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Anticoagulant activities of curcumin and its derivative

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Curcumin, a polyphenol responsible for the yellow color of the curry spice turmeric, possesses antiinflammatory, antiproliferative and antiangiogenic activities. However, anticoagulant activities of curcumin have not been studied. Here, the anticoagulant properties of curcumin and its derivative (bisdemethoxycurcumin, BDMC) were determined by monitoring activated partial thromboplastin time (aPTT), prothrombin time (PT) as well as cell-based thrombin and activated factor X (FXa) generation activities. Data showed that curcumin and BDMC prolonged aPTT and PT significantly and inhibited thrombin and FXa activities. They inhibited the generation of thrombin or FXa. In accordance with these anticoagulant activities, curcumin and BDMC showed anticoagulant effect in vivo. Surprisingly, these anticoagulant effects of curcumin were better than those of BDMC indicating that methoxy group in curcumin positively regulated anticoagulant function of curcumin. Therefore, these results suggest that curcumin and BDMC possess antithrombotic activities and daily consumption of the curry spice turmeric might help maintain anticoagulant status.
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BMB
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orts
221http://bmbreports.org BMB reports
*Corresponding author. Tel: +82-53-950-8570; Fax: +82-53-950-
8557; E-mail: baejs@knu.ac.kr
#First two authors contributed equally to this work.
http://dx.doi.org/10.5483/BMBRep.2012.45.4.221
Received 9 August 2011, Revised 28 September 2011,
Accepted 11 October 2011
Keywo rds : aPTT, Curcumin, HUVECs, PT
Anticoagulant activities of curcumin and its derivative
Dong-Chan Kim1,#, Sae-Kwang Ku2,# & Jong-Sup Bae3,*
1Laboratory of Microvascular Circulation Research, NEUORNEX Inc. Daegu 711-823, 2Department of Anatomy and Histology, College of
Oriental Medicine, Daegu Haany University, Gyeongsan 712-715, 3College of Pharmacy, Research Institute of Pharmaceutical Sciences,
Kyungpook National University, Daegu 702-701, Korea
Curcumin, a polyphenol responsible for the yellow color of
the curry spice tu rmeric, p os sesses anti inflammatory, anti-
proliferative and antiangiogenic activities. However, anti-
coagulant activities of curcumin have not been studied. Here,
the anticoagulant properties of curcumin and its derivative
(bisdemethoxycurcumin, BDMC) were determined by monitor-
ing activated partial thromboplastin time (aPTT), prothrombin
time (PT) as well as cell- base d thrombi n and activated factor X
(FXa) generation activities. Data showed that curcumin and
BDMC prolonged aPTT and PT significantly and inhibited
thrombin and FXa activities. They inhibited the generation of
thrombin or FXa. In accordance with these anticoagulant activ-
ities, curcumin and BDMC showed anticoagulant effect in
vivo. Surprisingly, these anticoagulant effects of curcumin
were be tter th an those of BDMC indi catin g that methoxy
group in curcumin positively regulated anticoagulant function
of curcumin. Therefore, these results suggest that curcumin
and BDMC pos sess antithrombotic activities and dai ly con-
sumption of the curry spice turmeric might help mai ntain anti-
coagulant status. [BMB reports 2012; 45(4): 221-226]
INTRODUCTION
The key of the blood clotting pathway is the production of
thrombin which is required for the conversion of fibrinogen to
fibrin (1, 2). Thrombin resides in the cell in an inactive form,
called prothrombin, and is activated by the coagulation casca-
devia formation of a complex called the prothrombin activator
complex (1-5). The formation of the prothrombin activator com-
plex occurs by two different pathways: the intrinsic pro-
thrombin activation pathway and the extrinsic prothrombin ac-
tivation pathway. Though the ultimate goal of both the path-
ways is the generation of the prothrombin activator complex,
alternate routes are used, each giving rise to a different form of
the prothrombin activator (1-5). In the extrinsic pathway, pro-
thrombin activator complex consists of activated factor X (FXa),
tissue factor (TF), activated factor VII (FVIIa) and the cofactor
activated factor V (FVa) (1-5). This complex, specifically FXa,
along with the cofactor FVa, then converts prothrombin to ac-
tive thrombin. Fibrin forms a mesh within the platelet aggregate
to stabilize clots (1-5). In contrast, in the intrinsic pathway, pro-
thrombin activator complex consists of FXa, FVa, activated fac-
tor VIII (FVIIIa) and phopspholipid (PL) (1-5). The clotting time
assay measures the lag time of thrombin generation (6) and the
activated partial thromboplastin time (aPTT) is a performance
indicator measuring the efficacy of both the contact activation
pathway and the common coagulation pathways (6). Further,
the prothrombin time (PT) is measure of the extrinsic pathway
of coagulation (7, 8).
 The rhizome of Curcuma longa has been used in indigenous
medicine for the treatment of inflammatory disorders and its me-
dicinal activity has been known since ancient times. Turmeric
derived from the rhizome has been widely used by the people in
the Middle East for centuries as a food component (9, 10). The
use of turmeric extract or turmeric oil as a spice and household
remedy has been known to be safe for centuries. Bhide, et al. al-
so revealed the safety and tolerance of turmeric through human
clinical trials (11). In many previous studies, extracts prepared
from Curcuma longa have been used as antiinflammatory agents
to treat gas, colic, toothaches, chest pains, menstrual difficulties,
stomach and liver ailmetns (9, 12, 13). Polyphenolic phytochem -
icals are common in the diet and have been suggested to have a
wide range of beneficial health effects and the polyphenolic
compounds in turmeric are responsible for a number of its bene-
ficial health effects (14, 15). Turmeric contains three major poly-
phenolic analogues. The majority is curcumin and the com-
pounds in smaller amounts are demethoxycurcumin, and bisde-
methoxycurcumin (BDMC) (16, 17). Recent studies indicate that
dietary administration of curcumin may have beneficial effects in
conditions such as cancer (18), Alzheimer’s disease (19) and
cystic fibrosis (20). With regard to mode of action, curcumin ex-
hibits a diverse array of metabolic, cellular and molecular activi-
ties. Although curcumin analogues exhibit activities very similar
to curcumin, their potencies compared to curcumin have not
been clearly established. In most systems, curcumin is found to
be most potent (21, 22) and in some systems, BDMC was found
Anticoagulant activities of curcumin
Dong-Chan Kim, et al.
