Experiment No-14, Pharmacology-III practical B.Pharm

1. Anti-inflammatory activity of drugs using
carrageenan induced paw-edema model
Experiment No-14
Dr. Sameer H. Sawant
Sinhgad Institute of Pharmacy, Narhe
Dr. Sameer H. Sawant, SIOP

2. Aim: To study anti-inflammatory activity of the drug using carrageenan
induced paw- edema model
• Reference:
1. Kulkarni S.K., “Handbook of Experimental Pharmacology”. Vallabh
Prakashan, Delhi.
2. Suryawanshi P. Hemant, Patel R. Mukesh and Pawar P. Sunil. " A
practical Book of Pharmacology-II". Nirali Prakashan, Pune. 1st
edition. 2019
Dr. Sameer H. Sawant, SIOP

3. Important video link
• https://www.youtube.com/watch?v=zXQs0HoxPnY&t=387s
• In Hindi
• https://www.youtube.com/watch?v=e5sFWeDP1bQ
• for sub plantar injection
• https://www.youtube.com/watch?v=FbVQ-FMF8ko&t=318s
Dr. Sameer H. Sawant, SIOP

4. Requirements:
• Animal: Rats (150-200 g)
• Drug/ Chemical: Carrageenan and Indomethacine
•Drug preparation:
a) Carrageenans (prepares 1% w/v solution and inject 0.1 ml underneath the plantar
region)
b) Indomethacin (20 mg/Kg, s.c.) prepare stock solution 4 mg/ml. Inject in proportion of
0.5ml/100 g of body wt of the rat.
c) Normal saline injected in proportion of 0.5ml/100 g of body wt of the rat.
• Equipment and Apparatus: Plethysmograph or Digital Plethysmometer, vernier
calipers, syringes, oral feeding tube, etc.
Dr. Sameer H. Sawant, SIOP

5. Introduction/Theory:
• Inflammation is the tissue response to the infection, irritation or
foreign substance
• It is the part of defence mechanism but when it abnormally increases,
it is damaging.
• Aging is also considered to be an inflammatory response
• There are several tissue factors or mechanisms that are known to be
involved in inflammatory reaction such as release of histamine,
Bradykinin and prostaglandins.
Dr. Sameer H. Sawant, SIOP

6. Principle:
• The inflammatory reaction is readily produced in rats in the form of
paw oedema with the help of irritants.
• The substances such as
Carrageenan
Formalin
Bradykinin
Histamine
5- Hydroxytryptamine (5-HT)
Mustard, or egg white
• when injected in the dorsum of foot of the rat, they produce acute paw
oedema within a few min of injection.
Dr. Sameer H. Sawant, SIOP

7. • Carrageenan- induced paw oedema is the most commonly used
method in experimental pharmacology to study anti-inflammatory
activity of the drug.
• Carrageenan is a sulphated polysaccharide obtained from sea weed
(Rhodophyceae).
• It causes the release of inflammatory mediators such as histamine, 5
HT, Bradykinin and Prostaglandin and
• Produces inflammation and oedema too
Dr. Sameer H. Sawant, SIOP

8. Plethysmograph:
• It is a basic mechanical
assembly containing mercury
• The mercury displacing due to
dipping of the paw can be
straightforwardly pursued
from the scale appended to the
mercury section or modifying
the mercury level in the arm
B.
Dr. Sameer H. Sawant, SIOP

9. The Digital Plethysmometer:
• A highly useful tool in the
measurement of small volume changes
This test is typically used to
To Follow the evolution of the
inflammatory response experimentally
induced in rodents
To screen potential anti-inflammatory
or anti-oedema properties of
pharmacological substances
Dr. Sameer H. Sawant, SIOP

10. Procedure:
1. Weigh the animals and number them
2. Make the mark on both the hind paw (right and left) just beyond the tibio-tarsal
junction so that one can ensure that every time the paw is dipped in the mercury
column up to the fixed mark
3. Note the initial paw volume (both right and left) of each rat by mercury
displacement method.
4. Divide the animals in 02 groups (control and Indomethacine-treated group) each
comprising of at least 06 rats.
5. Control group inject saline and to test group inject Indomethacin by s.c. route
Dr. Sameer H. Sawant, SIOP

11. 6. After 30 min, inject 0.1 ml 1% (w/v) carrageenan in plantar region
of left paw of control group as well as Indomethacine treated group.
7. Right paw of each rat will serve as reference non-inflamed paw for
comparison.
8. Note the paw volume of both the legs of control treated with normal
saline and Indomethacine-treated rats at 15, 30, 60 and 120 min of
carrageenan challenge.
.
Dr. Sameer H. Sawant, SIOP

12. 9. Calculate the % difference in right and left paw volumes of each rat
of control and Indomethacine- treated group.
10. Compare the mean % change in paw volume in control and drug
treated rats
11. Express the % oedema inhibited by anti- inflammatory drug-
Indomethacine
Dr. Sameer H. Sawant, SIOP

13. Observation Table
Sr. No. Treatment Drug/ solution
and Dose
Paw volume (ml) as measured by mercury displacement at
0 15 min 30 min 60 min 120
min
R L R L R L R L R L
1.
Control
group
Normal saline
--
2.
3.
4.
5.
Mean
% dif in R & L
1.
Treated
group
Indomethacine
(20 mg/ Kg)
2.
3.
4.
5.
Mean
% dif in R & LDr. Sameer H. Sawant, SIOP

14. Inference:
• Indomethacine showed anti- inflammatory activity against carrageenan- induced paw oedema.
•
Dr. Sameer H. Sawant, SIOP