EFFECT OF HERBAL EXTRACT ON EXPERIMENTAL MODELS OF INFLAMMATORY BOWEL DISEASES

TEST SYSTEM DETAILS:

Species : Rattus norvegicus (Rat)

Strain : Wistar or Sprague Dawley

Age : 8-10 weeks

Body Wight : 200-220 g

Sex : Male

No. of animals : 10 /Group

Total Animals : 6×8= 48

1.0 ALLOCATION OF GROUPS:

Model – 1: INDOMETHACIN INDUCED COLITIS

Groups	Treatment	Dose; ROA	No. of Animals
G1	Normal Control	Normal saline or 0.25% Na-CMC	6
G2	Normal Control + Indomethacin	7.5 mg/kg; s.c.	6
G3	Normal Control+ Indomethacin + Herbal Extract	30 mg/kg; p.o.	6
G4	Normal Control + Indomethacin + Prednisolone	2 mg / kg; p.o.	6

*The doses and ROA (Routes of administration) will be decided based on the type of reference /Inducing drug.

Model – 2: ACETIC ACID INDUCED COLITIS

Groups	Treatment	Dose; ROA	No. of Animals
G1	Normal Control	Normal saline or 0.25% Na-CMC	6
G2	Normal Control + Acetic Acid	2 ml of 4% intrarectally	6
G3	Normal Control + Acetic Acid + Herbal extract	2 ml of 4% intrarectally (30 mg/kg; p.o.)	6
G4	Normal Control + Acetic Acid + Prednisolone	2 ml of 4% intrarectally (2 mg / kg; p.o.)	6

2.0 METHOD:

Methods :

1. INDOMETHCIN INDUCED COLITIS
The study comprises four different groups of six albino wistar rats in each as follows :
Normal or untreated animals;
Control animals receive only indomethacin (7.5 mg/kg, s.c.);
Animals treated with herbal extract and indomethacin;
Prednisolone treated group, which will receive prednisolone (2 mg/kg orally) and indomethacin.
Animals pretreated with herbal extract for 7 days will be administered Indomethacin (7.5 mg/kg,s.c.) on 8th and 9th day of treatment.
Extract will be administered till 11th day. On the 11th day the animals will be sacrificed by cervical dislocation and dissected.
Colon will be taken out to assess inflammation, based on physical parameters, macroscopy and microscopic features.
Quantification of inflammation would be done using biochemical assay (MPO and lipid peroxides).


2. ACETIC ACID INDUCED ULCERATIVE COLITIS
The study comprises four different groups of six albino wistar rats in each as follows :
Normal or untreated animals;
Control animals receive only acetic acid (2 ml of 4% intrarectally);
Animals treated with herbal extract and acetic acid solution;
Prednisolone treated group receive prednisolone (2 mg/kg orally) and acetic acid solution.
Animals will be treated with herbal extract for 7 days.
On the 8th day, overnight fasted animals will be anaesthetized using pentobarbitone sodium and 2 ml of 4% acetic acid solution will be instilled into rectum.
After 48 h animals will be sacrificed by cervical dislocation and dissected to remove colon.
Waste material will be removed from colon and it will be flushed with saline gently.
Inflammation will be assessed based on physical parameters, macroscopy and microscopic features.
Quantification of inflammation would be done using biochemical assay (MPO and lipid peroxides).

3.0 END POINT PARAMETER(S):

· Clinical observation

· Feed water consumption

· Body Weight

· Macroscopic evaluation (Scoring of ulcers)

· Change in colon weight

· Change in Colon Length

· Change in Colon Width

Ø Estimation of biochemical parameters

· Myeloperoxidase (MPO)

· Lipid peroxides

Ø Microscopic evaluation (Histopathology)

4.0 MOA-1 : The mechanisms of action of Indomethacin include inhibition of prostaglandin and leuko-triene synthesis, immunosuppressive activity, free-radical scavenging, impairment of white cell adhesion and function, and inhibition of cytokine synthesis.

MOA-2: Acetic Acid inducing colitis with two origins: Inflammatory cells and tissues damage.

5.0 REFERENCE(S):

3.1 Sellin JH, Pasricha PJ. Pharmacotherapy of inflammatory bowel disease. In: Brunton LL, Lazo JS, Parker KL, editors. Goodman and Gilman’s The Pharmacological Basis of Therapeutics. 11th ed. New York (NY): McGraw Hill; 2006. p. 1009-19.

3.2 Dudzinski DM, Serhan. Pharmacology of eicosanoids. In: Golan DE, Tashjian AH, Armstrong EJ, Armstrong AW, editors. Principles of Pharmacology: The Pathophysiologic Basis of Drug Therapy. 2nd ed. Philadelphia (PA): Lippincott Williams and wilkins; 2008. p. 758.

3.3 Jagtap AG, Shirke SS, Phadke AS. Effect of polyherbal formulation on experimental models of inflammatory bowel diseases. J Ethnopharmacol 2004;90:195-204.

3.4 Bennett PN, Brown MJ. Clinical Pharmacology. 9th ed. New delhi: Elsevier; 2006. p. 645-8.

3.5 Hagar HH, Medany AE, Eter EE, Arafa M. Ameliorative effect of pyrrolidinedithiocarbamate on acetic acid induced colitis in rat. Eur J Pharmacol 2007;554:69-77.

3.6 Dayie NT, Newmen MS, Ayitey-Smith E, Tayman FSK. Screening for antimicrobial activity of Ageratum conyzoides L: A pharmaco-microbiological approach. The internet J Pharmacology 2008;5(2).

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