222 BMB reports http://bmbreports.org
In vitro coagulant assay
Sample Dose aPTT (s) PT (s)
Control
Curcumin
Heparin
Saline
0.1 μM
0.5 μM
1 μM
5 μM
10 μM
20 μM
50 μM
36.2 ± 1.2
37.2 ± 1.3
48.5 ± 1.4b
52.6 ± 1.8c
65.3 ± 1.5c
77.5 ± 2.1c
91.8 ± 1.5c
119.8 ± 0.9c
1.5 (μg/ml)
300c
17.5 ± 0.4
17.4 ± 0.3
18.2 ± 0.7
19.8 ± 0.5b
21.6 ± 0.6b
27.5 ± 0.5c
31.8 ± 0.4c
35.2 ± 0.4c
15 (μg/ml)
61.5 ± 0.5c
In vivo bleeding time
Sample Dose Tail Bleeding time (s) n
Control
Curcumin
Heparin
Saline
100 mg/kg
50 mg/kg
54.2 ± 8
102 ± 2c
158.6 ± 4c
3
3
3
aEach value represents the means ± SD (n = 5). bP 0.05 as com-
pared to control. cP 0.01 as compared to control
Table 1. Anticoagulant activity of curcumina
In vitro coagulant assay
Sample Dose aPTT (s) PT (s)
Control
BDMC
Heparin
Saline
0.1 μM
0.5 μM
1 μM
5 μM
10 μM
20 μM
50 μM
35.8 ± 1.3
38.9 ± 0.8
41.6 ± 1.5b
48.5 ± 2.14b
68.5 ± 1.2c
70.5 ± 1.8c
87.6 ± 1.5c
98.6 ± 1.4c
1.5 (μg/ml)
300c
17.5 ± 0.4
17.5 ± 0.5
17.9 ± 0.6
18.2 ± 0.5b
19.2 ± 0.7c
20.9 ± 0.4c
25.4 ± 0.3c
29.8 ± 0.5c
15 (μg/ml)
61.5 ± 0.5c
In vivo bleeding time
Sample Dose Tail Bleeding time (s) n
Control
BDMC
Heparin
Saline
100 mg/kg
50 mg/kg
54.2 ± 8
82 ± 2b
158.6 ± 4c
3
3
3
aEach value represents the means ± SD (n = 5). bP 0.05 as
compared to control. cP 0.01 as compared to control
Table 2. Anticoagulant activity of BDMCa
to exhibit different (in some cases, more potent) activities (23-25).
There is an increasing demand for comparison study between
curcumin and BDMC, due to the discovery of their new bio-
logical activities (21, 26, 27). Identification of novel biological
activities of curcumin and its analogues is of interest both pre-
clinically and clinically. Additionally, anticoagulant activities of
curcumin have not been well studied. Herein, the anticoagulant
properties of curcumin and its derivative, BDMC on the gen-
eration of FXa and thrombin as well as the regulation of clotting
time (PT and aPTT) were determined.
RESULTS
Effects of curcumin and BDMC on aPTT and PT
The anticoagulant properties of curcumin and BDMC were
tested in aPTT and PT assays using human plasma and are
summarized in Table 1 and 2. Although the anticoagulant ac-
tivities of curcumin and BDMC were weaker than those of
heparin, aPTT and PT were significantly prolonged by curcu-
min or BDMC at concentrations at or greater than 5 μM.
Prolongation of aPTT suggests inhibition of the intrinsic and/or
the common pathway while prolonged PT indicates that curcu-
min and BDMC could also inhibit the extrinsic pathway of
coagulation. To confirm these in vitro data, in vivo tail bleed-
ing time was determined. As shown in Table 1 and 2, tail
bleeding time was significantly prolonged by curcumin or
BDMC with respect to the control. Surprisingly, effects of cur-
cumin on the clotting time were better than that of BDMC sug-
gesting that methoxy group in curcumin positively regulates
the anticoagulant function of curcumin.
Effects of curcumin and BDMC on inactivation of thrombin or
FXa
To elucidate the inhibitory mechanism of curcumin and
BDMC on coagulation time, their inhibitory effect on thrombin
and FXa activities was measured using chromogenic substrates
in the absence or presence of antithrombin III (AT III). In the
absence of AT III, the amidolytic activity of thrombin was in-
hibited by curcumin and BDMC in a dose-dependent manner,
showing that the anticoagulant directly inhibited thrombin
activity. However, in the presence of AT III, thrombin activity
was essentially unchanged (Fig. 1A, B). Thus, AT III was un-
able to potentiate the activity of curcumin or BDMC. Further,
the effects of curcumin and BDMC on FXa activity in the ab-
sence or presence of AT III were also investigated. The anti-
coagulant showed direct inhibitory effects on FXa activities at
high concentrations, and the inhibitory effect of AT III was not
changed by curcumin or BDMC (Fig. 1C, D). These results
were consistent with the antithrombin assay. Therefore, these
results suggested that the antithrombotic mechanism of curcu-
min and BDMC appears to be due to inhibition of fibrin poly-
merization and/or the intrinsic/extrinsic pathway without po-
tentiation by AT III. Furthermore, the methoxy group in curcu-
min positively regulates the anticoagulant effects on the in-
hibition of thrombin or FXa activity because the anticoagulant
effects of curcumin were better than those of BDMC.
Anticoagulant activities of curcumin
Dong-Chan Kim, et al.
