PHYTOTHERAPY RESEARCH

Phytother. Res. 25: 1336–1341 (2011)

Published online 17 February 2011 in Wiley Online Library
(wileyonlinelibrary.com) DOI: 10.1002/ptr.3428

Protective Effect of Proteins Derived from the

Latex of Calotropis procera against Inflammatory
Hyperalgesia in Monoarthritic Rats

Vijay L. Kumar,1* Priyanka Chaudhary,2 Marcio V. Ramos,3

Madan Mohan2 and Mayara P. V. Matos3
1
Department of Pharmacology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi, India
2
Department of Plant Molecular Biology, Delhi University South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi 110021, India
3
Departmento de Bioquimica de Biologia Molecular, Universidade Federal do Ceara, Campus do Pici, Cx. Postal 6033, Fortaleza‐Ce Brasil,
CEP 60451‐970, Brazil

Calotropis procera (family: Apocynaceae) is a plant growing in the wild and has been used in the traditional
medicinal system for the treatment of various diseases. The plant produces milky latex that possesses potent
antiinflammatory and analgesic properties. In present study the non‐dialysable protein fraction isolated from the
latex (LP) of this plant was evaluated for its efficacy against inflammation in rats where paw edema was induced
by sub‐plantar injection of carrageenin or monoarthritis was induced by intra‐articular injection of Freund's
complete adjuvant (FCA). The effect of LP was evaluated on edema volume in the paw model and on joint
diameter, stair climbing ability, motility, dorsal flexion pain, levels of oxidative stress markers and joint histology
in arthritis model. The protection afforded by LP was compared with that of standard antiinflammatory drug,
diclofenac (5 mg/kg). LP exhibited a dose‐dependent antiinflammatory effect and produced 32% and 60%
inhibition of paw edema at 10 and 25 mg/kg doses and 12% and 36% inhibition of joint inflammation at 50 and
150 mg/kg doses. The protective effect of LP was associated with normalization of joint functions, histology and
levels of oxidative stress markers in joint tissue. The findings of this study suggest that the protein fraction of
latex of Calotropis procera has the potential to relieve inflammation and pain associated with various arthritic
conditions. Copyright © 2011 John Wiley & Sons, Ltd.

Keywords: Calotropis procera; arthritis; latex proteins; inflammatory hyperalgesia.

