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Thrombin Inhibition: Preliminary Assessment of the Anticoagulant Potential of Turnera subulata (Passifloraceae)

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Cardiovascular and thromboembolic disturbances are the main causes of disease-related deaths worldwide. Regardless of the etiological factors involved in thrombus formation, coagulation is mainly activated by thrombin, one of the most important blood clotting molecules. Thus, this study evaluated the Turnera subulata leaf crude extract, its ethyl acetate fraction effect on the coagulation cascade, and its possible side effects. Their phytocomposition indicated polyphenols, mainly flavonol-3-O-glycosylate and a flavone glycoside, without in vitro and in vivo toxicity. Regarding their potential anticoag-ulants, results displayed partial thromboplastin and prothrombin time activation, and Xa and IIa, and thrombin inhibition by heparin II cofactor, indicating significant anticoagulant activity, suggesting direct and indirect thrombin inhibition as the main mechanism of action. Therefore, T. subulata leaf active compounds exhibit therapeutic potential required to develop phy-totherapeutic formulations to assist conventional anticoagulants in clinical treatments.
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Thrombin Inhibition:
Preliminary Assessment of the Anticoagulant Potential
of Turnera subulata (Passifloraceae)
Jefferson Roma
´ryo Duarte da Luz,
1,2
Thayse Evellyn Silva do Nascimento,
2
Leandro Vinicius Fernandes de Morais,
2
Ana Katarina Menezes da Cruz,
3
Adriana Augusto de Rezende,
1,2
Jose
´Branda
˜o Neto,
4
Marcela Abbott Galva
˜o Ururahy,
2
Andre
´Ducati Luchessi,
1,2
Jorge A. Lo
´pez,
5
Hugo Alexandre Oliveira Rocha,
3
and Maria das Grac¸as Almeida
1,2
1
Postgraduation Program in Health Sciences, Health Sciences Center,
Federal University of Rio Grande do Norte, Natal, Brazil.
2
Multidisciplinary Research Laboratory, DACT, Health Sciences Center,
Federal University of Rio Grande do Norte, Natal, Brazil.
3
Department of Biochemistry, Biosciences Center, Federal University of Rio Grande do Norte, Natal, Brazil.
4
Department of Clinical Medicine, Health Sciences Center, Federal University of Rio Grande do Norte, Natal, Brazil.
5
Tiradentes University/Institute of Technology and Research, Aracaju, Brazil.
ABSTRACT Cardiovascular and thromboembolic disturbances are the main causes of disease-related deaths worldwide.
Regardless of the etiological factors involved in thrombus formation, coagulation is mainly activated by thrombin, one of the
most important blood clotting molecules. Thus, this study evaluated the Turnera subulata leaf crude extract, its ethyl acetate
fraction effect on the coagulation cascade, and its possible side effects. Their phytocomposition indicated polyphenols, mainly
flavonol-3-O-glycosylate and a flavone glycoside, without in vitro and in vivo toxicity. Regarding their potential anticoag-
ulants, results displayed partial thromboplastin and prothrombin time activation, and Xa and IIa, and thrombin inhibition by
heparin II cofactor, indicating significant anticoagulant activity, suggesting direct and indirect thrombin inhibition as the main
mechanism of action. Therefore, T. subulata leaf active compounds exhibit therapeutic potential required to develop phy-
totherapeutic formulations to assist conventional anticoagulants in clinical treatments.
KEYWORDS: clinical therapy coagulation cascade plant extract polyphenols
INTRODUCTION
Nowadays, cardiovascular and thromboembolic
disorders are leading causes of death worldwide.
1,2
Arterial thrombosis is the most common cause of acute
myocardial infarction, stroke, and ischemia, while deep
venous thrombosis complications include pulmonary
embolism and post-thrombotic syndrome.
3
Thereby, the
coagulation system and its interaction with platelet ag-
gregation are responsible for arterial and venous thrombus
formation.
4
Regular indications for anticoagulant uses include pro-
phylaxis and venous thromboembolism treatments, cardi-
oembolic prevention in patients with cardiac arrhythmia or
mechanical valve prostheses, as well as secondary pre-
vention in patients with acute coronary syndromes or un-
dergoing percutaneous coronary intervention.
5
At this
point, nonfractionated and low molecular heparins are
used as anticoagulants, even causing side effects, such as
thrombocytopenia and a high risk of systemic bleeding.
6,7
This stimulates the search for new substances to aid in
prolonged anticoagulant therapy.
8
Thus, plant extracts exhibit a proven ability to inhibit the
blood coagulation cascade, especially regarding intrinsic and
extrinsic pathway factors.
9–12
In this context, Turnera sub-
ulata (family Passifloraceae) is widely distributed in tropical
and subtropical regions and used in folk medicine
12
due to its
pharmacological properties, such as anti-inflammatory,
13,14
hypoglycemic,
15
antifungal,
16
and antioxidant properties.