223http://bmbreports.org BMB reports
Fig. 1. Effect of curcumin and BDMC on the inactivation o
f
thrombin and factor Xa. Inhibition of thrombin (Th) in the ab-
sence of antithrombin III () or in the presence of antithrombin
III () by curcumin (A) or DBMC (B) was monitored by a chro-
mogenic assay as described in “Materials and Methods”. Inhibition
of factor Xa (FXa) in the absence of antithrombin III () or in
the presence of antithrombin III () by curcumin (C) or DBMC
(D) was monitored by chromogenic assay as described in
“Materials and Methods”. *P 0.01 as compared to 0.
Fig. 2. Inhibition of thrombin and FXa generation by curcumin
and BDMC in HUVECs. (A) FVa (100 pM) and FXa (1 nM) were
preincubated on HUVECs monolayers for 10 min with indicated
concentrations of curcumin () or BCMC (). Prothrombin was
added to a final concentration of 1 μM and the amounts of pro -
thrombin activated were determined at 30 min as described in
“Materials and Methods”. (B) HUVECs were preincubated with in-
dicated concentrations of curcumin () or BCMC () for 10 min.
TNF-α (10 ng/ml for 6 h) stimulated HUVECs were incubated with
FVIIa (10 nM) and FX (175 nM) in the absence or presence o
f
anti-TF IgG (25 μg/ml) and FXa generation was then determined
as described in “Materials and Methods”. (C) HUVECs were cul-
tured with curcumin () or BCMC () in the absence or pres -
ence of TNF-α (10 ng/ml) for 18 h and PAI-1 concentration in the
culture mediums was examined as described in “Materials and
Methods”. (D) the same as C except that cells were preincubated
with curcumin () or BCMC () and cultured with SP600125 (2
μM), emodin (2 μg/ml), and PD98059 (10 μM) in presence o
f
TNF-α (10 ng/ml) for 18 h. *P 0.05 as compared to 0 (A) or
TNF-α alone (B, C) or to (+) control (D).
Effects of curcumin and BDMC on the generation of thrombin
and FXa
Sugo et al. reported that endothelial cells are able to support
prothrombin activation by FXa (28). Preincubation of FVa and
FXa in the presence of CaCl2 with HUVECs before addition of
prothrombin resulted in thrombin generation (Fig. 2A). The ef-
fect of curcumin and BDMC on thrombin generation showed
that curcumin and BDMC inhibited thrombin activation of pro-
thrombin dose- dependently (Fig. 2A). Rao et al. showed that
the endothelium provides the functional equivalent of procoa-
gulant phospholipids and supports FX activation (29) and that
in TNF-α stimulated HUVECs, FVIIa could activate FX, which
was completely dependent on TF expression (30). Thus, it is
likely that the endothelium can provide support for FVIIa acti-
vation of FX. If so, it would be of interest to investigate the ef-
fect of curcumin and BDMC on FVIIa activation of FX.
HUVECs were stimulated with TNF-α to induce TF expression.
As shown in Fig. 2B, the rate of FX activation by FVIIa was
100-fold higher in stimulated HUVECs (53.3 ± 5 nM) com-
pared with non-stimulated HUVECs (0.54 ± 0.2 nM), which
was completely attenuated by anti-TF IgG (4.8 ± 0.4 nM).
Moreover, preincubation with curcumin or BDMC dose-de-
pendently inhibited FVIIa activation of FX (Fig. 2B). Therefore,
these results suggested that curcumin could inhibit the gen-
eration of thrombin or FXa and the methoxy group in curcu-
min positively regulated these functions of curcumin.
 Because plasminogen activator inhibitor type 1 (PAI-1) de-
termines fibrinolytic activity (1), the effect of TNF-α and curcu-
min on PAI-1 secretion from HUVECs were investigated. As
shown in Fig. 2C, curcumin dose dependently inhibited TNF-α
induced PAI-1 secretion from HUVECs. To define the molec-
ular targets of curcumin in the signal transduction pathways
leading to TNF-α induced PAI-1 expression, we investigated
the effects of three signal transduction inhibitors, emodin (a
NF-κB inhibitor), PD98059 (an extracellular signal regulated
kinase, ERK, inhibitor), and SP600125 (a c-Jun N-terminal kin-
ase, JNK, inhibitor) on TNF-α induced PAI-1 expression in the
presence or absence of curcumin. Experiments performed
showed that neither SP600125 nor emodin showed any addi-
tional inhibitory effects in the presence of curcumin (Fig. 2D).
However, the inhibitory effects of PD98059 were essentially
additive with those of curcumin (Fig. 2D).
DISCUSSION
The vascular endothelium provides a number of important
functions in order to maintain adequate blood supply to vital
organs. These functions include prevention of coagulation,
regulation of vascular tonus, orchestration of the migration of
blood cells by the expression of adhesion molecules and regu-
Anticoagulant activities of curcumin
Dong-Chan Kim, et al.
224 BMB reports http://bmbreports.org
lation of vasopermeability (31). Among these, regulation of he-
mostatic activity was regulated through a balance of pro- and
anticoagulant properties (1). Impaired endothelial function
causes thrombus-related complications including myocardial
infarction, stroke and thromboembolism (32). In this study, we
presented curcumin as a potent anticoagulant by inhibitiing
thrombin or FXa. The anticoagulant activity of curcumin was
evidenced by the prolongation of the clotting time in plas-
ma-based PT and APTT assays. Additionally, the inhibitory ef-
fects by curcumin on FXa generation and further thrombin
generation support the anticoagulant activities of curcumin.
 It is well known that FXa has no effect on platelet activation,
however, once it is assembled into the prothrombinase com-
plex, it triggers enormous amounts of thrombin (2, 5). Throm-
bin is the final enzyme in the blood clotting cascade respon-
sible for clot formation and platelet activation (2, 5). Based on
the results that curcumin could inhibit generation of FXa and
thrombin, the anticoagulant activity of curcumin was initiated
from the inhibition of the penultimate and final enzyme in the
blood clotting cascade.
