ArticlePDF Available

Pro-coagulant Activity of Phenolic Acids Isolated from Blumea Riparia

SAGE Publications Inc
Natural Product Communications
Authors:

Abstract

The effects of extracts of the aerial part of Blumea riparia DC. and their phenolic acids on hemostasis were evaluated. The EtOAc fraction showed significantly reduced blood clotting time (CT) and tail bleeding time of transection (BT) of mice in vivo. This fraction contained vanillic acid (1), syringic acid (2), p-coumaric acid (3), caffeic acid (4), and protocatechuic acid (5). Compound 1 reduced prothrombin time (PT), and strengthened mice uterine contractions. Compound 3 reduced CT and the activated partial thromboplastin time (APTT). Compound 5 reduced CT and increased the frequency of mice uterine contraction in a dose-dependent manner. Compound 2 reduced APTT. Compound 4 remarkably strengthened uterine contraction. Taken together, these data suggest that compounds 1, 3, and 5 possess procoagulant activity which jointly synergize blood coagulation via different mechanisms.
Pro-coagulant Activity of Phenolic Acids Isolated from
Blumea riparia
Li Huanga,c, Cuiwu Lina ,*, Aiyuan Lib, Baoyao Weic, Jianwen Tengc and Lue Lia
aDepartment of Chemistry and Chemical Engineering, Guangxi University, Nanning,
Daxue Road, 88, 530004, People’s Republic of China
bGuang Xi Traditional Chinese Medical University, Nanning, Mingxiu East Road, 88, 530021,
People’s Republic of China
cInstitute of Light Industry and Food Engineering, Guangxi University, Daxue Road, 88,
530004, People’s Republic of China
lincuiwu@126.com
Received: April 15th, 2010; Accepted: June 9th, 2010
The effects of extracts of the aerial part of Blumea riparia DC. and their phenolic acids on hemostasis were evaluated. The
EtOAc fraction showed significantly reduced blood clotting time (CT) and tail bleeding time of transection (BT) of mice in
vivo. This fraction contained vanillic acid (1), syringic acid (2), p-coumaric acid (3), caffeic acid (4), and protocatechuic acid
(5). Compound 1 reduced prothrombin time (PT), and strengthened mice uterine contractions. Compound 3 reduced CT and the
activated partial thromboplastin time (APTT). Compound 5 reduced CT and increased the frequency of mice uterine
contraction in a dose-dependent manner. Compound 2 reduced APTT. Compound 4 remarkably strengthened uterine
contraction. Taken together, these data suggest that compounds 1, 3, and 5 possess procoagulant activity which jointly
synergize blood coagulation via different mechanisms.
Keywords: Blumea riparia, procoagulant, blood, hemostasis, phenolic acids, coagulant.
Menorrhagia and postpartum hemorrhage are common
illnesses that affect many women [1a, 1b]. Oxytocin,
ergot alkaloids, prostaglandins, tranexamic acid,
ethamsylate, and recombinant activated factor VII are
used for treatment but produce adverse reactions [1c].
Some compounds isolated from plants display
hemostatic activity. For example, an extract from
Blumea riparia (BL.) DC. (Compositae) is used in
traditional Chinese medicine for reducing uterus
blood, preventing gynecological inflammation, and
accelerating body recovery postpartum. In addition, an
herbal medicine produced from B. riparia DC. is
clinically used for the treatment of postabortal
metrorrhagia, and other gynecological problems [2a,2b].
Previous chemical studies of this plant characterized
flavones, organic acids, acetylenes, and xanthene
sesquiterpenes [3a-3d], while the present study, found
several additional constituents and evaluated their
procoagulant activity and structure-activity relationship.
Our findings provide a molecular basis for the clinical
application of B.riparia in the treatment of menorrhagia
and postpartum hemorrhage.
To analyze the procoagulant activity of B. riparia, the
effects of orally administered fractions Fa, Fb, and Fc
on blood clotting time (CT) and tail transection bleeding
time (BT) in rats were examined. As shown in Table 1,
a dose of 1.08 and 2.16 g /kg Fb significantly reduced
CT and BT (P<0.05), in comparison with the control
group. Fa, Fb, and Fc exhibited a biphasic effect on the
blood coagulation, by which the CT and BT increased at
a dose of 4.32 g /kg. Similarly, polysaccharides from
green seaweeds also showed a dual hemostatic effect,
that is, inhibition or activation of coagulation pathways
at different doses [4a]. These results give a scientific
explanation for the folkloric use of B. riparia as a
hemostatic in gynecology, as well as an inhibitor of
thrombus formation at larger doses.
As Fb exhibited better hemostatic activity than the other
fractions, it was fractionated to give the five phenolic
acids, vanillic (1), syringic (2), p-coumaric (3), caffeic
(4), and protocatechuic (5). The hemostatic activities of
these compounds were examined by CT, prothrombin
time (PT) and activated partial thromboplastin time
(APTT), and uterine contractility, which are commonly
used for evaluating hemostatic agents. As large amounts
NPC Natural Product Communications 2010
Vol. 5
No. 8
1263 - 1266
1264 Natural Product Communications Vol. 5 (8) 2010 Huang et al.
Table 1: Effect of the different fractions on procoagulant activity in mice
in vivo.