17
Phytochemical studies with species of this genus revealed the
presence of flavonols, alkaloids, tannins, cyanogenic gly-
cosides, fatty acids, triterpenoids, and various phenolic
compounds related to bioactivities.
18
Several studies describe the use of the Turneraceae genus
in inflammatory processes.
19
Thus, considering that thrombin
is closely related to coagulation and inflammatory systems,
Manuscript received 18 September 2018. Revision accepted 28 January 2019.
Address correspondence to: Maria das Grac¸as Almeida, PhD, Laborato
´rio Multi-
disciplinar em Pesquisa, Faculdade de Farma
´cia, Universidade Federal do Rio Grande
do Norte, R. Gen. Gustavo Cordeiro de Farias, s/n–Petro
´polis, Natal 59012-570, Brasil,
E-mail: mgalmeida84@gmail.com
JOURNAL OF MEDICINAL FOOD
J Med Food 00 (0) 2019, 1–9
#Mary Ann Liebert, Inc., and Korean Society of Food Science and Nutrition
DOI: 10.1089/jmf.2018.0141
1
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this genus represents a potential reservoir for discovering
compounds for clinical treatment associated with conven-
tional anticoagulants to minimize their side effects. There-
fore, this study evaluated the anticoagulant potential and toxic
and hemorrhagic effects of the T. subulata leaf crude extract
and its fraction of ethyl acetate.
MATERIALS AND METHODS
Plant material and leaf extract preparation
T. subulata leaves, collected in Natal, Rio Grande do
Norte, Brazil, were taxonomically identified by Dr. Jomar
Gomes Jardim, depositing a voucher specimen (Herbarium
No: 0674/08) at the Herbarium of the Department of Botany
and Zoology, Federal University of Rio Grande do Norte,
Natal, RN, Brazil. Leaves were air-dried at 40C for 48 h
and powdered, before preparing its T. subulata crude extract
(CETS) by ethanol:water (50:50, v/v) maceration for 4 days,
filtered, and then lyophilized.
To characterize CETS-active compounds, an extract
portion, resuspended in methanol, was subjected to liquid–
liquid partition using increasing polarity solvents: n-hexane
(3 ·300 mL) and ethyl acetate (3 ·300 mL), obtaining two
fractions hexanic fraction of T. subulate (HFTS) and
acetate fraction of T. subulate (AFTS). Phenolic compound
contents were determined in all fractions by the Folin–
Ciocalteu method described
20
(data not shown). No poly-
phenols were detected in HFTS.
High performance liquid chromatography with diode
array chromatographic profile
Chromatographic analyses in triplicates were performed on
a Phenomenex C18 chromatography column (4.6 ·100 mm,
particle size 2.6 lm; Torrance, CA, USA) coupled to the
HPLC system (VARIAN ProStar, Walnut Creek, CA, USA)
equipped with a ProStar 240 quaternary pump, autosampler
(ProStar 410), and a detector (mod. 355 PDA UV/V). CETS
and AFTS (5 mg/mL) were dissolved in methanol.
17
Nine
microliters were injected and elution was conducted at
room temperature at a flow rate of 1.3mL/min, using 0.1%
formic (A phase) and acetonitrile (B phase) in the mobile
phase, under following gradient conditions: 0–3 min, 5% B;
3–7 min, 5–20% B; 7–9 min, 20% B; 9–10 min, 2–23%, B;
10–15 min, 23% B; 15–19min, 23–50% B; and 19–20min,
50–5% B, monitoring at 280 nm. Phenolic compounds were
identified by comparison with external standards (gallic and
chlorogenic acids, epigallocatechin, rutin, hyperin, quercetin,
apigenin, and kaempferol). All solutions were filtered using a
0.22-lm membrane (Millipore, Billerica, MA, USA).
Animals
Three-month-old Wistar rats of both sexes, weighing
250–300 gm, were kept under standard environmental
conditions with food and water ad libitum. All animal pro-
cedures were performed according to the Brazilian National
Health Surveillance Agency (ANVISA),
21
Organization
for Economic Cooperation and Development (OECD)
22
guidelines, and Protocol No. 035/2015, approved by the
Committee on Ethics in Animal Use, Federal University of
Rio Grande do Norte.
Activated partial thromboplastin time assay
This assay was performed in accordance with the activated
partial thromboplastin time (aPTT) information kit (CLOT
Bios Diagnostica, Sa
˜o Paulo, SP, Brazil), and the coagulation
time was measured in triplicate using a clot timer coagulometer
(Drake Electronica Commerce Ltd., Sa
˜o Paulo, Brazil).