 TNF-α has been known to activate JNK, NF-κB, and ERK in
human endothelial cells (33-35). Here, we used a JNK
(SP600125), NF-κB (emodin), and ERK (PD98059) inhibitors to
define the molecular target of curcumin. We observed that
PD98059, but not emodin or SP600125, was additive to the in-
hibitory effects of curcumin on TNF-α induced PAI-1 secretion.
These results suggest that the NF-κB and JNK pathway are in-
volved in curcumin mediated inhibition of TNF-α induced
PAI-1 expression in HUVECs. Thus, these results seem to in-
dicate that curcumin decreases PAI-1 levels via inhibition of the
NF-κB and JNK pathways.
 Noting that the effects of curcumin on the anticoagulant ac-
tivity was better than BDMC, it suggests that the ortho-methoxy
group in curcumin positively regulates anticoagulant functions
of curcumin. In a previous report, curcumin and BDMC had
different redox properties due to the presence of the ortho-me-
thoxy group in position 3 of the phenyl moiety in curcumin.
(36) While curcumin has two symmetric ortho-methoxy phe-
nols linked through the a,b-unsaturated b-diketone moiety,
BDMC, which is also symmetric, is deficient in the two or-
tho-methoxy substitutions. Although curcumin and bisdeme-
thoxycurcumin differ in their chemical structures only with re-
gard to the ortho-methoxy substitution, they exhibit signifi-
cantly different antioxidant, antitumor, and antiinflammatory
activities. The hydrogen bonding interaction between the phe-
nolic OH and the ortho-methoxy groups in curcumin markedly
influences the O-H bond energy and H-atom abstraction by
free radicals, thus making it a better free radical scavenger than
BDMC (37). In another investigation, the ortho-methoxy- defi-
cient BDMC was a more potent ROS inducer and the ortho-me-
thoxy substituted curcumin was a more potent suppressor of
NF-kB activation (38). According to our results, the ortho-me-
thoxy group in curcumin is important for the anticoagulant
effect. Thus, we can postulate that the anticoagulant activities
of curcumin could be mainly caused by interaction of the target
molecules with the ortho-methoxy group.
 The significant progress made in understanding the role of
FXa and thrombin in various thrombotic disease states has
clearly demonstrated potential therapeutic benefits of blocking
these key enzymes in the blood coagulation cascade (39). A
potent and selective small molecule FXa or thrombin inhibitor
has the potential to offer substantial therapeutic benefits (39).
Curcumin exhibits the potency and selectivity required for such
a candidate and is currently undergoing additional evaluations.
 In conclusion, this study showed that curcumin inhibited the
extrinsic and intrinsic pathways of blood coagulation by in-
hibiting FXa and thrombin generation in HUVECs. These re-
sults adds to previous work and may be helpful for the rational
design of pharmacological strategies for treating or preventing
vascular diseases via regulation of thrombin generation.
MATERIALS AND METHODS
Reagents
Curcumin (product catalog #: C2302) and bisdemethoxycurcu-
min (product catalog #: B3347) were purchased from TCI
Korea (Tokyo Chemical Industry Co., Ltd. Seoul, South Korea).
TNF-α, JNK inhibitor (SP600125), NF-κB inhibitor (emodin),
and ERK inhibitor (PD98059) were purchased from R&D
Systems (Minneapolis, MN). Anti-tissue factor antibody was
purchased from Santa Cruz Biologics (Santa Cruz, CA). Factor
V, Vll, Vlla, FX, FXa, antithrombin III (AT III), prothrombin and
thrombin were obtained from Haematologic Technologies
(Essex Junction, VT, USA). aPTT assay reagent and PT reagents
were purchased from Fisher Diagnostics (Middletown, Virgi-
nia, USA). Chromogenic substrates S-2222, and S-2238 were
from Chromogenix AB (Sweden).
Anticoagulation assay
Determination of aPTT and PT were performed according to
the manufacture’s specifications using Thrombotimer (Behnk
Elektronik, Germany). In brief, citrated normal human plasma
(90 μl) was mixed with 10 μl of curcumin or BDMC and in-
cubated for 1 min at 37°C. Then, aPTT assay reagent (100 μl)
was added to the mixture and incubated for 1 min at 37°C.
Thereafter, 20 mM CaCl2 (100 μl) was added and the clotting
time was recorded. For the PT assay, citrated normal human
plasma (90 μl) was mixed with 10 μl of a curcumin or BDMC
stock and incubated for 1 min at 37oC. Then, PT assay reagent
(200 μl), preincubated for 10 min at 37oC, was added and the
clotting time was recorded.
Cell culture
Primary HUVECs were obtained from Cambrex Bio Science
(Charles City, IA) and maintained as described before (40).
Briefly, cells were cultured to confluency at 37oC at 5% CO2
in EBM-2 basal media supplemented with growth supplements
(Cambrex Bio Science).
Anticoagulant activities of curcumin
Dong-Chan Kim, et al.
225http://bmbreports.org BMB reports
Factor Xa generation on the surface of HUVECs
HUVECs were preincubated with indicated concentrations of
curcumin or BDMC for 10 min. TNF-α (10 ng/ml for 6 h in
serum-free medium) stimulated confluent monolayers of
HUVECs in a 96-well culture plate were incubated with FVIIa
(10 nM) in buffer B for 5 min at 37°C in presence or absence
of anti-TF IgG (25 μg/ml). FX (175 nM) was then added to the
cells (final reaction mixture volume, 100 μl) and incubated for
15 min. The reaction was stopped by adding buffer A contain-
ing 10 mM EDTA and the amount of FXa generated in the re-
action period was measured by using a chromogenic substrate,
and the change in absorbance at 405 nm was monitored in a
microplate reader for 2 min. The initial rate of color develop-
ment was converted into FXa concentrations from a standard
curve prepared with known dilutions of purified human FXa.