Groups Dose (g/kg) CT(min) BT(min)
control - 7.83±2.00 3.89±1.32
Ethamsylate 0.25 3.73±0.94** 1.28±0.51**
4.32 6.19±2.25 2.72±1.48
2.16 5.45±2.01 2.34±0.94
Fa
1.08 5.50±1.36 1.98±1.13
4.32 6.36±1.93 3.40±1.89
2.16 4.57±1.35** 1.65±0.68*
Fb
1.08 4.78±1.56* 1.73±1.48*
4.32 8.52±1.54 2.64±0.78
2.16 9.06±1.05 2.52±1.18
Fc
1.08 5.58±1.56 3.00±1.62
Negative control, saline solution; positive control, ethamsylate. Values
are mean±SD. (n = 8).Statistical comparison was performed using
ANOVA followed by Tukey’s test. * Significant vs. the control group (p
<0.05); ** Significant vs. the control group (p <0.01)
of test samples were required for the week- long daily
oral administration, sufficient quantities of compounds
15 had to be purchased. Interestingly, at a dose of
3 mmol/kg, only compound 3 (Figure 1) significantly
decreased CT (P <0.05), suggesting that, while
compound 3 may be effective, other procoagulant
components may be present to account for the high dose
of compound 3 needed.
Blood coagulation results from a series of proteolytic
reactions involving the step-wise activation of
coagulation factors I-XII. Subsets of these factors can
be activated by two distinct pathways, the extrinsic and
the intrinsic pathway.[4b] The APTT is commonly used
for determining the overall efficiency of the intrinsic
coagulation pathway, while PT is the screening test for
the extrinsic coagulation pathway [4c]. The APTT was
reduced to12.5, 13.3, and 15.6% in mice treated with
compounds 2, 3, and 5 when compared to the control
group. However, the PT values remained unchanged in
the presence or absence of these compounds (Table 2).
These data suggest that the intrinsic coagulation
pathway may be the target of these compounds.
Protocatechuic acid (5) can enhance the activity of
anticoagulant factors such as antithrombin-III and
protein C [4d].. This observation may be explained if it
has dual functions in the coagulation system. It may act
as a procoagulant to promote thrombus when bleeding
occurs and also act as an anticoagulant when combined
with the cofactor thrombomodulin in the protein case
complex [5a, 5b].
Vanillic acid (1) significantly lowered PT, but had no
effect on APTT. The lessening of PT suggested that 1
could activate the extrinsic coagulation pathway. A
similar observation showed that 1 had weak
antithrombotic effects when evaluated for thrombosis in
the mice model [6a]. It also decreased the recalcification
time in a dose-dependent manner and inhibited
5’nucleotidase activity specifically, which mediated the
Figure 1: Effect of phenolic acids on CT in mice in vivo.
Negative control, saline solution; positive control, ethamsylate. The
treatment with compound 15 were used in a dose of 0.2mmol/kg or
3mmol/kg, respectively. Values are mean±SD. (n = 8). Statistical
comparison was performed using ANOVA followed by Tukey’s test.
* Significant vs. the control group (p <0.05).
Table 2: Effect of phenolic acids on PT and APTT in vivo.
Groups Dose( mmol/kg) PT (s) APTT(s)
control - 9.61±1.72 19.69±1.49
Positive control 1(g·kg-1) 7.23±0.80** 15.38±0.81**
1 0.75 7.38±1.29* 18.31±2.37
2 0.75 7.77±1.62 17.22±1.53*
3 0.75 7.88±0.80 17.08±1.93*
4 0.75 8.16±1.30 18.28±2.36
5 0.75 7.94±0.75 16.60±1.43**
Negative control, saline solution; positive control, Yunnan white power.
PT values are mean±SD. (n = 8) and APTT values are mean±SD. (n =
10). Statistical comparison was performed using ANOVA followed by
Tukey’s test. * Significant vs. the control group (p <0.05); ** Significant
vs. the control group (p <0.01).
anticoagulant effect of Naja naja venom [6b] Therefore,
B. riparia DC. may affect both intrinsic and extrinsic
pathways to regulate heavy menstrual bleeding or
postpartum hemorrhage.
The effects of different phenolic acids on spontaneous
contractile activity of uterine smooth muscle in estrous
mice were compared. As summarized in Table 3, there
was no obvious difference in the tension of uterine
contraction before and after the addition of phenolic
acids to the bath, except for compounds 2 and 4.
Surprisingly, compounds 4 and 5 significantly increased
the frequency of uterine contraction in a dose-dependent
manner (P < 0.01). Compound 1 also accelerated the
frequency of uterine contraction at 1.43×10-4 mmol/ml
(P < 0.05). These results indicate that phenolic acids can
strengthen the frequency of mice uterine contractions,
while only affecting tension slightly. Alteration of
uterine contractions by drugs is important in obstetrics
practice. Inadequate uterine contractions could impede
labor and increase the incidence of emergency caesarian
sections, whereas promotion of uterine contractions
could prevent or treat excessive hemorrhage in the
immediate postpartum period [7]. Based on the above it
may be assumed that 4 and 5 strengthen contraction
of uterine and blood vessel to promote the repair and
Pro-coagulant activity of phenolic acids Natural Product Communications Vol. 5 (8) 2010 1265
Table 3: Effect of phenolic acids on the uterine contraction of mice in vitro.
