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The Antithrombotic and

Fibrinolytic Activity of the


Aqueous Extract from the
Crude Latex of Ficus pumila
(Moraceae)
Vien Christian L. Lansangan, Kathrizza T. Mabutas,
Rachelle Z. Ong, Arnold M. Perlas,
Jovencio G. Apostol

University of Santo Tomas


Faculty of Pharmacy

Introduction
Clotting

Important Hemostasis mechanism which


prevents excessive blood loss
Platelet Aggregation->Temporary Platelet Plug
Activation of Coagulation Cascade->Stable
fibrin crosslink

Introduction
Clotting

Is harmful when the blood clot/thrombus


dislodges and blocks blood vessels
Brain CV Accident/ Stroke
Lungs Pulmonary embolism

Introduction
Current Treatments
1. Anticoagulants
2.

Fibrinolytic Agents

3.

Anti Platelet Agents

Botanical Description
Kingdom Plantae
Division Magniliophyta
Class
Magniliopsida
Order
Urticales
Family
Moraceae
Genus
Ficus
Species
pumila

(US DANRC, 2011)

Synonyms

Creeping

Fig
Climbing Fig
Fig Ivy

(Quisumbing, 1951)

Morphological Description
Creeping

Fig

prostrate or climbing shrub.


Leaves

Two-ranked
Ovate, 1.5 to 3 cm long
Round or heart shaped base
Entire or wavy margins
On erect branches, 5 to 10 cm

Latex

vascular liquid with rubberlike qualities


Fruits

Large and brown colored


Partially pear-shaped 4 to 6
cm of length
(Quisumbing, 1951)

Objectives
1.
2.

3.

4.

To determine the antithrombotic and


fibrinolytic activity of Ficus pumila.
To determine if the antithrombotic
activity of Ficus pumila is comparable to
that of heparin.
To determine if thefibrinolytic activity of
Ficus pumila is comparable to that of
Streptokinase.
To determine the possible mechanism by
which Ficus pumila prevents thrombosis.

Methodology

Methodology: Preparation of FPAE

(Perello et al., 2000)

Ficus pumila fruit

GUMS
FPAE
layer

INSOLUBLE
PARTICULATES

FPAE
Turbid light-brown solution

Determination of Caseinolytic
Activity of FPAE

(Perello et al., 2000)

In vitro clot dissolution

(Gong et al., 2011)

In Vitro
Clot
Dissolution

Acute Toxicity Study


To

determine doses of FPAE to be


administered.
Based of OECD TG 425 guidelines

Carrageenan-induced tail thrombosis

(Bekemeier et al., 1991; Hagimori et al., 2009)

Tail Thrombosis

Methodology: Statistical Treatment


of Data
All

statistical analyses were performed at


0.05 level of significance.
The Analysis of Variance (ANOVA) was
used to determine whether significant
differences across treatments existed.
Tukeys multiple comparisons test was
utilized to identify significant differences
between specific treatments.

Results and Discussion


Latex Yield
30 fruits weighing 1083.1g yielded
12.0mL of latex
The percentage yield is 1.11%(v/w)
Caseinolytic Activity
FPAE exhibited an average caseinolytic
activity of 0.199Ucas/mL from 2 trials in
the presence of 12mM of cysteine
*Caseinolytic Unit(Ucas) is the amount of enzyme that produces an increase of one absorbance unit
per rninute in the assay conditions

Results and Discussion


Blood Clot Dissolution

Figure 1. Mean Percentage Dissolution across treatment levels


A direct dose-clot dissolution relationship for FPAE was observed. At 100- and
150mg/3mL, the % blood clot dissolution was significant. Streptokinase 10kIU was
statistically proven to be more effective then FPAE in the dissolution of blood clots
after 24h period. 150mg FPAE was equivalent to 52% of 10,000IU of Streptokinase.

Results and Discussion


Prevention of Carrageenan-Induced Tail Thrombosis

Heparin
100IU

Heparin
10IU

FPAE
127mg/kg

FPAE
80mg/kg

FPAE
50mg/kg

Figure 2. Effects of FPAE and Heparin on mice tail Thrombus Lengths


(24 hours after carrageenan administration)

NSS

Figure 3. Mean Length of Thrombosis (over 3-day period)


A direct dose-prevention of tail thrombosis relationship for FPAE was
observed.

Results and Discussion


Prevention of Carrageenan-Induced Tail
Thrombosis
Test Samples

Heparin

FPAE

Doses

Inhibition of thrombosis (%)


24h

48h

72h

100IU

94

92

92

10IU

87

80

80

127mg/kg

60

61

60

80mg/kg

45

50mg/kg

Figure 3. Table of Mean Length of Thrombosis (over 3-day period)


FPAE at 127mg/kg was able to significantly prevent carrageenan-induced
tail thrombosis over the 3 day period. FPAE at 80mg/kg had significant
activity
only on the third day.

Conclusion
FPAE

possesses antithrombotic activity in


vivo as it significantly prevented
carrageenan-induced mice tail thrombosis.
The possible mechanism of action is direct
fibrin hydrolysis by ficin as proven in the
blood clot dissolution test.
Ficin was present in FPAE and had a caseinolytic
activity of 0.199Ucas/mL
FPAE

at 127mg/kg is as effective as 10IU


and 100IU Commercially Available Heparin.
The fibrinolytic activity of 150mg FPAE is
equivalent to 52% of Streptokinase 100kIU

Recommendations
To

determine other possible mechanisms


of action:
platelet aggregation
aPTT and PT

Use

purified ficin instead of crude enzyme


extract.

References
Bekemeier et al. (1985) Carrageenin-induced thrombosis inrats and mice: a
for testing antithrombotic substances? Agents Actions;
16:44651.

model

Gong et al. (2011). Anti-thrombosis effect of diosgenin extract from


Dioscorea
zingiberensis C.H. Wright in vitro and in vivo,
Phytomedicine, Volume 18, Issue 6.
Hagimori et al. (2009). Improving frequency of thrombosis by altering blood flow
in
the carrageenan-induced rat tail thrombosis model. Pharmacological
research the
official journal of the Italian Pharmacological Society, 60(4), 320-323.
Perello, M., Arribere, M. C., Caffini, N. O., & Priolo, N. S. (2000). Proteolytic
Enzymes
from the Latex of Ficus pumila L. (Moraceae). Latin American Journal of Pharmacy , 19
(4), 257-262.
Quisumbing, E. (1951). Medicinal Plants of the Philippines. Manila: Dept. of
Agriculture & Natural Resources.
PLANTS Profile for Ficus pumila. (2011). Retrieved February 16, 2011, from US
Department of Agriculture Natural Resource Conservation Service:http://plants.usda.gov
/java/profile?symbol=FIPU2

THANK YOU!

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