N. M. N.

Alencar1
I. S. T. Figueiredo1
M. R. Vale1
Anti-Inflammatory Effect of the Latex from Calotropis F. S. Bitencurt1
J. S. Oliveira2
procera in Three Different Experimental Models: R. A. Ribeiro1
Peritonitis, Paw Edema and Hemorrhagic Cystitis M. V. Ramos2
Original Paper

Abstract trol the paw edema invoked with dextran stimulation (400 mg),

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Latex from Calotropis procera is widely used in folk medicine as a
rich source of biologically active compounds capable of promot-
ing diverse benefits such as control of dermal fungal infections,
antimicrobial activities and pain relief among other useful prop-
erties. The aim of this work was to characterize the anti-inflam-
matory effect of a non-dialysable protein fraction recovered from
the rubber-free latex using three different experimental models
when administrated intravenously. In vivo neutrophil migration
induced by carrageenin (500 mg) was severely inhibited by doses
of latex proteins reaching maximum inhibition (80 %) at 100 mg/
kg. Paw edema exacerbated by the effect of carrageenin was al-
most completely suppressed after 4 hours and was controlled
within the first hour following latex protein administration.
1144 However, the same latex fraction was completely unable to con-
suggesting that the inhibitory effect of the latex is likely to be
cell-mediated. Iphosphamide-induced vesical edema in mice
was also largely prevented by the latex protein fraction. These re-
sults indicate that an effect similar to that of mesna, the classical
drug used for this purpose, is operative. Our findings suggest that
the sample tested seems to act over a wide spectrum as a novel
anti-inflammatory agent. The results also suggest that the active
molecules are of a proteinaceous nature despite the presence of
numerous secondary metabolites naturally occurring in the C.
procera latex.
Key words
Calotropis procera ´ Asclepiadaceae ´ inflammation ´ iphospha-
mide ´ latex ´ mesna ´ proteins

Introduction Roots, bark and leaves of C. procera are known to satisfactorily

The search for new metabolites or drugs that effectively inter-
vene in infection processes or are capable of modulating specific
pathways in the human metabolism is obviously of great rele-
vance. Plants have been proven to be the primary source of biolo-
gically active natural products as their effectiveness is supported
by their previous exploitation in folk medicine. This is the case
with Calotropis procera, a shrub found in Africa, Asia and South
America. Medicinal applications of C. procera are not a novelty.
combat microbial and fungal infections as well exhibiting mol-
luscicidal and schizoticidal activities, among other documented
properties [1], [2], [3]. The plant constitutively produces abun-
dant latex and this secretion is also reported to possess bacterio-
lytic, insecticidal, analgesic, healing on dermal wounds and anti-
diarrhea properties [1], [4], [5], [6], [7]. Although there is convin-
cing evidence of the potential of C. procera, usually the whole la-
tex is taken as a test sample and the nature of the active molecule
for each specific effect remains unknown. According to the litera-

Affiliation
1
Departamento de Farmacologia e Fisiologia, Universidade Federal do Cearµ, Campus do Pici, Fortaleza-Cearµ,
Brasil
2
Departamento de Bioquímica e Biologia Molecular, Universidade Federal do Cearµ, Campus do Pici, Fortaleza-
Cearµ, Brasil

Correspondence
Prof. Dr. Mµrcio Viana Ramos ´ Departamento de Bioquímica e Biologia Molecular ´
Universidade Federal do Cearµ ´ Campus do Pici ´ Cx. Postal 6033 ´ Fortaleza-Cearµ ´ CEP. 60451±970 ´ Brasil ´
Fax: +55-85-288-9821 ´ E-mail: vramos@ufc.br

