Local
Anaesthetic Activity of Calotropis gigantea
Latex in Frog and Guinea Pig
KK
Rajasekhar*, V Shankarananth, M Rajagowtham, B Gowtham Kumar, B Thejnayan and M Pavankumar
Dept of Pharmaceutical chemistry, Sri Padmavathi School of
Pharmacy,Tiruchanoor, Tirupati-517503, A.P.,
ABSTRACT:
This work evaluated the
local anaesthetic activity of milky latex obtained from Calotropis gigantea
using three different experimental models in frog and Guinea pig. The latex at
dilution
KEY
WORDS: Calotropis
gigantea,milky latex,local anaesthetic activity,nerve block method,muscle
twitch method and infiltration method .
INTRODUCTION:
Calotropis gigantea (Crown flower) is a species of Calotropis,
native to India, Indonesia,
Malaysia,
Philippines,
Thailand,
Sri Lanka,
and China.
It is a large shrub
growing to 4 m tall. It has clusters of waxy flowers that are either white in
colour. Each flower consists of five pointed petals and a small,
elegant "crown" rising from the centre, which holds the stamens. The
plant has oval, light green leaves and milky stem. Calotropis is used as a traditional medicinal plant1
with unique properties2.Traditionally Calotropis is used alone or
with other medicinals3 to treat common disease such as fevers,
rheumatism, indigestion, cough, cold, eczema, asthma, elephantiasis, nausea,
vomiting, diarrohea4. According to Ayurveda, dried whole plant is a
good tonic, expectorant, depurative, and anthelmintic. The dried root bark is a
substitute for ipecacuanha. The root bark is febrifuge, anthelmintic,
depurative, expectorant, and laxative. The powdered root used in asthma,
bronchitis, and dyspepsia. The leaves are useful in the treatment of paralysis,
arthralgia, swellings, and intermittent fevers. The flowers are bitter,
digestive, astringent, stomachic, anthelmintic, and tonic5. Calotropis
is also a reputed homoeopathic drug6.
Calotropis yields a durable fiber
(commercially known as Bowstring of India) useful for ropes, carpets, fishing
nets, and sewing thread. Floss, obtained from seeds, is used for stuffing
purposes. Fermented mixture of Calotropis and salt is used to remove the
hair from goat skins for production of "nari leather" and of sheep
skins to make leather which is much used for inexpensive book-binding7.
Allelopathic effects of Calotropis
on different agricultural crops have not been well studied. Extracts of
different plant parts viz. root, stem, leaf ,and stem+leaf of Calatropis
affect germination and seedling vigor of many agricultural crops have been
reported8. However, extracts of Calatropis failed to produce
any detrimental effects on weeds such as Chenopodium album Melilotus
alba, Melilotus indica, Sphaeranthus indicus, and Phalaris
minor9.
Literature
survey revealed that the latex of calotropis procera10 and
Tabernaemontana coronaria11 exhibited local anaesthetic activity in
frog (nerve block anaesthesia and muscle twitch method) and Guinea pig(infiltration method), and no
researcher has yet reported local anaesthetic activity of Calotropis gigantea.
Therefore, it is worth conducting an investigation on the local
anaesthetic activity of C. gigantea
latex.
Local anaesthetics are the agents which prevent or relieve
pain by interrupting nerve conduction. They bind to a specific receptor site
within the pore of the sodium channels in nerves and block ion movement through
this pore. Their action is restricted to the site of application and rapidly
reverses upon diffusion from the site of action in the nerve12. They
are administered in variety of routes such as topical anaesthesia, infiltration
anaesthesia, field block anaesthesia, nerve block anaesthesia, intravenous
regional anaesthesia, spinal anaesthesia and epidural anaesthesia, and have
potential to produce deleterious side effects 13-17. The present
study was undertaken to screen the local anaesthetic activity of latex obtained
from Calotropis gigantea.
MATERIALS AND METHODS:
Chemicals and equipments: Milky latex of Calotropis gigantea,
centrifuge, Distilled water,0.1N Hcl, Normal saline(0.65%
Nacl),Xylocaine(0.25w/v),Fixing solution, Hair depleting agent, Frog board,
Frog stand, Hammer, Sharp pins, Plastic tray, Sherrington recording drum,
Drum-cylinder with smoked paper wrapped, Myographic lever, Surgical instruments
and Dissection box.
Animals: Frog (bigger size) and
Methods: Nerve block anaesthesia method and Muscle
twitch method (Frog), Infiltration anaesthesia method (Guinea pig).
Collection of milky latex:
The milky latex was collected usually between
Evaluation of local
anaesthetic activity:
Nerve block anaesthesia18:
Two frogs (150 gm)were decerebrated and the upper part
of the spinal cord was destroyed with the help of pithing needle. Abdomen was cut opened and all the
abdominal organs were removed, to get a pouch(sac) made of abdominal walls. The
sciatic nerve was exposed to standard (Xylocaine) and test drug(
Application of drugs:
Piece of cotton was immersed in 0.2% xylocaine solution
and placed in the abdominal pouch of 1st frog. Similarly, separate
piece of cotton was immersed in
Leg withdrawal action:
The frog board was placed vertically so that the hind
legs of frog hang freely. Right and Left hind legs were immersed in the beakers
containing 0.1 N Hcl and normal saline. Sudden leg withdrawal action was noted
before and after administration of standard and test.