Frequency of uterine
contractioncycle/5min Tension of uterine contraction
n ( g
groups
dose×10-
4mmol/
mL Before After Before After
control - 6.26±2.14 6.21±2.20 1.80±0.54 1.77±0.49
positive - 5.25±1.39 15.25±3.81** 1.45±0.46 3.43±0.92**
0.71 11.88±5.03 14.75±5.12 1.96±0.73 2.03±0.75
1.43 10.13±3.27 12.25±4.27* 1.59±0.58 1.55±0.55 1
2.14 11.13±4.49 11.75±3.24 1.86±0.70 1.70±0.48
0.36 12.63±4.87 13.75±5.06 2.15±0.72 2.24±0.74
0.71 10.38±3.20 11.00±4.90 1.72±0.67 1.93±0.66* 2
1.43 8.75±4.92 9.50±4.41 1.65±0.55 2.19±0.74
0.71 9.50±2.45 9.75±4.27 1.85±0.64 1.87±0.69
1.07 8.75±4.37 8.50±4.21 1.37±0.10 1.70±0.49
3
1.43 7.88±2.47 8.50±4.69 1.70±0.49 1.96±0.66
0.71 7.88±3.68 13.00±5.95* 3.08±1.06 3.16±1.09
1.43 9.13±4.16 18.50±6.19* 2.06±0.72 2.80±1.00
4
2.86 5.25±1.67 23.13±10.62** 1.73±0.48 2.61±0.83*
0.89 10.13±3.48 14.25±4.65* 2.30±0.59 2.25±0.69
1.07 6.75±2.60 15.38±3.78** 2.11±0.59 2.41±0.56
5
1.25 7.13±2.80 16.13±6.96** 2.45±0.76 2.56±0.84
Negative control, saline solution; positive control, oxytocin and dose is 1×10-
3unit/mL.Values are mean±S.D. (n = 8). The uterine contractions were
recorded for 10 min immediately before and after addition of phenolic acids to
the bath. *Contraction after addition of phenolic acids is significantly different
from the values before its addition to the organ bath at p <0.05 using paired t-
test with a two-tailed p-value. **Contraction after addition of phenolic acids is
significantly different from the values before its addition to the organ bath at p
<0.01 using paired sample t-test with a two-tailed p-value.
regeneration of tissue, thus preventing hemorrhage. B.
riparia DC. can therefore not only be used for
postpartum care, but also for treating irregular, painful,
or excessive menstruation, uterine pain and dysfunction.
Experimental
General: The ethanol extract of B. riparia DC. was
provided by Guixi Pharmacy Co., Ltd. of Naning,
Guangxi, China. Caffeic acid (4) was purchased from
Shanghai Junchuang Biological Tech. co., (Shanghai,
China). p-Coumaric acid (3), vanillic acid (1), syringic
acid (2) and protocatechuic acid (5) were purchased
from Wuhan Hezhong Bio-Chemical Manufacture Co.,
(Wuhan China). APTT assay kit and PT assay kit were
purchased from Beijing Bo Lai Bio. Co. (Beijing,
China). Column chromatography was performed on
Silica gel (200–300 mesh), gel H (Qingdao Mar. Chem.
Ind. Co.Ltd), and Sephadex LH-20 (Pharmacia).
Extraction and sample preparations: The ethanol
extract of (4.5 kg) B. riparia. was evaporated under
reduced pressure to obtain a residue (3.2 kg), which was
diluted with water (6.4 kg) and extracted several times
with petroleum ether. The petroleum ether extracts were
combined to form fraction (Fa). The aqueous layer was
further extracted with EtOAc to give fraction (Fb) and
the residual aqueous layer gave fraction (Fc). All these
fractions were evaporated under reduced pressure to
give dried products, Fa (142.7 g), Fb (286.7 g), and
Fc (2520 g).
Among the extracts and fractions tested for their
procoagulant activities (Table 1), Fb showed significant
inhibition activities. Fb (100 g) was repeatedly
chromatographed over a silica-gel to give vanillic acid
(1) (14 mg) and syringic acid (2) (18 mg), p-coumaric
acid (3) (22 mg), caffeic acid (4) (54 mg) and
protocatechuic acid (5) (36 mg). These phenolic acids
were identified by comparison of physical and
spectroscopic data with literature data. [8a-8d]
Animals: Mice (Kingming strain) weighing 18–22 g
were used to assay CT and BT in vivo. Female mice
(Kingming strain) weighing 25–30 g were used to assay
uterine contractility. New Zealand rabbits weighing
1.8–2.2 kg were used to assay PT and APTT in vivo.
The animal experiments were conducted in accordance
with international guidelines.
Assay of blood clotting time in vivo (CT): The blood
clotting time was evaluated by the glass slide method
[9a]. The fractions (Fa, Fb, and Fc) and phenolic acids
were dissolved in water. Experimental mice were
divided into 25 groups of 8 and administrated with the
fractions or reagents at 0.1ml/10g by an oral gavage
with a syringe. The control groups were treated with
saline solution. The positive groups were injected with
ethamsylate (Beijing Pharmaceutical Factory, Beijing,
China) at 0.25g/kg. After three days, each mouse’s
inner canthus was punctured with a glass capillary and a
drop of blood from the supraorbital vein was collected
on a glass slide. The bleeding time was recorded
between blood collection and blood streak formation.