Received March 5, 2004 ´ Accepted June 26, 2004

Bibliography
Planta Med 2004; 70: 1144±1149 ´  Georg Thieme Verlag KG Stuttgart ´ New York
DOI 10.1055/s-2004-835842
ISSN 0032-0943
ture latexes are rich sources of non-enzymatic and enzymatic
proteins, non-protein, free amino acids, alkaloids and other sec-
ondary metabolites, i. e., a very complex mixture of small com-
pounds and macromolecules [8], [9], [10]. This highly heteroge-
neous fluid mixed in a poly-isoprene-rubber fraction prevents
and limits efficient protein extraction and isolation. Accordingly,
few proteins have been completely isolated from C. procera latex.
Those that have appear to be pathogenesis-related proteins
known as PR-proteins [10], [11].
When damaged, leaves or green parts release the latex secretion
that has a clingy effect capable of immobilizing insects. The latex
was already shown to prevent insect development, and displays
anti-fungal and anti-bacterial activities, reinforcing the hypo-
thesis of its defensive role [4].
Preliminary results demonstrated that the whole latex from C.
procera has also anti-inflammatory effects [12]. The crude dry la-
tex prepared in an organic extraction medium was determined to
diminish carrageenin-induced rat paw edema. However, the use
of the whole latex to perform experiments severely restricts the
search for individual biologically active molecules. Therefore, the
presence of poly-isoprene (rubber) should be avoided, as it may
be responsible for other undesired effects. The present work
aimed to fractionate the fresh latex of C. procera in order to elim-
inate the rubber fraction and recover its water-soluble protein
fraction through extensive dialysis and lyophilization. Further,
the protein fraction, characterized by SDS-PAGE and mass spec-
trometry was tested for its ability to modulate the inflammatory
process in rats induced by carrageenin and dextran and in mice
induced by iphosphamide in three different experimental mod-
els: peritonitis, paw edema and hemorrhagic cystitis.
Material and Methods
General
Formamide, acrylamide, bisacrylamide and other electrophor-
esis reagents were bought from Sigma Chemical Co. St. Louis,
MO (USA). Iphosphamide (Holoxane) and mesna (Mitexan
200) were from ASTA Medica AG (Germany). Carrageenin was
from BDH Chemicals (England) and Dextran 70 from Pharmacia
(USA). Other reagents like NaCl, acetic acid or methanol were of
analytical grade.
Animals
Male Wistar rats (150 ± 200 g) and male Swiss mice (20 ± 30 g)
grown in the animal house of the Universidade Federal do Cearµ
were comfortably maintained in a temperature-controlled room
with access to water and food ad libitum, until use. For each ex-
perimental test, groups of five individuals were separated and
handled separately. The experimental and handling of animals
were done in accordance with the ethical code for animal experi-
mentation (WHO Chronicle, 1995 [13].
The latex
Healthy plants of Calotropis procera (Ait.) R.Br (Asclepiadaceae)
growing in the field around the beaches of Fortaleza, Brazil were
used as the source of fresh latex of C. procera. The plant material
was identified by Prof. Edson Paula Nunes and the voucher No.
Alencar NMN et al. Anti-inflammatory effect of ¼ Planta Med 2004; 70: 1144 ± 1149
32 663 was deposited at the Prisco Bezerra Herbarium of the Uni-
versidade Federal do Cearµ, Brazil. The latex was collected in dis-
tilled water to give a dilution rate of 1 : 2 (v:v). The mixture was
gently agitated during collection to overcome the natural coagu-
lation effect of the material. Later in the laboratory the samples
were centrifuged at 5,000 ” g during 10 min at room tempera-
ture. The rubber-rich precipitate was taken out and the superna-
tant was exhaustively dialyzed against distilled water at room
temperature and newly centrifuged using the conditions de-
scribed above. The clean, rubber-free supernatant was lyophi-
lized and used to further experiments.
Polyacrylamide gel electrophoresis
The protein fraction obtained by fractionation of the latex was
Original Paper
examined by 12.5 % polyacrylamide gel electrophoresis in the
presence of sodium dodecyl sulfate (SDS) and 2-mercaptoetha-
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nol according to Laemmli [14]. Samples were treated with
0.0625 M tris buffer pH 6.8 containing 2 % SDS and 5 % 2-mercap-
toethanol at 100 8C for 4 min and applied into the gel. The run
was performed at 40 mA at room temperature. Gels were stained
with Coomassie brilliant blue solution in water:acetic acid:me-
thanol (8 : 1:3.5) and destained with the same solution without
the dye.
Mass spectrometry analysis
The protein profile of the latex fraction was also performed by
MALDI-TOF mass spectrometry (MS) using a Vision 2000 TOF