Prothrombin time assay
The prothrombin time (PT) assay was performed accord-
ing to the manufacturer’s instructions (CLOT Bios Diag-
nostica, Sa
˜o Paulo, SP, Brazil), while the coagulation time
was measured in triplicate using a clot timer coagulometer
(Drake Electronica Commerce Ltd., Sao Paulo, Brazil).
Assay for anti-Xa activity
Regarding anti-Xa activity, the assay was performed on a
96-well microplate using the Biophen Heparin Anti-Xa kit
(Ref: 221010; HYPHEN Biomed, Paris, France) according
to the manufacturer’s instructions. Absorbance was mea-
sured at 405 nm using an Epoch microplate spectropho-
tometer (Epoch BioTek, Winooski, VT, USA).
Direct thrombin inhibition assay (anti-IIa activity)
This assessment was conducted in a 96-well microplate
using the Biophen Heparin Anti-IIa kit (ref: 221025; HY-
PHEN Biomed, Paris, France) according to the manufac-
turer’s instructions, measuring absorbance at 405 nm using a
microplate reader (Epoch BioTek, Winooski, VT, USA).
Indirect thrombin inhibition mediated
by heparin cofactor II
This parameter was determined spectrophotometrically
using standard kit assays, according to Yoon et al.,
23
mea-
suring absorbance at 405 nm using an Epoch microplate
spectrophotometer (Epoch BioTek, Winooski, VT, USA),
using a blank containing all the reagents without the test
substance.
Residual hemorrhagic effects
The CETS and AFTS compound residual hemorrhagic
effect was analyzed according to the rat topical scarification
model described by Brito et al.
24
After anesthesia with ke-
tamine and xylazine at 1:1 (v/v), an incision was performed
with a surgical blade in the distal tail portion, dipping the
tail in physiological saline to observe bleeding. In the
following stages, the tail was dipped in CETS or AFTS or
heparin solution at 100 lg/mL for 2 min and washed with
saline solution before immersing in a fresh physiological
saline solution for 40 min. Blood was quantified by the
Drabkin test and the result expressed as the hemoglobin
2LUZ ET AL.
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sum of each tube, subtracting the hemoglobin value ob-
tained before exposure to substance test.
Cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-2,
5-diphenyltetrazolium bromide assay
Mouse (3T3) and human embryonic epithelial kidney (HEK
293) cells were cultured under standard conditions in Dul-
becco’s modified Eagle’s medium (DMEM), supplemented
with 10% fetal bovine serum at 37C, 5% CO
2
, and 95%
humidity. Cells (1 ·10
4
cells/well) were cultured for 24 h in
96-well microplates to promote adhesion. Thereafter, cells
were exposed in triplicate at different CETS and AFTS con-
centrations (0.1, 1, 10, 100, and 1000 lg/mL) and incubated at
37Cfor24h.Then,100lL of 3-(4,5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide (5 mg/mL) dissolved in
DMEM was added to each well and cells incubated for 4 h.
After culture medium removal, 100 lL of dimethyl sulfoxide
was added to each well to assess cell viability at 570nm using
a microplate reader (Epoch BioTek, Winooski, VT, USA).
Acute oral toxicity. The acute oral toxicity was evalu-
ated according to ANVISA
21
and OECD
22
guidelines during
a period of 14 days. Animals randomized into five groups
(n=5) received, by gavage, doses of 500 and 2000 mg/kg, as
recommended by OECD guidelines.
22
The control group
received only distilled water. Experimental design for each
group is described as follows: Group 1: normal control rats
received only distilled water (vehicle); Group 2: received a
single dose of 500 mg/kg of CETS; Group 3: received a
single dose of 2000 mg/kg of CETS; Group 4: received a
single dose of 500 mg/kg of T. subulata ethyl acetate frac-
tion (AFTS); and Group 5: received a single dose of
2000 mg/kg of AFTS.
During the first 12 h, systematic behavioral observations
were performed (e.g., vocal tremor, piloerection, hyperac-
tivity, tremors, abdominal cramps, diarrhea, and deaths).
Behavioral parameters were evaluated in all animal experi-
mental groups. On the 14th day, animals were euthanized
with thiopental sodium (100mg/kg, i.p.), then laparotomized
for cardiac puncture blood collection and evisceration. The
liver, kidney, spleen, lung, heart, intestine, stomach, esoph-
agus, and brain were removed for macroscopic and relative
weight evaluations.
Biochemical and hematological parameters
Biochemical parameters (alanine aminotransferase, as-
partate aminotransferase, gamma-glutamyl transferase,
total protein, cholesterol, glucose, urea, creatinine, tri-
glycerides, amylase, and bilirubin) were evaluated by
commercial kits according to respective manufacturers’
instructions (Labtest kits, Lagoa Santa, MG, Brazil) on the
LabMax Plenno automated analyzer (Lagoa Santa, MG,
Brazil). Hematological parameters were determined by
ABX Micros 60 OT Equipment (ABX Diagnostics,
France). Biochemical and hematological parameters were
evaluated in all experimental groups of animals.