Tail transection bleeding time in vivo (BT):
Experimental mice were divided into 11 groups each
consisting of 8 mice. The preparations of the fractions,
control group, positive groups and the manner of
administration are similar to that described above for
the CT assay. After three days, the mice were
anesthetized with sodium pentobarbital. The bleeding
time was recorded according to the method by Dejana
et al. [9b].
Assay for prothrombin time (PT) and activated
partial thromboplastin time (APTT) in vivo:
Experimental rabbits were divided into 8 groups of 3
and 10 ml/kg of the reagent administered with a syringe
by an oral gavage for once a day for three days at the
same time. The positive groups were given 1g/kg
Yunnan white power (Yunnan Baiyao Group Co., Ltd.,
Yunnan, China). Rabbit blood was collected with
plastic disposable syringes 1h after the rabbits were
administered in the third day. Fresh blood was mixed
with 1:9 volume of 3.2% sodium citrate. Platelet poor
plasma was obtained by centrifuging citrated blood for
15 min at 3000 × g. PT and APTT were determined
immediately using a kit according to the cited method
1266 Natural Product Communications Vol. 5 (8) 2010 Huang et al.
[9c] All experiments were carried out in at least
duplicates.
Measurement of uterine contractility in vitro: Female
mice were injected subcutaneously with 0.1mg/kg
estradiol (Mingxin Pharmaceutical Co., Ltd.,
Guangzhou, China). After two days, the mice were
sacrificed by decapitation. Both uterine horns were
isolated, cut in half, and placed in isolated organ baths
incubated with De Jalon’s solution bubbled with
carbogen mixture (95% O2 and 5% CO2) at 37±0.5ºC.
Each uterine horn was subjected to a resting tension of
0.5-1g and equilibrated for at least 30-45 min before the
reagents were added to the organ baths. The positive
groups were treated with oxytocin 1×10-3 unit/mL. The
mice uterus contractions were recorded by polygraph
system (HW-400S) (Chengdu, China.) [9d].
Statistical analysis: Data are expressed as
mean±SD(standard deviations), and statistical
significance was determined by one-way analysis of
variance (ANOVA) and Tukey’s test. In the
measurement of uterine contractility, the differences
between before and after addition of samples were
tested for paired t-test with a two-tailed p-value. A
value of p<0.05 was considered significant.
Acknowledgments - We are grateful to Guixi
Pharmacy Co., Ltd., Guangxi, China for providing raw
materials for experimentation. This work was supported
by a major grant from the National Natural Science
Foundation of China (20962002 and 20662001), a
major grant from the Key Science Project of Guangxi
Province (04080005), and an innovative pilot scheme
for the College Students of China (200714).
References
[1] (a) Carroli G, Cuesta C, Abalos E, Gulmezoglu A. (2008) Epidemiology of postpartum haemorrhage: a systematic review. Best
Practice & Research Clinical Obstetrics & Gynaecology, 22, 999–1012; (b) Burns S, Parapia LA. (2005) Haematological causes of
menorrhagia. Reviews in Gynaecological Practice, 5, 8–14; (c) Bolte AC, Bouma L, Geijn HPv. (2005) Medical therapies for
primary postpartum hemorrhage. International Congress Series, 1279, 364–368.
[2] (a) Zhang X. (2002) A Clinical Observation on Fuxue Kang Granule in the treatment of postabortal metrorrhagia. Traditional
Chinese Drug Research & Clinical Pharmacology, 13, 6–8, 64; (b) Zhou Q, Bian DH, Li FL, Zhu JH. (1999) The observation of
the effect of Fu Xue Kang granule on vaginal bleeding after drug-induced abortion. Acta Universitatis scientiae medicinae
Chongqing, 24, 423–424.
[3] (a) Cao JQ, Dang Q, Fu HW, Yao Y, Pei YH. (2007) Isolation and identification of chemical constituents from Blumea riparia DC. .
Journal of Shenyang Pharmaceutical University, 24, 615–618; (b) Cao JQ, Sun SW, Cheng H, Wang YN, Pei YH. (2008) Studies
on flavonoids from Blumea riparia. China Journal of Chinese Materia Medica, 33, 782–784; (c) Huang L, Lei T, Lin CW, Kuang
XC, Chen HY, Zhou H. (2010) Fitoterapia, 81, 389–392; (d) Cao JQ, Yao Y, Chen H, Qiao L, Zhou YZ, Pei YH. (2007) A new
xanthene from Blumea riparia. Chinese Chemical Letters, 18, 303–305.
[4] (a) Ciancia M, Quintana I, Vizcargüénaga MI, Kasulin L, Dios AD, Estevez JM, Cerezo AS. (2007) Polysaccharides from the green
seaweeds Codium fragile and C. vermilara with controversial effects on hemostasis. International Journal of Biological
Macromolecules, 41, 641–649; (b) Butenas S, Mann KG. (2002) Blood coagulation. Biochemistry (Moscow), 67, 3–12; (c) Carville
DGM, Guyer KE. (1998) Coagulation testing (part 1): current methods and challenges. IVD Technology, 59; (d) Lin C, Huang C,
Huang C, Yin M. (2009) Anticoagulatory, antiinflammatory, and antioxidative effects of protocatechuic acid in diabetic mice.