aqueous suspension of dried latex (DL) induces inflam-

INTRODUCTION
Calotropis procera Ait, R.Br. (Apocynaceae) is a wild
growing tropical shrub that is well known for its poisonous
properties due to the presence of toxic substances in its
latex. However, the plant has been used in the traditional
medicinal system for the treatment of leprosy, ulcers,
tumors, piles and diseases of the liver, spleen and
abdomen (Kirtikar and Basu, 1935). The fresh or warmed
leaves have been applied on painful rheumatic joints,
swellings, sores and wounds (Wealth of India, 1992).
Experimentally various biological activities have been
demonstrated in different parts of this plant and these
include proteolytic, antimicrobial, larvicidal and anthel-
mintic properties (Atal and Sethi, 1961; Malik and
Chugati, 1979; Ramos et al., 2006; Shivakar and Kumar,
2003). The aerial parts of the plant possess latex in
abundance which has been reported to exhibit medicinal
properties such as antidiarrhoeal, anticancer, antidiabetic,
antiarthritic and antioxidant (Kumar et al., 2001; Choedon
et al., 2006; Roy et al., 2005; Kumar and Roy, 2007, 2009).
The latex has also been reported to produce both
proinflammatory and antiinflammatory effects in various
experimental models. While local administration of an
* Correspondence to: Professor Vijay L. Kumar, Department of
Pharmacology, All India Institute of Medical Sciences, Ansari Nagar,
New Delhi 110029, India.
E-mail: kumarvl98@hotmail.com
Copyright © 2011 John Wiley & Sons, Ltd.
mation, its oral administration inhibits the inflammatory
response (Singh et al., 2000; Kumar and Basu, 1994; Arya
and Kumar, 2005). Recently, the fresh latex has been
subjected to dialysis where the dialysable fraction and the
non‐dialysable protein fraction (latex proteins, LP) have
been shown to exhibit proinflammatory and antiinflam-
matory properties, respectively (Alencar et al., 2006). Both
the DL and LP fractions of the latex also exhibit
antinociceptive properties (Dewan et al., 2000; Soares
et al., 2005). In view of the antiinflammatory and analgesic
properties of the LP fraction, the present study was carried
out to evaluate its efficacy against Freund's complete
adjuvant (FCA) induced inflammatory hyperalgesia in a
well established rat model of monoarthritis (Chillingworth
and Donaldson, 2003). A battery of tests were performed
to evaluate its effect on edema formation, motility, stair
climbing ability, pain associated with dorsal flexion, joint
histology and parameters of oxidative stress.
MATERIALS AND METHODS
Latex protein isolation. The latex was collected from the
aerial parts of the plant, C. procera growing in the vicinity of
Fortaleza, Brazil. The plant was identified by a taxonomist
and a voucher specimen (No. 32663) was deposited at
Prisco Bezerra Herbarium of the Universidade Federal do
Ceará, Brazil. The fresh latex collected in water (1:1, v/v)
Received 02 November 2010
Revised 31 December 2010
Accepted 05 January 2011
 ANTI‐ARTHRITIC EFFECT OF C. PROCERA LATEX PROTEINS
was centrifuged at 5000 × g for 10 min at 10 °C and the
rubber rich pellet was discarded. The supernatant was
dialysed against distilled water using a membrane with a
cut‐off value of 8000 Da. The non‐dialysable material was
centrifuged and the supernatant was lyophilized to obtain
the latex protein fraction (LP). The LP fraction of the latex
comprises multiple protein bands as revealed by SDS
electrophoresis and proteins of high molecular weight
present in this fraction exhibit antiinflammatory properties
(Ramos et al., 2009). The LP fraction was reconstituted in
normal saline (NS) and administered in rats by intraper-
itoneal (i.p.) injection.
Animals. The experiments were carried out in Wistar
rats of either sex weighing 120–150 g and were obtained
from the Central Animal Facility of All India Institute
of Medical Sciences, New Delhi, India. The rats were
housed in cages at ambient temperature and provided
food and water ad libitum. For arthritis related studies
the overnight fasted rats were trained for a week to
climb a staircase with steps at 5, 10 and 15 cm where
water was kept at step 2 and food at step 3. The
experiments were performed according to the guide-
lines of Institutional Animal Ethics Committee follow-
ing due approval (453/IAEC/08).
Paw edema model. Edema was induced in the hind paw
of rats by sub‐plantar injection of 0.1 mL of carrageenin
(0.5%) and the rats were divided into four groups
comprising six animals each. The paw volume was