Voyager instrument (Finnigan Mat. Bremen, Germany) equipped
with a 337 nm UV laser.
Peritonitis model
Carrageenin (500 mg) was injected intraperitoneally (i. p.) in 1 mL
of sterile 0.15 M NaCl (saline). Four hours later, animals were sa- 1145
crificed and the peritoneal cavity was washed with 10 mL of sal-
ine containing 5 UI/mL of heparin. The fluid was recovered and
total and differential cells were counted. Doses (0.1 mL) of the
lyophilized latex proteins (0.5 up to 100 mg/kg) in saline were in-
jected by the endovenous (i. v.) route in male Wistar rats 30 min
prior the inflammatory stimuli. Control animals received 0.1 mL
of sterile saline (i. v.). Results are expressed as mean  S.E.M of
the number of cells per microliter of peritoneal fluid similar as
described previously [15]. To make evident the involvement of
latex proteins in the anti-inflammatory effect observed, a similar
manipulation was done with the latex proteins in saline that
were heat treated on a stove at 100 8C for 15, 30 and 60 min pre-
viously and tested using the peritonitis model as described
above.
Paw edema model
Paw edema was induced by carrageenin (500 mg/paw) or dextran
(400 mg/paw) prepared in saline. A volume of 0.1 mL was injected
via the subplantar route into the right hind paw of rats that were
under light anesthesia following ether treatment. Paw volume
was measured immediately before the irritant injection, and at
selected time intervals thereafter with a hydroplethysmometer
[16]. Latex protein fraction in 0.1 mL saline (10 mg/kg) was inject-
ed (i. v.) 30 min before injection of the carrageenin or dextran. In
these experiments, control groups received only sterile saline
(i. v.). Results are expressed according to the increase in paw vol-
ume (mL) calculated by subtracting the basal volume. The area
 under the time-course curve was also calculated using a trape-
zoidal rule and results are expressed as arbitrary units and com-
pared to the control [16].
Hemorrhagic cystitis
Iphosphamide (400 mg/kg) in saline was injected (i. p.) and the
mice were sacrificed 12 hours later. Following an abdominal in-
cision, the bladders were removed, dissected and opened to
eliminate their urinary contents. Latex protein fraction in saline
(30 mg/kg) via i. v. route, mesna (40 mg/kg) i. p. or saline (0.1 mL)
i. v. were injected 30 min before and 4 and 8 hours after iphos-
phamide administration. Mesna was used as the inhibitory con-
trol drug upon cystitis [17]. Control animals received saline i. p.
and i. v. Vesical edema was quantified either by measuring the
Original Paper
increase in bladder wet weight [17] or by the determination of
vesical vascular permeability [17]. The bladder wet weight
expressed as mg is reported as mean  S.E.M./20 g of body weight.
To quantify the vesical vascular permeability, Evans' blue dye
(25 mg/kg) was administered i. v. 1 hour before animal sacrifice.
The bladders were dissected, placed in 1 mL formamide and in-
cubated at room temperature overnight. The optical density of
the extracted dye was recorded spectroscopically at 600 nm and
results are reported as mean  S.E.M. (mg) of Evans' blue/20 g of
body weight. For the macroscopic evaluation the bladders were
examined grossly for edema and hemorrhage according to Gray's
criteria [18]. Edema was considered severe (3+) when fluid was
seen externally and internally in the bladder walls, moderate
(2+) when confined to the internal mucosa, mild (1+) between
normal and moderate, and none (0) as normal. Hemorrhage was
scored as follows: (3+), intravesical clots; (2+), mucosal hemato-
mas; (1+), telangiectasie or dilatation of the bladder vessels and
(0) normal.
1146
Statistical analysis
The results are reported as mean  S.E.M. or as median values
(macroscopic data) of at least 5 determinations. Statistical signif-
icance (p < 0.05) was assessed by analysis of variance (ANOVA)
followed by Bonferroni's test. For macroscopic data, statistical
evaluation was done by Kruskal-Wallis non-parametric analysis
of variance followed by the Mann-Whitney U test.
Results and Discussion
The use of plants in medicine stretches back the earliest stages of
civilization and plant molecules still remain as the main source
of drugs exploited for pharmacological purposes. Active com-
pounds identified in different plant tissues and organs are fre-
quently responsible for the increasing development of new med-
icines [5], [6]. As a result, purified biologically active molecules
naturally occurring in plants are applied to assist patients all
over the world as commercialized biotechnological agents (re-
medy) or as natural extracts (phytotherapy). Remedies are large-
ly accessible in developed countries while natural extracts are