Journal of Agricultural and Food Chemistry, 57, 6661–6667.
[5] (a) Cartwell M, Cera ED. (2000) Rational design of a potent anticoagulant thrombin. Journal of Biological Chemistry, 275,
39827–39830; (b) Chan KC, Yin MC, Chao WJ. (2007) Effect of diallyl trisulfide-rich garlic oil on blood coagulation and plasma
activity of anticoagulation factors in rats. Food Chemistry and Toxicology, 45, 502–507.
[6] (a) Yun-Chol HS, Kim MH, Jung KH. (1996) Esters of substituted benzoic acids as anti-thrombotic agents. Archives of Pharmacal
Research, 19, 66–70; (b) Dhananjaya BL, Nataraju A, Rajesh R, Raghavendra Gowda CD, Sharath BK, Vishwanath BS, D'Souza
CJM. (2006) Anticoagulant effect of Naja naja venom 5'-nucleotidase: demonstration through the use of novel specific inhibitor,
vanillic acid. Tox i con, 48, 411–421.
[7] Dawood M. (1995) Pharmacologic stimulation of uterine contraction. Seminars in Perinatology, 19, 73–83.
[8] (a) Lee EJ, Kim JS, Kim HP, Lee JH, Kang SS. (2010) Phenolic constituents from the flower buds of Lonicera japonica and their
5-lipoxygenase inhibitory activities. Food Chemistry, 120, 134–139; (b) Sakushima A, Coskun M, Maoka T. (1995)
Hydroxybenzoic acids from Boreava orientalis. Phytochemistry, 40, 257–261; (c) Stanikunaite R, Khan SI, Trappe JM, Ross SA.
(2009) Cyclooxygenase-2 inhibitory and antioxidant compounds from the Truffle Elaphomyces granulatus. Phytotherapy Research,
23, 575–578; (d) Wang L, Zhang C, Wang Z, Zhang M, Shao L, Xu L. (2008) Studies on chemical constituents of Dendrobium
crystallinum. Zhongguo Zhong Yao Za Zhi, 33, 1847–1848.
[9] (a) Kui LY, Mao WQ. (1991) Methodology for pharmacology of Chinese herbal medicine. Shanghai Science and Technology,
Shanghai, 493–495; (b) Dejana E, Callioni A, Quintana A, de Gaetano G. (1979) Bleeding time in laboratory animals. II - A
comparison of different assay conditions in rats. Thrombosis Research, 15, 191–197; (c) Yamamoto J, Asada N, Miyatake Y,
Masuya M, Yamashita T, Okamoto U. (1987) New modified activated partial thromboplastin time and prothrombin time methods
using a synthetic chromogenic substrate in combination with diazotization. Thrombosis Research, 46, 225–231; (d) Gutiérrez M,
Fernández AI, Revuelta MP, Cantabrana B, Hidalgo A. (1998) Partial contribution of polyamines to the relaxant effect of 17[alpha]-
estradiol in rat uterine smooth muscle. General Pharmacology, 30, 71–77.
... Syringic acid is a compound of high-value pharmacological potential and has been isolated so far as a natural product only from a few plant species, including Scrophularia frutescens, Quercus infectoria, Blumea riparia and Isatidis indigotica [15,16]. Pharmacological effects of the compound include pro-coagulant activity, cardioprotective and hepatoprotective effects, local anesthetic and sedative activity, anti-obesity, anticancer, anti-endotoxic, anti-microbial and cholinesterase inhibitory effects [15][16][17][18]. ...
... Syringic acid is a compound of high-value pharmacological potential and has been isolated so far as a natural product only from a few plant species, including Scrophularia frutescens, Quercus infectoria, Blumea riparia and Isatidis indigotica [15,16]. Pharmacological effects of the compound include pro-coagulant activity, cardioprotective and hepatoprotective effects, local anesthetic and sedative activity, anti-obesity, anticancer, anti-endotoxic, anti-microbial and cholinesterase inhibitory effects [15][16][17][18]. ...
Article
Full-text available
Rumex abyssinicus Jacq. is a perennial medicinal herb widely used in traditional medicine to treat many diseases. Phytochemi-cals of the plant were isolated using column chromatography and thin layer chromatography techniques. Extract, fractions and pure compounds were screened for antimicrobial activity against sensitive and multi-drug resistant microbes and their cytotoxicity was performed on different cancer cell lines. The mechanism of action of purified helminthosporin as well as the potent fraction containing a mixture of two compounds was assessed. Fraction R7C3 was the most potent antibacterial with the lowest MIC value of 0.12 µg/mL. Helminthosporin was the most potent compound with the lowest MIC value of 1.95 µg/mL. The compound was more potent than the antibiotic chloramphenicol against multi-drug resistant (MDR) bacteria with MIC equal to 16 µg/mL. The fraction and helminthosporin were shown to destroy the cell wall of the yeast and bacteria, and DNA fragmentation effect on the genome of Candida albicans and Bacillus cereus. Helminthosporin was the most cytotoxic compound with IC 50 ˂ 10 µM. Fraction R7C3 showed the most potent cytotoxic effects on all cancer cell lines, with IC 50 ranging from ˂1 to 4.35 ng/mL. Our study is the first report on the mechanism of action of helminthosporin, a potent candidate in the development of new drugs against multi-resistant bacteria and cancer cells. In addition, this study uncovered Rumex abyssinicus as a new source of syringic acid and bis(2-ethyloctyl) phthalate.