Thrombin generation on the surface of HUVECs
HUVECs were preincubated in 300 μl containing curcumin or
BDMC in 50 mM TrisHCl buffer, 100 pM FVa and 1 nM FXa
for 10 min and prothrombin was added to a final concen-
tration of 1 μM. After 10 min, duplicate samples (10 μl each)
were transferred to a 96-well plate containing 40 μl of 0.5 M
EDTA in Tris-buffered saline in each well to terminate pro-
thrombin activation. Activated prothrombin was determined
by measuring the rate of hydrolysis of S2238 measured at 405
nm. Dilutions with known amounts of purified thrombin were
used for standard curves.
Thrombin activity assay
Curcumin or BDMC in 50 mM TrisHCl buffer, pH 7.4 con-
taining 7.5 mM EDTA and 150 mM NaCl was mixed in the ab-
sence or presence with 150 μl of AT III (200 nM). After the
mixture was incubated at 37°C for 2 min, thrombin solution
(150 μl; 10 U/ml) was added and incubated at 37°C for 1 min.
Then, substrate for thrombin (S-2238, 150 μl; 1.5 mM) sol-
ution was added and absorbance at 405 nm was monitored for
120 s with a spectrophotometer (TECAN, Switzerland).
Factor Xa (FXa) activity assay
These assays were performed similar to the thrombin activity
assay. Instead of thrombin and S-2238, factor Xa (1 U ml/1)
and substrate S-2222 were used.
ELISA for PAI- 1
The concentrations of PAI-1 in HUVEC cultured supernatants
were determined by ELISA methods, according to the manu-
facturer’s recommended protocol (American Diagnostica Inc.,
Stamford, CT, USA).
Effect on bleeding time
The tail transection bleeding time was determined according
to the method of Dejana et al. (41) Male C57BL/6 mice were
fasted overnight and curcumin or BDMC was administered or-
ally to mice. One hour after administration, the mouse tail was
transected at 2 mm from the tip. Bleeding time was measured
as time elapsed until bleeding stopped. When bleeding time
lasted longer than 15 min, measurement was stopped and
bleeding time was recorded as 15 min for statistical analyses.
Statistical analysis
Data are expressed as the means ± standard deviation of at
least three independent experiments. Statistical significance
between two groups was determined by a Student’s t-test. The
significance level was set at P 0.05.
Acknowledgements
This work was supported by the National Research Foundation
of Korea (NRF) grant funded by the Korea government [MEST]
(No. 2011-003410, 2011-0026695, 2011-0030124).
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... 4. Curcumin possesses anti-thrombotic activities by inhibiting thrombin and FXa. It has been proven to be effective in reducing the viscosity of blood and can increase survival rates [2,9]. ...
... On the contrary, curcumin has an anti-inflammatory activity but it also possesses anti-thrombotic activities as it inhibits thrombin and FXa [9]. ...
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Curcumin, the bioactive ingredient of Curcuma longa (turmeric) has a wide range of therapeutic effects that make it an excellent candidate for use as adjuvant therapy in the treatment of patients with COVID or any viral pneumonia. Curcumin has potential antiviral effects, including protein binding affinity toward SARS-CoV-2 proteins. Preclinical studies have shown that curcumin effectively inhibits viral infection, alleviates the severity of lung injury by offsetting the cytokine storm, and inhibits subsequent fibrosis. Curcumin inhibits thrombin and FXa and reduces blood viscosity; it could therefore alleviate COVID or any post-viral thrombotic complications viz. pulmonary fibrosis, stroke, myocardial infarction, avascular necrosis of bone, and thereby increase survival benefits.
... CUR оказывает значительное влияние на гемостаз, выступая в качестве мощного антикоагулянта за счет ингибирования ключевых ферментов каскада свертывания крови -тромбина и фактора Ха (FXa) [124]. Антикоагулянтная активность CUR была подтверждена удлинением времени протромбинового времени (ПТ) и активированного частичного тромбопластинового времени (АЧТВ), а также ингибированием продукции FXa и последующего образования тромбина. ...
... Поскольку FXa, входя в состав протромбиназного комплекса, вызывает образование тромбина, который является конечным ферментом каскада свертывания крови и ответственен за образование сгустка и активацию тромбоцитов, ингибирование этих ферментов CUR обуславливает его потенциал как антикоагулянта [125,126]. Кроме того, CUR за счет ингибирования путей NF-κB и JNK в эндотелиальных клетках человека снижает продукцию TNF-α, что обуславливает уменьшение продукции PAI-1 (ингибитор активатора плазминогена-1), а это в свою очередь приводит к запуску активатора плазминогена и, следовательно, к усилению фибринолитической системы крови [124]. ...
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Curcumin, an active ingredient derived from turmeric root (Curcuma longa), has significant pharmacological properties including anti-inflammatory, antioxidant, antimicrobial, anticancer and analgesic activities. Studies show that curcumin affects the expression of various microRNAs and long non-coding RNAs to regulate cell proliferation and apoptosis in various types of malignant tumours. In addition, curcumin modulates signalling pathways such as PI3K/Akt/mTOR, MAPK/ERK and AMPK, activating autophagy and inhibiting tumour angiogenesis. It also inhibits metastasis and invasion of tumour cells by affecting epithelial-mesenchymal transition and expression of matrix metalloproteinases. Curcumin exhibits antibacterial and antiviral activity by disrupting bacterial cell membranes and inhibiting viral replication. The antioxidant properties of curcumin are due to its ability to neutralise reactive oxygen species and stimulate antioxidant enzymes. Curcumin also promotes wound healing by modulating inflammatory processes and stimulating angiogenesis. The analgesic effect of curcumin is due to its ability to stimulate the release of endogenous opioid peptides and modulate the activity of GABA receptors and ASIC and TRPV ion channels. Curcumin has an effect on lipid and carbohydrate metabolism, which makes it a promising agent for the treatment of dyslipidaemia and insulin resistance. The effect of curcumin on haemostasis is manifested in its ability to inhibit platelet aggregation and blood clotting, which may be useful for the prevention of cardiovascular diseases.
