Anti-inflammatory activity of drugs using carrageenan induced paw-edema modelExperiment no 14 -converted

Pharmacology-III practical sem -V B.Pharm

1. Anti-inflammatory activity of drugs using
carrageenan induced paw-edema model
Experiment No-14
Dr. Sameer H. Sawant
SinhgadInstitute 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.,“HandbookofExperimentalPharmacology”.Vallabh
Prakashan,Delhi.
2.SuryawanshiP. Hemant, Patel R. Mukesh and PawarP. Sunil. " A
practical Book of Pharmacology-II". NiraliPrakashan, Pune. 1
st
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 (150-200g)
•Drug/Chemical: Carrageenan and Indomethacine
•Drugpreparation:
a)Carrageenans(prepares 1% w/v solution and inject 0.1 ml underneath the plantar
region)
b)Indomethacin(20 mg/Kg,s.c.) prepare stocksolution4mg/ml. Injectinproportionof
0.5ml/100gofbodywtof the rat.
c)Normal saline injectedinproportionof0.5ml/100gofbodywtof the rat.
•Equipment and Apparatus:Plethysmograph or Digital Plethysmometer, vernier
calipers, syringes, oral feeding tube, etc.
Dr. Sameer H. Sawant, SIOP

5. Introduction/Theory:
•Inflammationisthetissueresponsetotheinfection,irritationor
foreignsubstance
•Itisthepartofdefencemechanismbutwhenitabnormallyincreases,
itisdamaging.
•Agingisalsoconsideredtobeaninflammatoryresponse
•Thereareseveraltissuefactorsormechanismsthatareknowntobe
involvedininflammatoryreactionsuchasreleaseofhistamine,
Bradykininandprostaglandins.
Dr. Sameer H. Sawant, SIOP

6. Principle:
•Theinflammatoryreactionisreadilyproducedinratsintheformof
pawoedemawiththehelpofirritants.
•Thesubstancessuchas
✓Carrageenan
✓Formalin
✓Bradykinin
✓Histamine
✓5-Hydroxytryptamine(5-HT)
✓Mustard,oreggwhite
•wheninjectedinthedorsumoffootoftherat,theyproduceacutepaw
oedemawithinafewminofinjection.
Dr. Sameer H. Sawant, SIOP

7. •Carrageenan-inducedpawoedemaisthemostcommonlyused
methodinexperimentalpharmacologytostudyanti-inflammatory
activityofthedrug.
•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:
•Itisabasicmechanical
assemblycontainingmercury
•Themercurydisplacingdueto
dippingofthepawcanbe
straightforwardlypursued
fromthescaleappendedtothe
mercurysectionormodifying
themercurylevelinthearm
B.
Dr. Sameer H. Sawant, SIOP

9. The Digital Plethysmometer:
•Ahighlyusefultoolinthe
measurementofsmallvolumechanges
Thistestistypicallyusedto
✓ToFollowtheevolutionofthe
inflammatoryresponseexperimentally
inducedinrodents
✓Toscreenpotentialanti-inflammatory
oranti-oedemapropertiesof
pharmacologicalsubstances
Dr. Sameer H. Sawant, SIOP

10. Procedure:
1.Weightheanimalsandnumberthem
2.Makethemarkonboththehindpaw(rightandleft)justbeyondthetibio-tarsal
junctionsothatonecanensurethateverytimethepawisdippedinthemercury
columnuptothefixedmark
3.Notetheinitialpawvolume(bothrightandleft)ofeachratbymercury
displacementmethod.
4.Dividetheanimalsin02groups(controlandIndomethacine-treatedgroup)each
comprisingofatleast06rats.
5.ControlgroupinjectsalineandtotestgroupinjectIndomethacinbys.c.route
Dr. Sameer H. Sawant, SIOP

11. 6.After30min,inject0.1ml1%(w/v)carrageenaninplantarregion
ofleftpawofcontrolgroupaswellasIndomethacinetreatedgroup.
7.Rightpawofeachratwillserveasreferencenon-inflamedpawfor
comparison.
8.Notethepawvolumeofboththelegsofcontroltreatedwithnormal
salineandIndomethacine-treatedratsat15,30,60and120minof
carrageenanchallenge.
.
Dr. Sameer H. Sawant, SIOP

12. 9.Calculatethe%differenceinrightandleftpawvolumesofeachrat
ofcontrolandIndomethacine-treatedgroup.
10.Comparethemean%changeinpawvolumeincontrolanddrug
treatedrats
11.Expressthe%oedemainhibitedbyanti-inflammatorydrug-
Indomethacine
Dr. Sameer H. Sawant, SIOP

13. Observation Table
Sr. No. TreatmentDrug/ solution
and Dose
Paw volume (ml) as measured by mercury displacement at
0 15 min 30 min 60 min120
min
R L R L R L R LRL
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 & L
Dr. Sameer H. Sawant, SIOP

14. Inference:
•Indomethacineshowedanti-inflammatoryactivityagainstcarrageenan-inducedpawoedema.
•
Dr. Sameer H. Sawant, SIOP