measured up to a fixed mark on the lateral malleolus
just before and 3 h after injecting carrageenin using a
plethysmometer and the edema volume was calculated
by taking the difference between the two volumes
(Winter et al., 1962). The LP fraction (10 and 25 mg/kg
intraperitoneally, i.p.; LP10 and LP25) and the standard
antiinflammatory drug diclofenac (5 mg/kg orally, D5)
were administered 30 min and 1 h before inducing
inflammation, respectively. The antiinflammatory effect
of the test drugs was expressed as the percent inhibition
with respect to the carrageenin control.
Monoarthritis model. Monoarticular arthritis was in-
duced in rats by intra‐articular injection of 0.1 mL of 0.1%
FCA in the left ankle joint (day 0). The rats were
divided into five groups comprising six animals each and
arthritis was induced in groups II–V. Group I, normal
control (NC); Group II, FCA control; Group III, LP
50 mg/kg i.p. (LP50); Group IV, LP 150 mg/kg i.p.
(LP150); Group V, Diclofenac 5 mg/kg orally (D5). The
rats in Group I and II were given 1 mL NS orally and in
other groups LP and diclofenac were administered
30 min and 1 h before injecting FCA, respectively, and
then daily. The joint diameter and functional para-
meters were recorded before injecting FCA and then
daily for 3 days when the joint inflammation was
maximum (Kumar and Roy, 2009). The animals were
killed to study joint histology and to measure the
biochemical parameters of oxidative stress in the joint
tissue.
Measuring increase in joint diameter. The joint diam-
eter was measured by a micrometer screw gauge on day
0 and then daily for 3 days. The increase in diameter
was calculated by subtracting the day 0 values from the
daily measurements and expressed in millimeters (mm).
Copyright © 2011 John Wiley & Sons, Ltd.
1337
Stair climbing ability (SCA). The trained rats were
allowed to climb a staircase and their climbing ability
was scored 0, 1, 2, 3 if the rats did not climb any step,
climbed onto step 1, step 2 and step 3, respectively
(De Castro Costa et al., 1981).
Motility. The rats were observed for a period of 5 min
and their motility was scored 0, 1 and 2 if the rats
avoided touching the feet to ground, walked with
difficulty with toe touching the ground and walked
easily (De Castro Costa et al., 1981).
Dorsal flexion pain (DFP). The ankle joint of rat was
flexed five times until the toes touched the front of
the leg with an inter‐test interval of 5 s. The pain was
scored as 0, if there was no squeaking or leg with-
drawal; 1, if there was either squeaking or leg with-
drawal; 2, if both squeaking and leg withdrawal were
present, thus giving a maximum score of 10 (De Castro
Costa et al., 1981).
Histological analysis of joint tissue. The arthritic limb
was removed above the stifle joint, degloved and fixed
in 1% formaldehyde in saline. The limbs were decal-
cified, processed for paraffin embedding, sectioned,
stained with hematoxylin‐eosin and the sections were
examined for arthritic changes (Anderson et al., 1996).
Estimation of glutathione and thiobarbituric acid
reactive substances (TBARS). The levels of reduced
glutathione (GSH) and TBARS, an index of malonyl-
dialdehyde (MDA) were measured in the joint tissue by
the method of Ellman (1959) and Ohkawa et al. (1979).
Statistical analysis. The values of edema volume,
increase in joint diameter and biochemical analysis are
expressed as mean ± SEM, and ANOVA followed by
post hoc test (LSD) was used to compare the groups.
The stair climbing ability, motility and dorsal flexion pain
are expressed as median score and Kruskal‐Wallis test
was used to compare the groups. The statistical analysis
was carried out by SPSS program, version 11.5. A value
of p < 0.05 was considered statistically significant.
RESULTS
Effect of LP on edema volume
The sub‐plantar injection of carrageenin produced
edema in the hind paw and the edema volume in this
group was 0.44 ± 0.02 mL. Intraperitoneal administra-
tion of LP at 10 and 25 mg/kg doses produced a dose‐
dependent inhibition of the paw inflammation and the
edema volumes in LP10 and LP 25 groups were
0.30 ± 0.02 and 0.18 ± 0.02 mL, respectively. The inhibi-
tory effect of LP was more pronounced than that of
5 mg/kg dose of diclofenac (0.37 ± 0.01 mL) (Fig. 1).
Effect of LP on increase in joint diameter
Intra‐articular injection of FCA produced joint inflam-
mation with a peak effect occurring on day 3 when the
increase in the diameter of the ankle joint was
Phytother. Res. 25: 1336–1341 (2011)
1338 V. L. KUMAR ET AL.