Statistical analyses
Results are expressed as mean SD, analyzed by a one-
way analysis of variance (ANOVA) and Tukey’s post hoc
test, with GraphPad Prism, version 5.0. Statistical signifi-
cance was considered at P<.05.
RESULTS
CETS and AFT chromatographic analyses revealed the
presence of peaks consistent with reference standards (Fig. 1),
identified by comparison with ultraviolet spectra (UV)
spectra and retention times (RT) of the two extracts and
external patterns, as depicted in Table 1. Based on this
analysis, rutin (flavonol-3-O-glycosylate) with values of
121.51 and 262.36 lg Eq/g for CETS and AFTS and values
for the apigenin-like compound (flavonoglycoside) corre-
sponding to 80.57 and 252.48 lg Eq/g for CETS and AFTS,
respectively, were identified with regard to spectral simi-
larity considering RT and UV standards.
CETS and AFTS anticoagulant activity was demonstrated
by activated partial thromboplastin and PT evaluation, such
as activated X and II factors, as well as by indirect thrombin
inhibition mediated by the heparin II cofactor.
CETS, AFTS, and heparin (standard positive control)
revealed a significant aPTT anticoagulant activity over 240 s
(negative control: 36.05 0.03 s) at 5 lg/mL, while PT >60 s
(negative control: 16.65 0.33 s) was revealed at 30 lg/mL,
as expected and displayed in Figure 2A and B, respectively.
Both CETS and AFTS inhibited clot formation through in-
trinsic and extrinsic pathways in concentrations >100 lg/mL.
Based on these data, the heparin and extract ability to
directly inhibit the activity of factors Xa and IIa (thrombin)
was evaluated. Figure 2C shows that heparin hindered factor
Xa activity in a dose-dependent concentration, requiring
1lg/mL for total factor inhibition, while 100 lg/mL of ex-
tracts completely hampered thrombin activity (Fig. 2D).
Regarding CETS and AFTS, despite their lower activities,
both extracts showed an ability to inhibit factor Xa activity in
a dose-dependent concentration, reaching a value of *40%
and 80% at *100 lg/mL to hinder thrombin activity.
To evaluate the extract’s anticoagulant action mecha-
nisms, an indirect inhibition assay was performed by heparin
cofactor II (HCII) on thrombin. Results displayed a signif-
icant extract capacity (at 100 lg/mL) to inhibit thrombin
(Fig. 2E), revealing that AFTS through HCII showed an
inhibition rate of 70%, whereas CETS showed only 30%.
Heparin’s adverse effects restrict its clinical use, such as
thrombocytopenia and hemorrhagic complications, which
interfere with the hemostatic balance. Considering the rel-
evance of these events, CETS and AFTS effects on hemo-
stasis were investigated.
Concerning the heparin assay (100 lg/mL), results dis-
played a marked hemorrhagic effect, evaluated by the high
residual bleeding level determined using the rat tail scarifi-
cation model after hemoglobin dosage in treating animals.
Regarding CETS and AFTS treatments with 100 lg/mL, both
showed anticoagulant potential with lower hemorrhagic rates,
ANTICOAGULANT POTENTIAL OF TURNERA SUBULATA 3
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FIG. 1. HPLC profile of phenolic
compounds from Turnera subulata
leaves detected at 280 nm. Profile
(A) HPLC-UV chromatogram of stan-
dard phenolic compounds, (a) gallic
acid; (b) chlorogenic acid; (c) catechin;
(d) (-)epigallocatechin gallate; (e) rutin;
(f) hyperin; (g) quercetin; (h) apigenin;
and (i) kaempferol. Profile (B) Crude
extract of T. subulata with two major-
itarian peaks: (1) related to flavonol-3-
O-glycosylate such as rutin and (2)
related to flavone glycoside such as
apigenin. Profile (C) Ethyl acetate
fraction of T. subulata with two ma-
joritarian peaks: (1) related to flavonol-
3-O-glycosylate such as rutin and (2)
related to flavone glycoside such as
apigenin. HPLC-UV, high perfor-
mance liquid chromatography coupled
to an ultraviolet detector.
4
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*50% and 10%, respectively, compared with clinical hepa-
rin (Fig. 2F).
With respect to cytotoxicity of CETS and AFTS, no cy-
totoxic effect was evidenced on normal fibroblast (3T3) and
human embryonic kidney (HEK-293) fibroblast cells
(Fig. 3A, B). No statistically significant differences were
observed relative to the negative control (DMEM). During
the in vivo toxicity assessment, no animal exhibited be-
havioral abnormalities over the trial period.