... Curcuminloaded solid lipid nanoparticles arrest the cell cycle at G1/S and reduce the expression of cyclin D1 and Cyclin-Dependent Kinase 4 (CDK4), leading to the strong induction of apoptosis and ROS production in vitro; specifically, they induce apoptosis by activating P53 and P21 proteins, which regulate the Phosphoinositide 3-Kinase-Protein Kinase B (PI3K-AKT) and Nuclear Factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling pathways [33,55]. However, as no studies on humans are available and since curcumin may exert an anticoagulant effect by inhibiting the activities of Factor Xa (FXa) and thrombin, its supplementation may be hazardous in cancer patients [56]. ...
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Among the distressing side effects of cancer treatments, hair loss is one of the most disturbing for the quality of life and adherence to therapy in breast cancer patients. Many patients take nutritional supplements to prevent hair loss or enhance regrowth. Based on their mechanism and timing of use, nutritional supplements could be divided into safe, cautious, debated, and contraindicated categories. Non-contraindicated supplements generally include safe supplements like vitamin D, which is not known to interfere with cancer treatments. Those that are contraindicated include phytoestrogens and compounds affecting estrogen pathways because of the risk of stimulating tumor growth in cancers sensitive to estrogen. Antioxidants like tocotrienols and resveratrol are given judiciously because of potential interference with cancer therapies dependent on reactive oxygen species. Supplements debated, including nicotinamide, folate, and iron, pose a risk by promoting cellular proliferation or altering the tumor microenvironment. Biotin is nontoxic but interferes with blood test results and is thus difficult in cancer monitoring. Evidence regarding nutritional supplements’ safety and efficacy in this context is conflicting. Management by an oncologist is required along with more studies to clearly establish the safety parameters and efficacy guidelines.
... Despite their beneficial properties, plant compounds can interact with various medications, including anticoagulants, antihypertensive drugs, and insulin-sensitizing agents [38]. For example, compounds like curcumin and resveratrol may have blood-thinning effects, which can enhance the effects of anticoagulants, increasing the risk of bleeding [39]. Similarly, some plant compounds may interact with medications that regulate blood pressure or glucose levels, potentially leading to additive or synergistic effects [40]. ...
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Metabolic syndrome (MetS) is a group of interrelated risk factors, including central obesity, insulin resistance, dyslipidemia, and hypertension that increase the risk of developing cardiovascular diseases, type 2 diabetes, and other chronic conditions. The global prevalence of MetS has been steadily rising, largely driven by sedentary lifestyles and poor dietary habits. While conventional pharmacological treatments are essential for managing MetS, there is growing interest in plant-based compounds as potential therapeutic agents. These compounds offer promising benefits due to their ability to address multiple aspects of MetS, including inflammation, oxidative stress, insulin resistance, and lipid metabolism, with minimal side effects. This review provides an overview of several plant-based compounds, such as curcumin, resveratrol, berberine, green tea catechins, and cinnamon, highlighting their mechanisms of action, clinical evidence supporting their use, and potential for future treatment options. By exploring the efficacy of these natural compounds, this review aims to contribute to the understanding of alternative and complementary approaches in managing MetS, with a focus on their therapeutic potential, safety, and clinical applicability.
... Благодаря наличию такого механизма действия куркумин может усиливать антиагрегантное действие Аспирина и других антиагрегантов. В дополнение к этому куркумин способен подавлять активность тромбина и фактора свертывания Ха, что может усилить действие антикоагулянтов (Гепарин и др.) [70][71][72][73][74]. ...
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This review presents current data on the pharmacokinetics, side effects, dosage forms and interactions of curcumin. Curcumin, the major bioactive component of turmeric, has low bioavailability due to its low water solubility, rapid metabolism in the liver and rapid excretion from the body. The main pathways of curcumin metabolism are described, including its reduction and subsequent conjugation with glucuronic acid and sulphates. Side effects of curcumin such as hyperoxaluria, iron deficiency anaemia, hepatotoxicity, arrhythmias, allergic reactions and potential carcinogenic properties are discussed. Various dosage forms of curcumin developed to enhance its bioavailability are discussed, including liposomes, nanoparticles, hydrogels and phytosomes. Particular attention is given to the drug interactions of curcumin with chemotherapeutic agents such as 5-fluorouracil, vincristine, gemcitabine, adriamycin and cisplatin, as well as with hypolipidaemic agents, antiaggregants and anticoagulants. These data highlight the need for further studies to optimise the therapeutic use of curcumin and minimise its side effects.
... Curcumin is a phytopolyphenol obtained from the Curcuma longa plant [40]. This bioactive compound is a multifunctional active agent known with antibacterial [41], antifungal [42], antiviral [43], antioxidant [44], anti-inflammatory [44], anticoagulant [45], antiatherosclerotic [46], anticarcinogenic [47], and hypoglycemic [48] effects. Due to these therapeutic effects, it is preferred for use in pharmaceutical and biomedical fields. ...