Effect of LP on motility

The motility of arthritic rats in the control group was

impaired with a median score of 0 against a median
score of 2 in the normal rats. The treatment of arthritic
rats with LP (LP50 and LP150) improved their motility
in a dose‐dependent manner and the median motility
score in LP150 group was comparable to that in the
D5 group (Fig. 2B).

Effect of LP on stair climbing ability (SCA)

Injection of FCA into the ankle joint of trained rats

Figure 1. Effect of LP fraction on carrageenin induced paw edema.
Paw edema was induced in rats by sub‐plantar injection of 0.1 mL
of 0.5% carrageenin and the paw volume was measured at 0 h and
at 3 h. LP and diclofenac were administered before injecting
carrageenin and their antiinflammatory effect was evaluated at
3 h. Results are expressed as mean ± SEM (n = 6). *p < 0.05.
impaired their ability to climb a staircase and a median
score of 1 was obtained in the FCA control group on
day 3 against 3 in the NC group. The treatment with LP
produced a dose‐dependent improvement in the SCA
of arthritic rats and a median score of 2 was obtained in
the LP150 group while the median SCA score in D5
group was 3 (Fig. 2C).

Effect of LP on dorsal flexion pain (DFP)

4.39 ± 0.09 mm against 0.15 ± 0.08 mm in the control
group where NS was injected into the joint. The
treatment of arthritic rats with LP produced a dose‐
dependent decrease in joint inflammation and the
increase in joint diameter in LP50 and LP150 groups
was 3.84 ± 0.16 and 2.79 ± 0.22 mm (12% and 36%
inhibition). The increase in joint diameter in the D5
group was 3.05 ± 0.08 mm (31% inhibition) (Fig. 2A).
Injection of FCA lowered the threshold for pain and a
median score of 9 was obtained on day 3 when the
inflammation was at its peak. The rats in the NC group
on the other hand experienced no pain when the limbs
were flexed (DFP score 0). The treatment with LP
significantly ameliorated pain in the arthritic rats where
a median score of 5 was obtained. The beneficial effect

Figure 2. Effect of LP fraction on increase in joint diameter (A), motility score (B), stair climbing ability score (C) and dorsal flexion pain score
(D) in the monoarthritis model. Arthritis was induced in rats by intra‐articular injection of 0.1 mL of 0.1% FCA. LP and diclofenac were
administered before injecting FCA and then daily for 3 days following which their protective effect on parameters shown in this figure was
evaluated. The results in panel A are given as mean ± SEM (n = 6) and in panels B, C and D are given as box plots where bold lines represent
median values (n = 6), boxes represent interquartile ranges (25th and 75th percentiles). *p < 0.05; **p < 0.01.

Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. 25: 1336–1341 (2011)
 ANTI‐ARTHRITIC EFFECT OF C. PROCERA LATEX PROTEINS
Figure 3. Effect of LP on the levels of GSH (A) and TBARS (B) in joint tissue of arthritic rats. Arthritis was induced by intra‐articular injection
of 0.1 mL of 0.1% FCA. LP and diclofenac were administered before injecting FCA and then daily for 3 days following which the rats were
killed for biochemical analysis of oxidative stress markers in the joint tissue. Results are expressed as mean ± SEM (n = 6). *p < 0.05,
**p < 0.001.
of LP was comparable to that of diclofenac where a
median score of 6 was obtained (Fig. 2D).
Effect of LP on tissue levels of GSH and TBARS
In order to determine the effect of LP on oxidative
stress in arthritic joints, the levels of GSH and TBARS
were measured in the joint tissue. Induction of arthritis
with FCA resulted in a marked reduction in the tissue
levels of GSH and an increase in the levels of TBARS.
The treatment of arthritic rats with LP increased the
levels of GSH and decreased the levels of TBARS as
also observed with diclofenac (Fig. 3).
Histological findings
Microscopic examination of the joint tissue in arthritic
rats on day 3 revealed inflammatory changes with influx
of neutrophils and edema. These changes were atten-
uated in LP50 and D5 groups while integrity of the joint
tissue was well preserved in LP150 group (Fig. 4).
DISCUSSION
The latex of the plant Calotropis procera has been
demonstrated to exhibit antiinflammatory and analgesic
properties in various experimental models. The aque-
ous suspension and methanol extract (MeDL) of the
dried latex and non‐dialysable protein fraction of the
latex (LP) have been shown to be effective in inhibiting
edema formation and inflammatory influx of neutrophils
(Arya and Kumar, 2005; Alencar et al., 2004). The oral
administration of MeDL has recently been demonstrated
to afford protection against FCA induced monoarthritis
in rats (Kumar and Roy, 2009). In the present study, the
LP fraction prepared from the latex was first validated
for its antiinflammatory properties in a carrageenin
induced paw edema model. The intraperitoneal admin-
istration of LP produced a dose‐dependent inhibition of
paw inflammation induced by carrageenin and its anti-
inflammatory efficacy was more pronounced than that of
diclofenac, a standard antiinflammatory drug. Since this
fraction produced a significant antiinflammatory effect
Copyright © 2011 John Wiley & Sons, Ltd.
1339
in the paw edema model, it was further evaluated
for its beneficial effect in FCA induced inflamma-
tory hyperalgesia in rats. The intra‐articular injection
of FCA in rats produced a peak inflammatory re-
sponse on day 3. The treatment of rats with the LP
fraction produced a dose‐dependent inhibition of joint
inflammation, however, the dose of LP required to afford
protection in the arthritis model was higher than the dose
exhibiting an antiinflammatory effect in the paw tissue.
Such a difference in dose could be attributed to the low
intra‐articular concentration of LP due to its protein-
aceous nature and tissue granulation in FCA induced
arthritis (Oates et al., 2006; Kleinau et al., 1994). The
antiinflammatory effect of LP has earlier been attributed
to its ability to inhibit edema formation and neutrophil
influx at the site of inflammation (Alencar et al., 2004).
Neutrophil influx is one of the key features of FCA
induced arthritis and its inhibition by LP may signifi-
cantly contribute to its efficacy in arthritis (Bjursten et al.,
1983). The antiinfiltrative effect of LP was also evident
from the histological analysis of the joint in the present
study. Recently, the antiinflammatory activity of LP has
been attributed to its subfraction LPPI which comprises
multiple proteins of high molecular weight compared
with other subfractions. The LPPI has been shown to be
most effective in inhibiting both leukocyte rolling and
adherence and its inhibitory effect on the inflammatory
response involves modulation of nitric oxide (NO) levels
(Ramos et al., 2009). Besides, the LP fraction of latex
used in the present study has been reported to prevent
the lethal effect of systemic inflammatory response
elicited by Salmonella infection (Lima‐Filho et al., 2010).
The role of various cytokines, NO, reactive oxygen
species, free radicals and lipid peroxidation is well