However, the biochemical parameter analysis in CETS-
and AFTS-treated animals showed statistically significant
differences with regard to glucose, triglyceride, and total
cholesterol levels. These parameters exhibited a mean de-
crease of 50% after treatment compared with the control
group (Table 2).
Regarding hematological parameters and organ weights,
no significant differences were observed relative to the
control, indicating no toxicity signs. Water and food intake
displayed no significant difference. Data regarding hema-
tological parameters and relative organ weights are attached
as Supplementary Tables S1 and S2.
DISCUSSION
Although anti-inflammatory, antidiabetic, and antiobesity
properties have been reported for the Turnera genus,
15–19
no
data regarding its anticoagulant activity have been described
so far.
Overall, plant extracts have been assessed for their ability
to inhibit blood clotting factors.
25
With regard to Passi-
floraceae species, studies have shown their potential to treat
cardiovascular diseases. The Passiflora nitida Kunth extract
showed anticoagulant activity by partially activating
thromboplastin, suggesting an inhibitory effect on intrinsic
factors of the coagulation pathway VIII, IX, XI, and XII.
26
Concerning T. subulata, both CETS and AFTS promoted
efficient inhibition of intrinsic and extrinsic pathways of the
blood coagulation cascade. This was indicated by the PT
test, also suggesting an anticoagulant effect due to thrombin
inhibition (common pathway cascade). Therefore, extracts
directly inhibited thrombin, assessed by the factor IIa inhi-
bition assay, and indirectly by HCII as a blood coagulation
action mechanism.
These parameters play a key role in coagulation inhibi-
tion. Thus, thrombin, a multifactorial enzyme, acts on the
coagulation system converting soluble plasma fibrinogen
into insoluble fibrin molecules and activating factor VIII.
This factor binds to cross-linked fibrin polymers constituting
a stable clot, amplifying the coagulation cascade with sub-
sequent factors V, VII, VIII, and XI and platelet activation
and stimulating granule release and platelet aggregation.
27
On the other hand, the long-term anticoagulant prescrip-
tion such as heparin in clinical therapy causes side effects
(e.g., bleeding, thrombocytopenia, hypersensitivity, skin
necrosis, and intestinal toxicity).
28,29
This has encouraged
the anticoagulant substance prospection to assist in clinical
treatment. However, research concerning the medicinal
plant anticoagulant effect is scarce, hence the relevance of
this study.
Thus, the inhibitory effect of CETS and AFTS on the
blood coagulation cascade may be due to the chemical
composition rich in glycosylated flavonoids, mainly rutin
and apigenin, besides other phenolic compounds. This
phytocomposition is consistent with that previously reported
for Turnera ulmifolia leaf extract, exhibiting significant
polyphenolic content, such as rutin and apigenin.
18
Studies
have demonstrated the significant anticoagulant effect of
glycosylated flavonoids.
30
Besides anticoagulant and low hemorrhagic activities,
CETS and AFTS exhibited no toxic effects in vitro using
human fibroblasts and human embryonic kidney cells. An
analogous result was reported with the Turnera diffusa
methanolic extract, exhibiting low cytotoxicity in normal
human fibroblasts.
31
Concerning the acute oral toxicity, no toxic effects were
observed regarding evaluated biochemical and hematolog-
ical parameters, especially those relating to hepatic and re-
nal functions. There are no data regarding the T. subulata
toxicity potential. However, the low acute oral toxicity level
of T. diffusa Willd corroborates the results of the present
study since no behavioral change, mortality, renal, and he-
patic histopathology, as well as biochemical, parameters
were observed.
32
Although AFTS displayed side effects
lower than CETS due to higher polyphenol content, both
extracts can be considered safe for clinical use.
33,34
Al-
though no acute toxic effects have been observed, anato-
mopathological analyses of different tissues or organs such
as the liver or kidney are required to corroborate the absence
of damage to the liver and/or kidneys. This evaluation will
be carried out in the continuation of the present study.
A relevant aspect regarding the biochemical parameters
evaluated in this study was the decrease in glucose, tri-
glyceride, and total cholesterol values after the CETS and
AFTS treatments. These results suggest hypoglycemic
and hypolipidemic properties, which indicate the phy-
totherapeutic potential of these extracts for cardiovascu-
lar disease treatment. These effects have been reported in
Table 1. Quantification Parameters of Nine Phenolic
Compounds Identified by Comparison Between UV Spectra
and Retention Times of the Extracts and External
Standards Based on the Chromatographic Method
Compound
Retention
times
(min)
Linear
range
(lg/mL)
UV
(nm) R
2
Gallic acid 1.71 1.5–50 271 0.9945
Chlorogenic acid 6.47 1.5–50 326 0.9973
Catechin 7.19 1.5–50 278 0.9993
(-) Epigallocatechin
gallate
8.18 1.5–50 275 0.9989
Rutin 9.20 1.81–100 275–354 0.9976
Hyperin 9.24 1.81–100 256–354 0.9995
Quercetin 16.03 1.5–50 255–371 0.9991
Apigenin 18.51 1.5–50 266–337 0.9989
Kaempferol 18.85 1.5–50 263–367 0.9984
UV, ultraviolet spectra.
