. characias subsp. wulfenii, and E. macroclada significantly inhibited this inflammation more than
other groups.
Anti-inflammatory activity has been reported for Euphorbia australis Boiss., E. drummondii
Boiss., E. heyneana Spreng, E. hirta, E. kansui T.N. Liou ex. T.P. Wang, and E. royleana Boiss [33-
37]. E. fusiformis, E. helioscopia, and E. segetalis L. have shown antimicrobial activity [8,38,39].
Antioxidant activity has been attributed to E. helioscopia, E. hirta, E. macroclada, and E. rigida M.
Bieb. [40-42], and wound-healing activity to E. caducifolia Haines., E. hirta, and E. neriifolia L. [43,44].
The results obtained are similar to the wound-healing potential of the latex of E. caducifolia which
accelerated closure of the wound with greater contents of fibroblasts and collagen in the treated animals
[45]. The same type of wound-healing effect has also been observed for E. neriifolia [46]. Ahmed et al.
[47] have suggested that the topical administration of ethyl acetate and methanol extracts of E.
Wound-healing activity of Euphorbia
110
consobrina N.E.Br., ethyl acetate extract of E. inarticulata Schweinf., and methanol extracts of E.
balsamifera Aiton, and E. schimperi C. Presl. have significant therapeutic effects on the various phases
involved in the process of wound contraction and healing. Badgujar et al. [48] have reported that the
latex of E. nivulia Buch.-Ham. which included alkaloids, cynogenic glycosides, phenolics, saponins,
and tannins significantly reduced the bleeding and whole-blood clotting times. Results of the
ethnobotanical studies show that Euphorbia species have been used such as analgesic, anti-
inflammatory, antifungal, antiviral, cytotoxic, diuretic, laxative, and wound healer agents in traditional
medicine. However, there is not enough satisfactory studies which provide scientific evidence to prove
the effectiveness of these uses.
The aerial parts and latex of Euphorbia species are reputed to have various biological effects
that make them useful in traditional medicines. Giordiani et al. [7] have reported that plants of the
Euphorbia genus showed significant antimicrobial activity against Gram-positive and Gram-negative
bacteria. Such antimicrobial activity is particularly important because of it prevents infection of the
wounded area during the healing process. Free radicals are known to induce cell damage by lipid
peroxidation. Antioxidant activity therefore also contributes to the healing process. The antioxidant
activities of the Euphorbia species, E. heyneana, E. hirta, E. macroclada, and E. rigida have been
investigated [34, 40, 49]. Phytochemicals such as alkaloids, triterpenoids, tannins, phenolic compounds
and flavonoids are also known to support healing process, especially because of antimicrobial and
astringent properties. Flavonoids and their derivatives have been known to prevent or slow down the
progression of cell necrosis by increasing vascular formation of the tissues. Flavonoids are also thought
to be effective compounds responsible for wound contraction and epithelialisation.[50-54]. The genus
Euphorbia has been reported to be a rich source of sesquiterpenoids, glycerols, cerebrosides,
phloracetophenones, steroids, phenolic compounds and flavonoids [14]. In our study, significant wound-
healing and anti-inflammatory effects of the extracts are attributed to flavonoids and quercetin
glycosides. Flavonoid and flavonoid derivative contents of our selected species are consistent with
studies in literature. Previously quercetin-3-O-glucoside, kaempferol, kaempferol-3-O-glucoside,
kaempferol-3-rutinoside and rutin were isolated from E. virgata [13]. Ertaş et al. [55] have reported that
E. macroclada contained rutin, hyperoside, quercetin, apigenin, kaempferol, myricetin, naringenin and
hesperetin. In biological activity study report published by Pisano et al. [56], researchers have been
showed phytochemical profile of E. characias by using LC-MS/MS. Results of the study have been
showed that E. characias contain flavonoids namely quercetin-3-O-glucoside, quercetin-3-O-
rhamnoside, quercetin-3-O-arabinoside and quercetin-3-O-xyloside, which have antioxidant and
antimicrobial activities.
In the present study, results have shown that the methanol extract proposed from the aerial parts
of E. characias subsp. wulfenii possesses the best wound-healing activity of any of the five species of
Euphorbia we tested using three different solvents for extraction and it also outperformed the control
groups. This is attributed to the combined effect of the constituents present in extracts, especially the
flavonoids (see supporting information). The present study provides evidence to support the traditional
use of the aerial parts of E. characias subsp. wulfenii for wound healing.
Acknowledgements
This work was financially supported by the Ankara University Scientific Research Projects
Coordination Unit [Project Number: 13L3336004].
Supporting Information
Supporting Information accompanies this paper on http://www.acgpubs.org/journal/records-
of-natural-products
ORCID
Serkan Özbilgin: 0000-0002-3945-6756
Esra Küpeli Akkol: 0000-0002-5829-7869
İpek Süntar: 0000-0003-4201-1325
Ozbilgin et.al., Rec. Nat. Prod. (2019) 13:2 104-113
111
Mehmet Tekin: 0000-0002-6504-2223
Gülçin Saltan İşcan: 0000-0001-6633-0713
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