established in the pathogenesis of arthritis (Szekanecz
et al., 2000; Remans et al., 2005). Treatment of arthritic
rats with the LP fraction maintained oxidative homeo-
stasis in the joint tissue. The antioxidant properties of C.
procera latex have earlier been reported in various
animal models (Kumar and Roy, 2007; Roy et al., 2005).
It is also well known for its free radical scavenging
properties (Mueen Ahmed et al., 2003).
FCA induced inflammatory hyperalgesia in rat is a
well established model for the evaluation of analgesic
activity of drugs. In this model a state of functional
disability is attained at the time of peak inflammatory
response when the animal avoids weight bearing on the
Phytother. Res. 25: 1336–1341 (2011)
1340 V. L. KUMAR ET AL.
Figure 4. Effect of LP on joint histology in arthritic rats. Arthritis was induced by intra‐articular injection of 0.1 mL of 0.1% FCA. LP and
diclofenac were administered before injecting FCA and then daily for 3 days following which the rats were killed for the histological analysis
of the joint. Normal control (A), FCA control showing edema and neutrophilic infiltration (B) and attenuation of inflammatory changes with
D5 (C), LP50 (D) and LP150 (E). (Scale bar = 100 μm).
inflamed joint and is unable to freely move and climb
due to pain. Like diclofenac, LP produced improvement
in motility, stair climbing ability and decreased the DFP
score in arthritic rats. Such a protection has been
reported for DL and its methanol extract, MeDL and
could be attributed to its constituents that include
alkaloids, terpenes, flavonoids, lignans and flavanol
glycosides (Kumar and Roy, 2007, 2009; Kumar and
Arya, 2006; Mueen Ahmed et al., 2003). Besides these
phytochemicals, the present study shows that the
proteins derived from latex also exhibit antiarthritic
property that is comparable to that of standard
antiinflammatory drug, diclofenac. Earlier, the LP and
its subfractions have been shown to exhibit antinoci-
ceptive effect that was comparable to that of morphine
(Ramos et al., 2009). Taken together, these studies show
that the latex comprises various constituents that are
effective when administered through different routes.
On the one hand, it possesses orally active constituents
that could be extracted with solvents such as methanol as
reported earlier, while on the other hand, it comprises
REFERENCES
Alencar NM, Figueiredo IS, Vale MR et al. 2004. Anti‐inflammatory
effect of the latex from Calotropis procera in the three
different experimental models: peritonitis, paw edema and
hemorrhagic cystitis. Planta Med 70: 1144–1149.
Alencar NM, Oliveira JS, Mesquita RO et al. 2006. Pro and anti‐
inflammatory activities of the latex from Calotropis procera
(Ait.) R.Br. are triggered by compounds fractionated by
dialysis. Inflamm Res 55: 559–564.
Anderson GD, Hauser SD, Mc Garity KL, Bremer ME, Isakson PC,
Gregory SA. 1996. Selective inhibition of cyclooxygenase
(COX)‐2 reverses inflammation and expression of COX‐2
and interleukin 6 in rat adjuvant arthritis. J Clin Invest 97:
2672–2679.
Copyright © 2011 John Wiley & Sons, Ltd.
proteins exhibiting similar properties but are effective
following parenteral administration (Kumar and Roy,
2007; Alencar et al., 2004).
The results of the present study show that the protein
fraction of the latex ameliorates inflammatory hyper-
algesia and has the therapeutic potential for use in
various arthritic conditions.
Acknowledgements
This work was funded by DST under Indo‐Brazil S&T programme of
Cooperation. M.V.R. is grateful to CNPq, FUNCAP and IFS for
financial support. The Junior Research Fellowship to P.C. from
Department of Biotechnology, New Delhi during the study period is
acknowledged.
Conflict of Interest
The authors have declared that there is no conflict of interest.
Arya S, Kumar VL. 2005. Anti‐inflammatory efficacy of extracts of
latex of Calotropis procera against different mediators of
inflammation. Mediators Inflamm 4: 228–232.
Atal CK, Sethi PD. 1961. Proteolytic activity of some Indian
plants. III. Pharmacological evaluation of calotropin from
Calotropis procera. Indian J Pharm 24: 131–134.
Bjursten LM, Thomsen P, Ahlstedt S, Bagge U. 1983. The kinetics
of leucocyte migration in to rabbit knee joint elicited by
preformed immune complexes with different in vitro char-
acteristics. Immunology 49: 205–213.