Muscle twitch method:
The frog is sacrificed by euthanasia and gastrenimus
sciatic muscle nerve preparation is dissected out from the animal. It was
placed in the Lucas moist chamber filled with frog’s saline. The knee joint was
fixed to one cork of the chamber while the muscle tendon was tied to the hook
of the lever. The speed of the drum was adjusted to the maximum and the lever
is adjusted in horizontal line and the after load screw adjusted such that it
touches the hook lever. The nerve was placed on the electrodes and to avoid
getting it dried, cotton soaked in saline was placed over it. The lever was
allowed to touch lightly on drum. Its position was adjusted about one inch
above the base. The base line was drawn till a simple muscle curve was
obtained. Position of contact was changed to study the influence (local
anaesthetic activity) of standard (Xylocaine) and test drug 19 (
Table No.1: Nerve block
anaesthesia method: Comparison between standard and test drug for local anaesthetic
activity
|
Standard vs Test (Milky
latex) |
Responses |
|
|
Left (leg) |
Right (leg) |
|
|
Before administration of the standard drug |
+ |
+ |
|
Standard drug (xylocaine) |
|
|
|
0 minute |
-- |
-- |
|
1 minute |
+ |
+ |
|
2 minutes |
+ |
+ |
|
3 minutes |
+ |
+ |
|
4 minutes |
- |
- |
|
5 minutes |
- |
- |
|
6 minutes |
- |
- |
|
7 minutes |
- |
- |
|
8 minutes |
- |
- |
|
9 minutes |
- |
- |
|
10 minutes |
- |
- |
|
11 minutes |
-
|
-
|
|
Before administration of the test drug |
+ |
+ |
|
Test drug |
|
|
|
0 minute |
|
|
|
1 minute |
+ |
+ |
|
2 minutes |
+ |
+ |
|
3 minutes |
+ |
+ |
|
4 minutes |
+ |
+ |
|
5 minutes |
- |
- |
|
6 minutes |
- |
- |
|
7 minutes |
- |
- |
|
8 minutes |
- |
- |
|
9 minutes |
-
|
- |
|
10 minutes |
+ |
+ |
Infiltration Anaesthesia:
Two healthy adult male Guinea pigs were taken and fur
on their back was removed by using depleting agents. The depleted area was
washed with normal saline and allowed to dry. Then the standard (Xylocaine) and
test drug (
RESULTS AND DISCUSSION:
Action of the drug:
0.1N Hcl in beaker was used to find out the anaesthetic
activity of the milky latex extract from Calotropis gigantea.
In Nerve block anaesthesia method, before
administration of the drug, when the acid has been contacted with the hind leg
there was a sudden withdrawal of leg from the beaker in which the 0.1N Hcl was
taken. After administration of the drug, when tested with Hcl, there was
absence of withdrawal of leg from the acid. Hence it was proved that the milky
latex from Calotropis gigantea had local anaesthetic activity. The test drug (
Table No.2: Muscle Twitch method:
Comparison between standard and test drug for local anaesthetic activity
|
Group |
Time in minutes |
Response in cm |
|
Control (Before
administration of the drug) Standard drug
(Xylocaine) NS (reproduced) Test (After
administration of the drug) NS (reproduced) |
0 5 10 15 20 25 30 35 38 5 10 15 20 25 30 |
1.5+ 0.01 1.1+ 0.02 0.9 + 0.03 0.8 + 0.02 0.7 + 0.04 0.6 + 0.01 0.5 + 0.02 0.3 + 0.01 1.4 + 0.01 0.6+0.04 0.5+ 0.02 0.4+0.01 0.3+ 0.01 0.1 + 0.02 1.2 + 0.01 |
Table No.3: Infiltration
method: Comparison between standard and test drug for local anesthetic
activity:
|
Group |
Time in minutes |
Squeak Response |
|
Before administration of the drug Standard drug (Xylocaine) Test(After administration of the drug) |
0 1 5 10 15 20 25 30 35 40 1 5 10 15 20 25 27 30 32 |
+ + - - - - - - - + + - - - - - - - + + |
In Muscle twitch method, control shows height of
response (normal animal) of 1.5 cm and after administration of the test drug,
the height gradually decreased up to 25 minutes. The test drug (
In the Infiltration method, before administration of
the drug, there was response for pin prick up to 1 minute. After administration
of the test drug (
CONCLUSION:
From the above observation, we conclude that the onset
of action of time was 5 minutes for
ACKNOWLEDGEMENT:
The authors are thankful to Smt. P. Sulochana, M.A.,
B.Ed., L.L.B, Chairperson, Dr. D. Ranganayakulu, Principal, Dr. D. Sujatha, B. Pushpakumari
and C. P. Pullaiah, SPSP, Tiruchanoor, Tirupati for providing us facilities and
helping us to carry out this research work.
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Received on 08.02.2010
Accepted on 16.03.2010
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Research J. Pharmacology and
Pharmacodynamics 2(2): March –April 2010: 178 -180