mainly exploited in the undeveloped ones. The plant Calotropis
procera is exploited in folk medicine for many unrelated purpo-
ses mainly in India. The plant seems to be an extraordinary
source of biologically active molecules since its roots, bark,
leaves and latex are known to be very useful in health care. As
cited above, the latex of C. procera is reported to possess many in-
Alencar NMN et al. Anti-inflammatory effect of ¼ Planta Med 2004; 70: 1144 ± 1149
teresting pharmacological properties and we have found among
the native people from the Northeast of Brazil that the latex is
highly efficient at abolishing dermal fungi infection commonly
found in people leaving in areas devoid of sanitation, appropriate
healthcare, and access to clean water. Although folk medicine
does not support an anti-inflammatory property of C. procera,
the whole latex dried and re-suspended in different organic
solvent media as well as water has been proven to display an
anti-inflammatory effect upon the paw edema induced in experi-
mental animals [12]. In the present work we investigated the
potential of its rubber-free abundant latex to prevent or reduce
inflammatory events induced in experimental rats or mice in
three different models of inflammation. To test our hypothesis,
we decided to separate out the latex in order to get its richest
proteinaceous fraction fully devoid of rubber and small metabo-
lites. For this, the fresh crude latex was submitted to exhaustive
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dialyses against distilled water using a cut-off membrane of
12,000 Da. This strategy was useful for discrimination of latex
fractions according to both water solubility and molecular size.
The highly soluble protein fraction was lyophilized and used for
further investigation.
Analysis of the protein pattern on polyacrylamide gel electro-
phoresis and mass spectrometry confirmed the presence of
many proteins with molecular weights ranging of 5,000 up to
50,000 Da. Among them three protein bands with molecular
weights of around 30,000 Da were revealed by electrophoresis
(Fig. 1). These represent the major part of the protein content in
the sample. The presence of proteins with a molecular size esti-
mated to be smaller than 12,000 and not lost during the dialysis
step suggests aggregation since MS or electrophoresis was per-
formed under denaturing conditions. The diversity of proteins
and their high solubility observed confirm that the fractionation
procedure of the whole latex was successfully achieved. Al-
though there are scientific data reporting interesting activities
into latex of C. procera, as far as we are aware, this is the first
time a pharmacological application has been investigated in a
specific fraction of the latex and it should be emphasized that it
was carried out in the absence of rubber.
The water-soluble proteins from C. procera latex were very effec-
tive at preventing the inflammatory process trigged by carragee-
nin, in a dose-dependent manner as measured by the population
of neutrophils in the peritoneal cavity of rats upon stimuli
(Fig. 2). Doses as low as 0.5 mg/kg were capable of overcoming
Fig. 1 Protein pat-
tern of the latex frac-
tion analyzed by poly-
acrylamide gel elec-
trophoresis. Lines A ±
C molecular weight
markers: [phosphory-
lase B, 97.0 kDa, bo-
vine serum albumin,
66.0 kDa, ovalbu-
min, 45 kDa, carbo-
nic anhydrase, 30
kDa, trypsin inhibi-
tor, 20.1 kDa, alpha-lactalbumin, 14.4 kDa]; lines D ± I: protein fraction
from C. procera.
the carrageenin effect. Although the effectiveness of latex pro- Fig. 4 Latex of C.
teins to annul the inflammation was remarkable, the highest procera does not al-
dose tested did not reach the performance of the animals receiv- ter the dextran-in-
duced edemato-
ing saline only. A single dose of latex proteins (30 mg/kg) dis- genic effect in rats.
played inhibitory effects on paw edema stimulated by carragee- The animals were
nin in rats. The overall inhibition exceeded 50 % (Fig. 3B). This treated 30 min be-
tendency was already observed within one hour and was main- fore dextran by in-
traplantar injection
tained until the third hour and the edema was almost absent by
(400 mg/paw), with
the fourth hour (Fig. 3A). On the contrary, water-soluble latex saline (Sal) or latex
proteins did not revert at all the dextran-induced paw edemato- (30 mg/kg) intrave-
genic effect in rats (Fig. 4). It is well documented that inflamma- nously. A: edema
tory events trigged by carrageenin involve cell migration while was measured at 30
min, 1, 2, 3 and 4 h
those produced by dextran administration do not [19]. Thus, it
after the inflamma-
is expected that anti-inflammatory activity of the latex proteins tory challenge and