... Beyond their antimicrobial and antioxidant properties, some PAs, including p-coumaric acid and vanillic acid, have also been shown to promote blood coagulation activity. (Huang et al., 2010). Thus, PAs have the potential to confer antimicrobial and antioxidant properties to hemostatic agents while aiding in coagulation. ...
... Despite previous reports on the procoagulant properties of some PAs, platelet attachment density was qualitatively lower on PA-containing foams. (Huang et al., 2010;Luo et al., 2017). However, the clear activation of platelets on PA foam surfaces is a promising indication that blood clotting via thrombus formation could be achieved in these materials. ...
Article
Full-text available
The leading cause of trauma-related death before arrival at a hospital is uncontrolled blood loss. Upon arrival at the hospital, microbial infections in traumatic wounds become an additional factor that increases mortality. The development of hemostatic materials with antimicrobial and antioxidant properties could improve morbidity and mortality in these wounds. To that end, phenolic acids (PAs) were successfully incorporated into the network of shape memory polymer (SMP) polyurethane foams by reacting them with isocyanates. Resulting PA-containing SMP foam shape memory properties, antimicrobial and antioxidant activity, and blood and cell interactions were characterized. Results showed that p-coumaric, vanillic, and ferulic acids were successfully incorporated into the SMP foams. The PA-containing SMP foams retained the antimicrobial and antioxidant properties of the incorporated PAs, with ∼20% H2O2 scavenging and excellent antimicrobial properties again E. coli (∼5X reduction in CFUs vs. control foams), S. aureus (∼4.5X reduction in CFUs vs. control foams, with comparable CFU counts to clinical control), and S. epidermidis (∼25–120X reduction in CFUs vs. control foams, with comparable CFU counts to clinical control). Additionally, appropriate thermal and shape memory properties of PA foams could enable stable storage in low-profile secondary geometries at temperatures up to ∼55°C and rapid expand within ∼2 min after exposure to water in body temperature blood. PA foams had high cytocompatibility (>80%), non-hemolytic properties, and platelet attachment and activation, with improved cytocompatibility and hemocompatibility in comparison with clinical, silver-based controls. The incorporation of PAs provides a natural non-antibiotic approach to antimicrobial SMP foams with antioxidant properties. This system could improve outcomes in traumatic wounds to potentially reduce bleeding-related deaths and subsequent infections.
... To determine the PT, the patient's PPP was mixed with Owren Koller buffer (basal) or with orchid extract and fractions in a 2:1 ratio (previously diluted in Owren Koller). The effects of different concentrations of the extract (5,10,15,20,25,30,35,40,45,50,55 and 60 mg/mL) and fractions (1.0, 2.5, 5.0, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28 and 30 mg/mL) at different incubation times (0.5, 1, 2, 5, 10 and 20 min) were evaluated. The resulting extract or fraction with PPP was incubated at 37 °C for 180 s; subsequently, 50 µL of a calcium thromboplastin solution were added to 25 µL of this mixture at 37 °C; the time of the appearance of the fibrin mesh was recorded with a coagulometer (STA-COMPACT, Diagnostica Stago). ...
... It is derived from syringic acid, which is synthesized via shikimic acid pathway in plants. The reported activity for this compound is in prevention of cardiovascular diseases [39,40], as antioxidant [41,42], antiinflammatory, neuro and hepatoprotective [39]. The therapeutic property of syringic acid is attributed to the presence of methoxy groups at positions 3 and 5 of the aromatic ring, which confers its antioxidant capacity [39]. ...
Article
Full-text available
Laelia furfuracea is an endemic orchid from Mexico, with antioxidant activity. The objective of this study was to evaluate the effect of hydroethanolic extract and fractions obtained from the orchid leaves on the clotting times of patients with venous thromboembolic disease (VTD) and to identify their tentative compounds. The anticoagulant activity was evaluated by determining prothrombin (PT), thrombin (TT) and, activated partial thromboplastin (APTT) times. Identification of the compounds was carried out using a chromatographic technique with an ultra-high-performance liquid chromatographic analyzer coupled with electrospray ionization with quadrupole time of flight-mass-mass spectrometry. The extract prolonged the clotting times depending on the concentration-response (5-60 mg / mL); 25 mg/mL prolonged the PT (33.2 ± 2.3 s) and TT (33.1 ± 0.3 s); and APTT (61.8 ± 3.4 s) at a concentration of 15 mg/mL. The main groups tentatively identified were xanthine, carboxylic acid, amino acid, and phenolic compounds. These compounds or the synergy between them prolong clotting times. Laelia furfuracea is an orchid with research potential in the search for new anticoagulant agents.