ANTICOAGULANT POTENTIAL OF TURNERA SUBULATA 5
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FIG. 2. Anticoagulant activity of CETS, AFTS, and heparin by activated partial thromboplastin time assay (A), prothrombin time assay (B),
inhibition of factor Xa activity (C), inhibition of thrombin activity (D), indirect thrombin inhibition activity mediated by heparin cofactor II (E),
and bleeding activity of CETS, AFTS, and porcine intestinal mucosa heparin (100 lg/mL) applied topically (F). AFTS, acetate fraction of
T. subulate; CETS, T. subulata crude extract.
6
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several Passifloraceae species, especially in the Turneraceae
genus,
16,35–38
for example, in T. diffusa
35
and T. ulmifolia,
36
whose extracts were evaluated in animal models.
Overall, results regarding CETS and AFTS anticoagulant,
nontoxic, and low hemorrhagic effects are due to their sig-
nificant polyphenolic compound content. These compounds
are associated with several pharmacological activities
widely described in the literature.
39,40
In conclusion, T. subulata CETS and AFTS results dis-
played anticoagulant activity, inhibiting intrinsic and ex-
trinsic pathways in the blood coagulation cascade. This
suggests direct and indirect thrombin inhibition as the main
action mechanism. Moreover, extracts showed low hem-
orrhagic and toxic effects in vitro and in vivo, while the
biochemical parameters showed possible hypoglycemic
and hypolipidemic activities. These biological effects can
be attributed to the significant polyphenol content. Overall,
experimental results are promising due to the T. subulata
therapeutic potential to develop herbal formulations to
assist in anticoagulant therapy, although further studies are
required, considering that this is the first report evaluating
the anticoagulant capacity of this plant.
ACKNOWLEDGMENTS
This research was supported by the Conselho Nacional de
Desenvolvimento de Cientı
´fico e Tecnolo
´gico (CNPq)
(Protocol No.478652/2010-0) and Banco do Nordeste
(Protocol No.912011) grants. The authors would like to
thank Coordenac¸a
˜o de Aperfeic¸oamento de Pessoal de
FIG. 3. Cell viability (cytotoxicity effects) of CETS and AFTS on mouse fibroblast cells (3T3) (A) and epithelial embryonic human kidney cells
(HEK 293) (B), measured by MTT assays. Culture medium DMEM was used as a negative control of cytotoxicity. Comparisons between groups
were analyzed with an ANOVA and Tukey’s post hoc test. ANOVA, analysis of variance; DMEM, Dulbecco’s modified Eagle’s medium; MTT,
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide.
Table 2. Biochemical Parameters of Rats After 14 Days of Treatment with CETS and AFTS
Biochemical parameters Control 2000 mg/kg CETS 500 mg/kg CETS 2000 mg/kg AFTS 500 mg/kg AFTS 2000 mg/kg
Glic (mg/dL) 122 21.9 60 20.1* 61.4 21.6* 60.1 19.2* 62 21.4*
Trig (mg/dL) 53 5.45 25 3.89* 28.6 4.38* 25 2.32* 28.7 4.34*
Col (mg/dL) 54.5 6.05 24.5 0.24* 22 0.19* 25.5 0.19* 21.5 0.10*
ALT (U/L) 97 4.24 86.1 2.30 89 2.20 89.9 4.34 97 5.29
AST (U/L) 250 23.3 238 25.3 249 15.3 228.5 24.8 240.3 14.1
c-GT (U/L) 12 1.4 11 1.5 10.66 0.05 12 0.23 11.5 0.04
TB (mg/dL) 0.99 0.01 0.90 0.01 0.93 0.01 0.90 0.01 0.99 0.02
DB (mg/dL) 0.99 0.07 0.98 0.05 0.97 0.09 0.99 0.05 0.110 0.1
IB (mg/dL) 0.28 0.01 0.13 0.03 0.12 0.09 0.15 0.03 0.15 0.08
Urea (mg/dL) 54.5 7.7 54.5 7.5 57.3 7.04 55.9 7.5 58.3 7.75
Cret (mg/dL) 0.65 0.07 0.65 0.07 0.7 0.03 0.59 0.19 0.70 0.07
TP (g/dL) 6.6 0.1 6.2 0.5 6.3 0.64 6.1 0.5 6.9 0.6
ALB (g/dL) 3.1 0.42 3.9 0.42 2.9 0.34 3.1 0.42 3 0.12
Glo (g/dL) 3.5 0.42 3.8 0.35 4.3 0.54 3.1 0.32 4.82 0.54
Ami (U/L) 900 3.53 900 3.44 903 2.09 902.5 3.43 913.8 2.07
Results are expressed as mean SD (n=5); control group treated with vehicle (distilled water). Comparisons between groups were analyzed with an ANOVA and
Tukey’s post hoc test.