Original Paper
ought be cell-mediated. expressed as the in-
crease in paw vol-

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ume (mL) above its
basal volume. B: the
Fig. 2 Effect of the area under the time-
i. v. injection of C. course curve was de-
procera latex on the termined using a tra-
carrageenin (Cg)-in- pezoidal rule. Each
duced neutrophil mi- point represents the
gration into rat peri- mean  SEM from
toneal cavities. The five animals. * indicates a significant statistical difference (p < 0.05),
animals were treated compared to the control group (C) (ANOVA ± Bonferroni's test).
i. v. (0.1 mL), with sal-
ine (Sal) or latex (0.5,
2.0, 10.0, 50.0 and It has also been documented that some carbohydrate-binding
100.0 mg/kg). Thirty
proteins from plants, named lectins, produce similar inhibitory
minutes later, carrageenin was injected (500 g/cavity; i. p.). The migra-
tion was evaluated 4 h after Cg injection. Values are mean  S.E.M for effects as observed here on inflammation induced by carragee-
the number of animals used (n = 6). * p < 0.05 indicates statistical dif- nin [15]. However a search for hemagglutinating activity, the
ference compared to Sal and # p < 0.05 compared to control (C) (ANO- most evident characteristic for the presence of lectins, failed in
VA ± Bonferroni's test). our test sample. On the other hand this finding does not discard
the presence of a non-agglutinating lectin in the sample. In fact, 1147
affinity chromatography of the latex proteins on a chitin (N-acet-
ylglucosamine polymer) column gives rise to a specifically
Fig. 3 The inhibi- bound fraction devoid of hemagglutinating activity (MVR, un-
tory effect of latex
published observations). This may corroborate work of Alencar
of C. procera on rat
paw edema induced and co-workers [15] who found that the anti-inflammatory ac-
by intraplantar injec- tivity of an N-acetylglucosamine-binding lectin was specifically
tion of carrageenin meditated by its interaction with residues of N-acetylglucos-
(Cg; 500 mg). The amine in leukocyte recruitment. Further efforts will focus on
animals were treat-
purification of this latex protein and investigate its involvement
ed 30 min before Cg
injection, with saline in the anti-inflammatory potency of the latex studied here.
(Sal) or latex (30 mg/
kg) intravenously. A: The latex proteins were challenged to improve the performance
edema was meas- of animals suffering from hemorrhagic cystitis induced by iphos-
ured at 1, 2, 3 and 4
phamide. Hemorrhagic cystitis is currently a limiting side effect
hours after the in-
flammatory stimuli observed in patients submitted to cyclophosphamide- and iphos-
and expressed as phamide-based oncological therapeutics. Severe edema, ulcera-
the increase in paw tion and hemorrhage are commonly observed [20]. Vesicles of
volume (mL) above mice were examined to determine the extent of lesions provoked
its basal volume. B:
by iphosphamide treatment and compared to those of animals
the area under the
time-course curve pre-treated with doses of latex proteins and mesna (Mitexan
was determined 200) the drug usually chosen to combat side effects of iphospha-
using a trapezoidal mide. Vesical wet weight, vascular permeability and macro-
rule. Each point re- scopic analysis were the parameters evaluated as described in
presents the mean 
the Materials and Methods section. Results in Fig. 5 indicate
SEM from five ani-
mals. * indicates a significant statistical difference (p < 0.05), compar- that iphosphamide-treated mice do develop hemorrhagic cysti-
ed to the control group (C) and # compared to the group treated only tis as shown by the increment of the vesical net weight (Fig. 5A)
with Cg (p < 0.05) (ANOVA ± Bonferroni's test). and by the increment of vascular permeability (Fig. 5B). Never-

Alencar NMN et al. Anti-inflammatory effect of ¼ Planta Med 2004; 70: 1144 ± 1149
 Fig. 5 Comparison Fig. 6 Comparison
of the protective ef- of the protective ef-
fects of mesna and fect of mesna and
latex of C. procera latex of C. procera
upon iphosphamide- upon iphosphamide-
induced hemorrhagic induced hemorrhagic
cystitis in mice. Blad- cystitis in mice ac-
ders were examined cording to visual ob-
to vesical wet weight servation. Images of
(A) and vascular per- bladders: pre-treated
meability (B). Ipho- with saline (left); non-treated (center) and pre-treated with latex pro-
sphamide (400 mg/ teins (30 mg/kg, right).
kg) was injected i. p.
and cystitis evaluat-
ed 12 hours after. Fig. 7 Effect of the
Saline (0.1 mL; i. v.), i. v. injection of Calo-
latex (30 mg/kg; i. v.) tropis procera latex
Original Paper