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Sutures provide mechanical support for wound closure after various traumas and surgical operations. Absorbable sutures are increasingly favored as they eliminate the need for secondary procedures and minimize additional damage to the wound site. In this study, chitosan sutures were produced using the dry jet–wet spinning method, achieving number 7-0 sutures (approximately 76 μm diameter) with a homogeneous surface. FTIR analysis demonstrated molecular interactions between chitosan and TiO2 or curcumin, confirming successful incorporation. The addition of 3% TiO2 increased the tensile strength of chitosan sutures by 12.32%, reaching 189.41 MPa. Morphological analysis revealed smooth surfaces free of pores and bubbles, confirming the production of high-quality sutures. Radical scavenging activity analysis showed that curcumin-loaded sutures exhibited 43% scavenging ability after 125 h, which was significantly higher compared to pure chitosan sutures. In vitro antibacterial tests demonstrated that curcumin-loaded sutures provided 98.87% bacterial inactivation against S. aureus within 24 h. Additionally, curcumin release analysis showed a cumulative release of 77% over 25 h. The bioactivity of the sutures was verified by hydroxyapatite layer formation after incubation in simulated body fluid, supporting their potential for tissue regeneration. These findings demonstrate that TiO2 reinforcement and curcumin loading significantly enhance the functional properties of chitosan sutures, making them strong candidates for biocompatible and absorbable surgical applications.
... Maizure [31] reported that ginger is richer in polyphenols than turmeric. Also, the study by Kim et al. [32] on a set of spices, showed that coriander is the richest in polyphenols, followed by caraway, turmeric, fennel and cumin. ...
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Curcuma longa (turmeric) has a long history of use in Ayurvedic medicine as a treatment for inflammatory conditions. Turmeric constituents include the three curcuminoids: curcumin (diferuloylmethane; the primary constituent and the one responsible for its vibrant yellow color), demethoxycurcumin, and bisdemethoxycurcumin, as well as volatile oils (tumerone, atlantone, and zingiberone), sugars, proteins, and resins. While numerous pharmacological activities, including antioxidant and antimicrobial properties, have been attributed to curcumin, this article focuses on curcumin's anti-inflammatory properties and its use for inflammatory conditions. Curcumin's effect on cancer (from an anti-inflammatory perspective) will also be discussed; however, an exhaustive review of its many anticancer mechanisms is outside the scope of this article. Research has shown curcumin to be a highly pleiotropic molecule capable of interacting with numerous molecular targets involved in inflammation. Based on early cell culture and animal research, clinical trials indicate curcumin may have potential as a therapeutic agent in diseases such as inflammatory bowel disease, pancreatitis, arthritis, and chronic anterior uveitis, as well as certain types of cancer. Because of curcumin's rapid plasma clearance and conjugation, its therapeutic usefulness has been somewhat limited, leading researchers to investigate the benefits of complexing curcumin with other substances to increase systemic bioavailability. Numerous in-progress clinical trials should provide an even deeper understanding of the mechanisms and therapeutic potential of curcumin.
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In blood coagulation on endothelium, an unperturbed vascular endothelial cell surface apparently provides activity equivalent to the phospholipid needed for generation of factor Xa and thrombin in soluble systems. Phospholipid in soluble systems also markedly enhances the ability of factor Xa to activate factor VII; therefore we investigated the influence of an unperturbed monolayer of human umbilical vein endothelial cells (HUVEC) upon factor VII activation. HUVEC were found to augment factor Xa-catalyzed activation of factor VII. This appeared to result from the binding of trace amounts of factor Xa to the cells. Adding active site-inhibited factor Xa to reaction mixtures, but not factor X, abolished the enhanced activation. Adding either anti-factor V antibodies or exogenous factor Va had no effect upon reaction rates. Thus factor Va does not function as a cofactor for the reaction. In further experiments the effect upon activation of factor VII and prothrombin was studied by varying the order of addition of factor Xa and factor Va to supernatants of HUVEC monolayers. Evidence was obtained that HUVEC, unlike platelets, possess a functional factor Xa binding site that is independent of factor Va. Since phospholipid is the only known cofactor for factor Xa/Ca2+-induced activation of factor VII, the demonstration of enhanced activation of factor VII in the presence of unperturbed cultured HUVEC supports a hypothesis that the functional equivalent of procoagulant phospholipid is available on the luminal surface of vascular endothelium in vivo.
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Atherosclerosis preferentially occurs in areas of turbulent flow and low fluid shear stress, whereas laminar flow and high shear stress are atheroprotective. Inflammatory cytokines, such as tumor necrosis factor-α (TNF), have been shown to stimulate expression of endothelial cell (EC) genes that may promote atherosclerosis. Recent data suggest that steady laminar flow decreases EC apoptosis and blocks TNF-mediated EC activation. EC apoptosis is likely important in the process termed “plaque erosion” that leads to platelet aggregation. Steady laminar flow inhibits EC apoptosis by preventing cell cycle entry, by increasing antioxidant mechanisms (e.g., superoxide dismutase), and by stimulating nitric oxide-dependent protective pathways that involve enzymes PI3-kinase and Akt. Conversely, our laboratory has identified nitric oxide-independent mechanisms that limit TNF signal transduction. TNF regulates gene expression in EC, in part, by stimulating mitogen-activated protein kinases (MAPK) which phosphorylate transcription factors. We hypothesized that fluid shear stress modulates TNF effects on EC by inhibiting TNF-mediated activation of MAP kinases. To test this hypothesis, we determined the effects of steady laminar flow (shear stress = 12 dynes/cm2) on TNF-stimulated activity of two MAP kinases: extracellular signal regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK). Flow alone stimulated ERK1/2 activity, but decreased JNK activity compared to static controls. TNF (10 ng/ml) alone activated both ERK1/2 and JNK maximally at 15 minutes in human umbilical vein EC (HUVEC). Pre-exposing HUVEC for 10 minutes to flow inhibited TNF activation of JNK by 46%, but it had no significant effect on ERK1/2 activation. Incubation of EC with PD98059, a specific mitogen-activated protein kinase kinase inhibitor, blocked the flow-mediated inhibition of TNF activation of JNK. Flow-mediated inhibition of JNK was unaffected by 0.1 mM L-nitroarginine, 100 μM 8-bromo-cyclic GMP, or 100 μM 8-bromo-cyclic AMP. Transfection studies with dominant negative constructs of the protein kinase MEK1 and MEK5 suggested an important role for BMK1 in flow-mediated regulation of TNF signals. In summary, the atheroprotective effects of steady laminar flow on the endothelium involve multiple synergistic mechanisms.