*P<.05 compared with the control group.
AFTS, acetate fraction of T. subulate; ALT, alanine aminotransferase enzymes; ALB, albumin; Ami, amylase; ANOVA, analysis of variance; AST, aspartate
aminotransferase; CETS, T. subulata crude extract; Col, cholesterol; Cret, creatinine; DB, direct bilirubin; c-GT, gamma-glutamyl transferase; Glo, globulin; Gli,
glucose; IB, indirect bilirubin; TB, total bilirubin; TP, total proteins; Trig, triglycerides.
ANTICOAGULANT POTENTIAL OF TURNERA SUBULATA 7
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´vel Superior (CAPES) and CNPq for providing a post-
graduation fellowship and the Department of Biochemistry
(UFRN) for cell culture technical assistance.
AUTHOR DISCLOSURE STATEMENT
No competing financial interests exist.
SUPPLEMENTARY MATERIAL
Supplementary Table S1
Supplementary Table S2
REFERENCES
1. World Health Organization: Cardiovascular Diseases (CVDs).
Geneva, Switzerland, 2016. www.who.int/cardiovascular_diseases/
en/ (accessed August 1, 2018).
2. Stewart J, Manmathan G, Wilkinson P: Primary prevention of
cardiovascular disease: A review of contemporary guidance and
literature. JRSM Cardiovasc Dis 2017;6:1–9.
3. Weitz JI, Eikelboom JM, Samama MM: New antithrombotic
drugs: Antithrombotic therapy and prevention of thrombosis, 9th
ed: American College of Chest Physicians evidence-based clin-
ical practice guidelines. Chest 2012;141:120–151.
4. Bauer KA: New anticoagulants: Anti IIa vs Anti Xa—Is one
better? J Thromb Thrombolys 2006;21:67–72.
5. Kaeron C, et al.: Antithrombotic therapy for VTE disease: An-
tithrombotic therapy and prevention of thrombosis, 9th ed:
American College of Chest Physicians evidence-based clinical
practice guidelines. Chest 2012;141:419–494.
6. Revel-Vilk S, et al.: Quantification of bleeding symtoms in a
national registry of patients with inherited platelet disorders.
Blood Cells Mol Dis 2017;67:59–62.
7. Zhu Y, et al.: Structure-guided creation of AcAP5-derived and
platelet targeted factor Xa inhibitors. Biochem Pharmacol 2015;
95:253–262.
8. Liu MM, Zheng MMY, Liu DMG: Safety of recanalization
therapy in patients with acute ischemic stroke under anticoagu-
lant: A systematic review and meta-analysis. J Stroke Cere-
brovasc Dis 2018;27:2296–2305.
9. Zbidi H, et al.: Olive tree wood phenolic compounds with human
platelet antiaggregant properties. Blood Cells Mol Dis 2018;42:
279–285.
10. Pour MA, et al.: Evaluating the anticoagulant effect of medicinal
plants in vitro by cheminformatics methods. J Herbal Med 2016;
6:128–136.
11. Akram M, Rashid A: Anti-coagulant activity of plants: Mini
review. J Thromb Thrombolysis 2017;44:406–411.
12. Arbo MM: Turneraceae (Turnera family). In: Flowering Plants
of the Neotropics (Smith N, Mori AS, Henderson A, Stevenson
DW, eds.). Princeton University, New Jersey, 2004, pp. 380–382.
13. Galvez J, et al.: Intestinal antiinflammatory activity of a lyoph-
ilized infusion of Turnera ulmifolia in TNBS rat colitis. Fito-
terapia 2006;77:515–520.
14. Souza NC, et al.:Turnera subulata anti-inflammatory properties
in lipopolyssacharides-stimulated RAW 264.7 macrophages.
J Med Food 2016;19:191–199.
15. Alarco
´n-aguilara FJ, et al.: Study of the anti-hyperglicemic effect of
plants used as antidiabetics. J Ethnopharmacol 1998;61:101–61110.
16. Santos KKA, et al.: Anti-candida activity of Mentha arvensis and
Turnera ulmifolia.J Med Food 2012;15:322–324.
17. Brito NJN, et al.: Antioxidant activity and protective effect of
Turnera ulmifolia Linn. var. elegans against carbono tetrachloride-
induced oxidative damage in rats. Food Chem Toxicol 2012;50:
4340–4347.