and mesna (40 mg/ upon the carragee-

kg; i. p.) were admi- nin (Cg)-induced

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nistered 30 minutes neutrophil migration
before and 4 and 8 into rat peritoneal
hours after iphos- cavities. The latex
phamide injection. was previously he-
Control animals (C) ated for 0, 15, 30
received saline i. v. and 60 minutes at
and i. p. Vesical wet 100 8C prior to ad-
weight was meas- ministration. The ani-
ured as mg/20 g body weight and vascular permeability expressed as mals were treated i. v. (0.1 mL), with saline (Sal) or latex (30 mg/kg). Thir-
mg of Evans' blue/mg of bladder. Results are expressed as mean  ty minutes later, carrageenin was injected (500 mg/cavity; i. p.). The mi-
S.E.M. * P < 0.05 compared to saline (Sal) group and # P < 0.05 compar- gration was evaluated 4 h after Cg injection. Values are mean  S.E.M for
ed to control group (C) (ANOVA ± Bonferroni's test). the number of animals used (n = 6). * p < 0.05 indicates statistical dif-
ference compared to Sal and # p< 0.05 compared to control (C) (ANOVA
± Bonferroni's test).

Table 1 Macroscopic analysis of bladders from mice according to are from active proteinaceous molecules, the testing sample
Gray's criteria was submitted to heat treatment prior to administration to ani-
mals in the peritonitis model. The sample heated for 15 minutes
Groups Edema level Hemorrhage level
1148 had its anti-inflammatory effect considerably diminished (62 %).
In samples heated for 30 and 60 minutes the anti-inflammatory
Control 0 (0 ± 0) 0 (0 ± 0)
Iphosphamide 2 (1 ± 3) 3 (2 ± 3)
activity was completely abolished (Fig. 7). Together, the results
Iphosphamide + latex proteins 0 (0 ± 1)* 1 (0 ± 2)* presented here suggest that the biological effects observed are a
Iphosphamide + mesna 0 (0 ± 1)* 0 (0 ± 2)* result of proteins from the latex of C. procera.

Iphosphamide (400 mg/kg; i. p.)-induced macroscopic alterations were evaluated 12 h after
its administration in vehicle (saline), latex (30 mg/kg; i. v.) and mesna (40 mg/kg; i. p.) admi-
nistered 30 minutes before and 4 and 8 hors after iphosphamide injection. The results are re-
ported as median and range (N = 6). * P < 0.05 compared to the iphosphamide group (AN-
OVA ± Bonferroni's test).
theless, the latex proteins successfully promoted considerable
amelioration in the pattern of wet weight and permeability of
vesicles from animals previously treated. This protective effect
was similar, although slightly lower to that of mesna (Fig. 5 and
Table 1). The concentration of the active molecule involved in
this phenomenon could represent a minor part of the latex sam-
ple, so it may be much more potent then mesna which was used
at 40 mg/kg. Further identification and isolation of this molecule
will resolve this. Although further investigation is currently in
progress, the potential of this still unknown molecule is greatly
in evidence. It is striking to observe the pharmacological potency
of the latex proteins fraction under hemorrhagic cystitis-iphos-
phamide induced by macroscopic visualization of bladders.
Images of bladders shown in Fig. 6 illustrate the suppressive ef-
fect of latex proteins upon the side effects caused by iphospha-
mide treatment. Finally, to reinforce our hypothesis that all of
the observed anti-inflammatory activities of this latex fraction
The search for new therapeutics follows a laborious, time-con-
suming protocol and is often expensive. The purification and
chemical and biochemical characterization of new molecules be-
fore they are assayed in different experimental models in order
to establish their potential is an important part of the process. It
is not only of great botanic importance, but also of industrial in-
terest to investigate plant species as a source of molecules with
specific activities. This is the aim of our investigation with the la-
tex produced by Calotropis procera. The study of the anti-inflam-
matory activity of a semi-purified fraction of the whole latex ac-
complishes the first step of the project. Further fractionation of
its proteins based on chromatographic techniques are in pro-
gress and will lead to the identification of the active molecule(s).
In conclusion, the latex of C. procera exhibits a potent anti-in-
flammatory activity over a wide spectrum and this activity
seems to be mediated by proteinaceous molecules.
Acknowledgements
This research is supported by grants from FUNCAP and CNPq
from Brazil and International Foundation for Science (M.V.R.

Alencar NMN et al. Anti-inflammatory effect of ¼ Planta Med 2004; 70: 1144 ± 1149
grant number F-3070 ± 2). The authors are indebted to Dr. Peter
Etchells who critically reviewed the English language of the
manuscript.
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