Chillingworth NL, Donaldson LF. 2003. Characterisation of a
Freund's complete adjuvant induced model of chronic arthritis
in mice. J Neurosci Methods 128: 45–52.
Phytother. Res. 25: 1336–1341 (2011)
 ANTI‐ARTHRITIC EFFECT OF C. PROCERA LATEX PROTEINS
Choedon T, Mathan G, Arya S, Kumar VL, Kumar V. 2006.
Anticancer and cytotoxic properties of the latex of Calotropis
procera in a transgenic mouse model of hepatocellular
carcinoma. World J Gastroenterol 12: 2517–2522.
De Castro Costa M, De Sutter P, Gybels J, Van HJ. 1981. Adjuvant
induced arthritis in rat: A possible animal model of chronic
pain. Pain 10: 173–185.
Dewan S, Sangraula H, Kumar VL. 2000. Preliminary studies on
the analgesic activity of latex of Calotropis procera. J
Ethnopharmacol 73: 307–311.
Ellman GL. 1959. Tissue sulfhydryl groups. Arch Biochem Biophys
82: 70–77.
Kirtikar KR, Basu BD. 1935. Indian Medicinal Plants. Lolit Mohan
Basu: Allahabad, India.
Kleinau S, Erlandsson H, Klareskog L. 1994. Percutaneous
exposure of adjuvant oil causes arthritis in DA rats. Clin Exp
Immunol 96: 281–284.
Kumar S, Dewan S, Sangraula H, Kumar VL. 2001. Anti‐diarrhoeal
activity of the latex of Calotropis procera. J Ethnopharmacol
76: 115–118.
Kumar VL, Arya S. 2006. Medicinal uses and pharmacological
properties of Calotropis procera. In Recent Progress in
Medicinal Plants, vol. 11, Govil JN (ed). Studium Press:
Houston, TX, 373–388.
Kumar VL, Basu N. 1994. Anti‐inflammatory activity of latex of
Calotropis procera. J Ethnopharmacol 44: 123–125.
Kumar VL, Roy S. 2007. Calotropis procera latex extract affords
protection against inflammation and oxidative stress in
Freund's complete adjuvant‐induced monoarthritis in rats.
Mediators Inflamm 2007: 47523–47529.
Kumar VL, Roy S. 2009. Protective effect of latex of Calotropis
procera in Freund's complete adjuvant induced monoarthritis.
Phytother Res 23: 1–5.
Lima‐Filho JV, Patriota JM, Silva AF et al. 2010. Proteins from
latex of Calotropis procera prevent septic shock due to lethal
infection by Salmonella enterica serovar Typhimurium. J
Ethnopharmacol 129: 327–334.
Malik NN, Chugati MID. 1979. Antimicrobial activity of Calotropis
procera – a preliminary study. Pak J Sci 31: 127–129.
Copyright © 2011 John Wiley & Sons, Ltd.
1341
Mueen Ahmed KK, Rana AC, Dixit VK. 2003. Free radical scavenging
activity of Calotropis species. Indian Drugs 40: 654–655.
Oates KM, Krause WE, Jones RL, Colby RH. 2006. Rheopexy of
synovial fluid and protein aggregation. J R Soc Interface 3:
167–174.
Ohkawa H, Ohishi N, Yagi K. 1979. Assay for lipid peroxides in
animal tissues by thiobarbituric acid reaction. Anal Biochem
95: 351–358.
Ramos MV, Bandeira GP, Freitas CD et al. 2006. Latex constitu-
ents from Calotropis procera (R.Br) display toxicity upon egg
hatching and larvae of Aedes aegypti (Linn.). Mem Inst
Oswaldo Cruz 101: 503–510.
Ramos MV, Oliveira JS, Figueiredo JG et al. 2009. Involvement of
NO in the inhibitory effect of Calotropis procera latex protein
fractions on leukocyte rolling, adhesion and infiltration in rat
peritonitis model. J Ethnopharmacol 125: 387–392.
Remans PH, Oosterhout VM, Smeets TJ et al. 2005. Intracellular
free radical production in synovial T lymphocytes from patients
with rheumatoid arthritis. Arthritis Rheum 52: 2003–2009.
Roy S, Sehgal R, Padhy BM, Kumar VL. 2005. Anti‐oxidant and
protective effect of latex of Calotropis procera against alloxan‐
induced diabetes in rats. J Ethnopharmacol 102: 470–473.
Shivakar YM, Kumar VL. 2003. Anthelmintic activity of latex of
Calotropis procera. Pharm Biol 41: 263–265.
Singh H, Kumar S, Dewan S, Kumar VL. 2000. Inflammation
induced by latex of Calotropis procera – a new model to
evaluate anti‐inflammatory drugs. J Pharmacol Toxicol
Methods 43: 219–224.
Soares PM, Lima SR, Matos SG et al. 2005. Anti‐nociceptive
activity of Calotropis procera latex in mice. J Ethnopharmacol
99: 125–129.
Szekanecz Z, Halloran MM, Volin MV et al. 2000. Temporal
expression of inflammatory cytokines and chemokines in rat
adjuvant induced arthritis. Arthritis Rheum 43: 1266–1277.
Wealth of India. 1992. Raw Materials, vol. 3. Information and
Publication Directorate, CSIR: New Delhi, 78–84.
Winter CA, Risley EA, Nuss GW. 1962. Carrageenin induced
oedema in hind paw of rat in assay for anti‐inflammatory
drugs. Proc Soc Exp Biol Med 111: 544–547.
Phytother. Res. 25: 1336–1341 (2011)