18. Gadioli IL, et al.: A systematic review on phenolic com-
pounds in Passiflora plants: Exploring biodiversity for food,
nutrition, and popular medicine. Crit Rev Food Sci Nutr 2018;
58:785–807.
19. Kumar S, Taneja R, Sharma A: The genus Turnera: A review
update. Pharm Biol 2005;43:383–391.
20. George
´S, Brat P, Alter P, Amiot MJ: Rapid determination of
polyphenols and vitamin C in plant-derived products. J Agri
Food Chem 2005;531370–531378.
21. National Health Surveillance Agency: Ministry of Health Re-
solution No. RE 90/2004. Standards for toxicological studies of
herbal products, Official Gazette of the Federative Republic of
Brazil, Executive Branch. Brası
´lia, DF, 2004.
22. Organization for Economic Cooperation and Development:
Guidelines for the Testing of Chemicals: Acute and Toxicity–
Acute Toxic Class Method. OECD Guideline, Paris, 423,
2001.
23. Yoon SJ, et al.: The medicinal plant Porana volubilis contains
polysaccharides with anticoagulant activity mediated by heparin
cofactor II. Thromb Res 2002;10651–10658.
24. Brito AS, et al.: A non-hemorrhagic hybrid heparin/heparan
sulfate with anticoagulant potential. Carbohydr Polym 2014;99:
372–378.
25. Chaves DSA, et al.: Secondary metabolites from vegetal origin:
A potential source of antithrombotic drugs. Quim Nova 2010;33:
1980–1986.
26. Carvalho MJ, et al.: Pharmacognostic study and in vitro activity
on blood coagulation and platelet aggregation of leaves of Pas-
siflora nitida Kunth (Passifloraceae). Acta Amazonica 2010;40:
199–206.
27. Periayah MH, Halim AS, Saad AZM: Mechanism ac-
tion of platelets and crucial blood coagulation pathways in
hemostasis. Int J Hematol Oncol Stem Cell Res 2017;11:
319–327.
28. Wang SV, et al.: Prediction of rates of thromboembolic and
major bleeding outcomes with dabigatran or warfarin among
patients with atrial fibrillation: New initiator cohort study. BMJ
2016:353:1–10.
29. Okishige K, et al.: Comparative study of hemorrhagic and is-
chemic complications among anticoagulants in patients under-
going cryoballoon ablation for atrial fibrillation. J Cardiol 2017;
69:11–15.
30. Liu J, et al.: Isolation, structural characterization and bioactiv-
ities of naturally occurring polysaccharide-polyphenolic conju-
gates from medicinal plants—A review. Int J Bio Macromol
2018;107:2242–2250.
31. Avelino-Flores MDC, et al.: Cytotoxic activity of the methanolic
extract of Turnera diffusa Willd on breast cancer cells. J Med
Food 2015;18:299–305.
32. Taha MME, et al.: Gastroptotective activities of Turnera diffusa
Willd. Ex Schult. Revisited: Role of arbutin. J Ethnopharmacol
2012;141:273–281.
33. Palhares RM, et al.: Medicinal plants recommended by
the World Health Organization: DNA barcade identification
8LUZ ET AL.
Downloaded by University of Florida E-journal package from www.liebertpub.com at 03/24/19. For personal use only.
associates with chemical analyses guarantees their quality.
PLos One 2015;10:1–29.
34. Senes-Lopes TF, et al.: Genotoxicity of Turnera subulata and
Spondias mombin ·Spondias tuberosa extracts from Brazilian
caatinga biome. J Med Food 2018;21:372–379.
35. Naranjo AP, et al.: Acute hypoglycemic and antidiabetic effect of
teuhetenone A isolated from Turnera diffusa.Molecules 2017;22:
599–612.
36. Gonza
´lez ALU, et al.: Natural vaiability of essential oil and
antioxidants in the medicinal plant Turnera diffusa.Asian Pac J
Trop Med 2017;10:121–125.
37. Dutra RC, Campos MM, Santos ARS, Calixto JB: Medicinal
plants in Brazil: Pharmacological studies, drug discovery, chal-
lenges and perpectives. Pharmacol Res 2016;112:4–29.
38. Sudasingle HP, Peiris DC: Hypoglycemic and hypolipidemic
activity of aqueous leaf extract of Passiflora suberosa L. Peer J
2018;6:e4389.
39. Rasouli H, Farzei MH, Khodarahmi R: Polyphenols and their
benefits: A review. Int J Food Prop 2017;20:1700–1741.
40. Cutrim CS, Corte
´z MAS: A review on polyphenols: Classifica-
tion, beneficial effects and their application in dairy products. Int
J Dairy Technol 2018;71:564–578.
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