... On the 16 th day, tail tip was punctured with a scalpel and a drop of blood from the supraorbital vein was collected on a glass slide. The clotting time was recorded between blood collection and fibrin formation [18]. The normal time for CT test is 2 to 6 minutes. ...
... [9] Feeding of Bitter melon oil increases the number of apoptotic cells and reduce cell proliferation activity. [35] Anti-adiposity effect of bitter melon seed oil was reported. [36] OEMCS exhibited biphasic effect on plasma recalcification time. ...
... 40 In addition to their antimicrobial properties, phenolic acids demonstrate procoagulant activity. 41,42 Future work will focus on incorporating procoagulant species, such as phenolic acids, into the NTA−DEG foams to increase their clotting capabilities while maintaining the desired degradation profiles, open pore structures, and flexible mechanical properties. ...
Article
Full-text available
Uncontrolled hemorrhage is the leading cause of preventable death on the battlefield and results in ∼1.5 million deaths each year. The primary current treatment options are gauze and/or tourniquets, which are ineffective for up to 80% of wounds. Additionally, most hemostatic materials must be removed from the patient within <12 h, which limits their applicability in remote scenarios and can cause additional bleeding upon removal. Here, degradable shape memory polymer (SMP) foams were synthesized to overcome these limitations. SMP foams were modified with oxidatively labile ether groups and hydrolytically labile ester groups to degrade after implantation. Foam physical, thermal, and shape memory properties were assessed along with cytocompatibility and blood interactions. Degradation profiles were obtained in vitro in oxidative and hydrolytic media (3% H2O2 (oxidation) and 0.1 M NaOH (hydrolysis) at 37 °C). The resulting foams had tunable, clinically relevant degradation rates, with complete mass loss within 30–60 days. These SMP foams have potential to provide an easy-to-use, shape-filling hemostatic dressing that can be left in place during traumatic wound healing with future potential use in regenerative medicine applications.
Article
Full-text available
Objective To rapidly identify the two morphologies and chemical properties of similar herbal medicines, Blumea riparia and B. megacephala as the basis for chemical constituent analysis. Methods UPLC-Q-Exactive-MS/MS was utilized for profiling and identification of the constituents in B. riparia and B. megacephala. Chemical pattern recognition (CPR) was further used to compare and distinguish the two herbs and to identify their potential characteristic markers. Then, an HPLC method was established for quality evaluation. Results A total of 93 constituents are identified, including 54 phenolic acids, 35 flavonoids, two saccharides, one phenolic acid glycoside, and one other constituent, of which 67 were identified in B. riparia and B. megacephala for the first time. CPR indicates that B. riparia and B. megacephala samples can be distinguished from each other based on the LC–MS data. The isochlorogenic acid A to cryptochlorogenic acid peak area ratio calculated from the HPLC chromatograms was proposed as a differentiation index for distinguishing and quality control of B. riparia and B. megacephala. Conclusion This study demonstrates significant differences between B. riparia and B. megacephala in terms of chemical composition. The results provide a rapid and simple strategy for the comparison and evaluation of the quality of B. riparia and B. megacephala.
Article
Full-text available
A new xanthene, named blumeaxanthene, was isolated from Blumea riparia DC. The structure was elucidated on the basis of the data of NMR, MS, IR.
Article
Full-text available
A novel xanthene (1), blumeaxanthene II, was isolated from the aerial parts of Blumea riparia (Bl.) DC., a Chinese medicinal plant with hemostatic properties, and its structure was determined by extensive spectroscopic analyses. This compound is the first example of a halogenated xanthene from a natural plant, and a biosynthetic pathway was proposed. Blumeaxanthene II was also tested against Bel-7404 cells in vitro but was found to be only weakly cytotoxic.
Article
Menorrhagia is common, affecting 50% of the female population. It is becoming more apparent that haematological causes, many often under-diagnosed, may be responsible. With increasing laboratory sophistication, platelet functional defects, von Willebrand disease (vWD) and clotting factor deficiencies are diagnosed more frequently. Menorrhagia may be the first clinical manifestation of a bleeding disorder therefore timely diagnosis and management are essential especially if surgery is to be considered as a definitive treatment for menorrhagia not responding to medical management.
Article
Aliphatic esters of protocatechuic acid (PA,1), vanillic acid (VA,9) and gallic acid (GA,18) were prepared and their anti-thrombotic effects were evaluated in the mouse model of thrombosis. The aliphatic groups included methyl, ethyl,n-propyl,i-propyl,n-butyl,i-butyl,n-amyl and cyclohexyl.n-Amyl ester of PA (7), i-propyl and cyclohexyl esters of VA (13 and17 respectively) and ethyl ester of GA (20) treatment significantly lowered the death rate and increased the recovery from paralysis due to the thrombotic challenge. From the limited analogs available, it was tentatively concluded that the structural conformation, where carboxy oxygen (=O or-O−) of the carboxyl group (COOH) at C1 and the oxygen function at C3 (either OH or OCH3) are closely situated, is favorable for the esters of PA, VA and GA to be more antithrombotic.