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Numerous acquired hemostatic abnormalities have been identified in renal insufficiency. Hemodialysis procedures add to these disturbances as they repetitively imply turbulent blood flow, high shear stress, and contact of blood to artificial surfaces. This nonphysiological environment leads to activation of platelets, leukocytes, and the coagulation cascade, resulting in fouling of the membrane and ultimately in clotting of fibers and the whole hemodialyzer. Anticoagulation in hemodialysis is targeted to prevent this activation of coagulation during the procedure. Most agents inhibit the plasmatic coagulation cascade. Still commonly used is unfractionated heparin, followed by low-molecular-weight heparin preparations with distinct advantages. Immune-mediated heparin-induced thrombocytopenia constitutes a potentially life-threatening complication of heparin therapy requiring immediate switch to nonheparin alternative anticoagulants. Danaparoid, lepirudin, and argatroban are currently being used for alternative anticoagulation, all of which possess both advantages and limitations. In the past, empirical strategies reducing or avoiding heparin were applied for patients at bleeding risk, whereas nowadays regional citrate anticoagulation is increasingly used to prevent bleeding by allowing procedures without any systemic anticoagulation. Avoidance of clotting within the whole hemodialyzer circuit is not granted. Specific knowledge of the mechanisms of coagulation, the targets of the anticoagulants in use, and their respective characteristics constitutes the basis for individualized anticoagulation aimed at achieving full patency of the circuit throughout the procedure. Patency of the circuit is an important prerequisite for optimal hemodialysis quality.
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The tumor suppressor gene Wnt inhibitory factor-1 (WIF-1) has been found to be promoter hypermethylated and silenced in lung cancer cell lines and tissues. Curcuminoids are major active components of the spice turmeric, and have recently been reported to be potential hypomethylation agents. In the present study, the hypomethylation effects of three major curcuminoids, curcumin, demethoxycurcumin and bisdemethoxycurcumin, were compared in vitro using ELISA, and their demethylation potential was confirmed by methylation-specific PCR. It was found that bisdemethoxycurcumin possesses the strongest demethylation function in vitro compared to the other two curcuminoids, exerting its effect at a minimal demethylation concentration of 0.5-1 µM. The WIF-1 promoter region was demethylated after treatment with 20 µM demethoxycurcumin and bisdemethoxycurcumin, but failed to respond to 20 µM curcumin. In the A549 cell line, RT-PCR and Western blotting were used to confirm that WIF-1 expression was restored after curcuminoid-induced promoter hypermethylation. Since the results regarding the demethylation potential of the three major curcuminoids to restore WIF-1 expression indicated that bisdemethoxycurcumin has the strongest hypomethylation effect, this curcuminoid may have therapeutic use in the restoration of WIF-1 expression in NSCLC.
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Curcuminoids are the major active components extracted from Curcuma longa and are well known for their antioxidant effects. Previous studies have reported that the antioxidant properties of curcuminoids are mainly attributed to their free radical scavenging abilities. However, whether there are other mechanisms besides the non-enzymatic process and how they are involved, still remains unknown. In the present study, we explored the protective effects of bisdemethoxycurcumin (Cur3) against tert-butyl hydroperoxide (t-BHP)-induced cytotoxicity in human umbilical vein endothelial cells (HUVECs), focusing on the effect of Cur3 on the regulation of the phosphatidylinositol 3-kinase (PI3K)/Akt and the mitogen-activated protein kinase (MAPK) pathways. The pre-treatment with Cur3 inhibited t-BHP-induced cell damage dose-dependently, which was evident by the increased cell viability and the corresponding decrease in lactate dehydrogenase release. The pre-treatment with Cur3 also attenuated t-BHP-induced cell morphological changes and apoptosis. MAPKs, including p38, c-Jun N-terminal kinase (JNK), extracellular signal-regulated protein kinase 1/2 (ERK1/2), as well as PI3K/Akt have been reported to be involved in proliferation, apoptosis and differentiation under various stress stimulations. The pre-treatment with Cur3 decreased t-BHP-induced ERK1/2 phosphorylation and increased t-BHP-induced Akt phosporylation but did not affect the phosphorylation of p38 or JNK. In addition, the Cur3-induced increase in cell viability was attenuated by the treatment with wortmannin or LY294002, the upstream inhibitors of Akt, and was enhanced by the treatment with 2-[2'-amino-3'-methoxyphenyl]-oxanaphthalen-4-one (PD98059), an upstream inhibitor of ERK1/2. These results suggest that the ERK1/2 and PI3K/Akt signaling pathways could be involved in the protective effects of Cur3 against t-BHP-induced damage in HUVECs.
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Curcumin is the main bioactive constituent derived from the rhizome of turmeric (Curcuma longa Linn.), which has been used traditionally as hepatoprotective agents in ayurvedic and traditional Chinese medicine for centuries. The present study was carried out to demonstrate the potential protective effect of curcumin pretreatment against ethanol-induced hepatocytes oxidative damage, with emphasis on heme oxygenase-1 (HO-1) induction. Rat primary hepatocytes were isolated and treated with ethanol (100mM) and diverse doses of curcumin (0-50 microM), which was pretreated at various time points (0-5h) before ethanol administration. Hepatic enzyme releases in the culture medium and redox status including HO-1 enzyme activity were detected. Ethanol exposure resulted in a sustained malondialdehyde (MDA) elevation, glutathione (GSH) depletion and evident release of cellular lactate dehydrogenase (LDH) and aspartate aminotransferase (AST), which was significantly ameliorated by curcumin pretreatment. In addition, dose- and time-dependent induction of HO-1 was involved in such hepatoprotective effects by curcumin. Curcumin exerts hepatoprotective properties against ethanol involving HO-1 induction, which provide new insights into the pharmacological targets of curcumin in the prevention of alcoholic liver disease.