Article
A new guaiacylglycerol ether, threo-guaiacylglycerol-8′-vanillic acid ether, pyrocatechuic acid, pyrocatechuic acid 3-O-β-d-glucoside, gentisic acid, gentisic acid 5-O-β-d-glucoside, vanillic acid and vanillic acid 4-O-β-d-glucoside were identified from fruits of Boreava orientalis. Structural elucidation was carried out on the basis of UV, mass, 1H and 13C NMR spectral data, including 2D shift-correlation and selective INEPT experiments.
Article
1.1. The effects of 17α-estradiol on KCl (60 mM), CaCl2 (30 μM to 10 mM) and vanadate (0.3 mM)-induced contractions in rat uterus have been assayed. Furthermore, the effect of 17α-estradiol on calmodulin-stimulated cAMP-phosphodiesterase activity was also studied.2.2. 17α-estradiol relaxed the tonic contraction induced by KCI (60 mM) in a concentration-dependent way (IC50, 8.3 ± 0.7 μM), and CaCl2 (0.1 to 10 mM) counteracted it.3.3. CaCl2 (30 μM to 10 mM) produced concentration-dependent contraction of rat uterus in a calcium-free medium supplemented with 60 mM of KCI (EC50:0.2 ± 0.01 mM). 17α-estradiol (8 μM) antagonized the contraction induced by CaCI2, increasing the EC50 value up to 0.7 ± 0.1 mM(P<0.01).4.4. 17α-estradiol (0.1 to 1 mM) relaxed in a concentration-dependent way the tonic contraction induced by vanadate in rat uterus incubated in a calcium-free medium and EDTA supplemented. The maximal relaxation achieved with 1 mM of 17α-estradiol was 52.2 ± 2.8%.5.5. 17α-estradiol (1 to 100 μM) did not modify the basal activity of cAMP-phosphodiesterase but inhibited the calcium plus calmodulin stimulated activity. The maximal inhibition achieved was 43 ± 5.4%.6.6. The relaxing effect of 17α-estradiol on KCI (60 mM)-induced tonic contraction was unmodified with the antioestrogen tamoxifen (0.1 and 1 μM), the inhibitor of tirosine kinase (genistein, 10 μM) and the cAMP-dependent protein kinase inhibitor (Rp-adenosine 3', 5'-monophosphothioate, triethylamine salt, 100 μM). However, the effect was antagonized with the inhibitor of transcription (actinomycin D, 5 μg/ml,), the inhibitor of protein synthesis (cycloheximide, 10 and 100 μg/ml), and the inhibitor of ornithine decarboxilase (α-difluoromethyl-ornithine, 10 mM).7.7. Our results suggest that polyamines contribute to the relaxant effect of 17α-estradiol in rat uterine smooth muscle behaving, presumably, as mediators of the transcriptional component involved in the effect of 17α-estradiol.
Article
Thirteen phenolic constituents, luteolin (1), protocatechuic acid (2), caffeic acid (3), flavoyadorinin-B (4), 4,5-dicaffeoylquinic acid (5), luteolin 7-O-β-d-glucopyranoside (7), 3,5-dicaffeoylquinic acid methyl ester (8), methyl chlorogenate (9), quercetin 3-O-β-d-glucopyranoside (10), 3,5-dicaffeoylquinic acid (11), rhoifolin (12), chlorogenic acid (13), and a novel phenolic glucoside benzoate, vanillic acid 4-O-β-d-(6-O-benzoylglucopyranoside) (6), were isolated from the flower buds of Lonicera japonica. Flavoyadorinin-B (4) was isolated for the first time from a Caprifoliaceae plant. The structures of 1–13 were determined on the basis of chemical and spectroscopic evidence. These compounds were screened for their 5-lipoxygenase inhibitory activity. Only luteolin (1) showed significant inhibitory activity against 5-LOX-catalysed leukotriene production.
Article
Severe postpartum hemorrhage remains an important cause of maternal morbidity and mortality. The most common cause of postpartum hemorrhage is uterine atony. For first line management of postpartum hemorrhage adequate blood and fluid replacement is mandatory. Further therapeutic measures consist of a variety of medical interventions and surgical techniques. A management protocol for treatment of massive obstetric hemorrhage should be available at every delivery unit. Medical therapies that have been reported to successfully control postpartum hemorrhage include administration of uterotonics, and more recently tranexamic acid and recombinant activated factor VII.
Article
Content of protocatechuic acid (PA) in eight locally available fresh fruits was analyzed, and the protective effects of this compound in diabetic mice were examined. PA at 1%, 2%, and 4% was supplied to diabetic mice for 8 weeks. PA treatments significantly lowered plasma glucose and increased insulin levels. PA treatments at 2% and 4% significantly lowered plasminogen activator inhibitor-1 activity and fibrinogen level; increased plasma activity of antithrombin-III and protein C; decreased triglyceride content in plasma, heart, and liver; elevated glutathione level and the retention of glutathione peroxidase and catalase activities in heart and kidney. PA treatments at 2% and 4% also significantly lowered plasma C-reactive protein and von Willebrand factor levels and reduced interleukin-6, tumor necrosis factor-alpha, and monocyte chemoattractant protein-1 levels in heart and kidney. These results support that protocatechuic acid could attenuate diabetic complications via its triglyceride-lowering, anticoagulatory, antioxidative, and antiinflammatory effects.