EVALUATION OF THE EFFICACY OF HERBAL FORMULATIONS IN RAT MODEL OF PHENCYCLIDINE (PCP)-INDUCED SCHIZOPHRENIA

EVALUATION OF THE EFFICACY OF AYURVEDIC FORMULATIONS IN RAT MODEL OF PHENCYCLIDINE (PCP)-INDUCED SCHIZOPHRENIA

1.0 TEST SYSTEM DETAILS:

Species : Rattus norvegicus (Rats)

Strain: Sprague Dawley or Wistar

Age: 6-7 weeks

Sex: Male

No. of animals: 10 /Group

Total animals: 48 Males

2.0 TEST ARTICLES DETAILS

HF1: Herbal Formulation-1

3.0 ALLOCATION OF GROUPS:

Group No.	Group Description	Disease disease-inducing agent administered	Treatment administered	Dose Volume and Route
G1	Normal Control	Sterile normal saline, i.p., q.d., for 14 days	0.5% MC, p.o., b.i.d.	5 ml/kg, p.o.
G2	Disease Control	Phencyclidine (PCP) @ 10 mg/kg/day, i.p. for 14 days	0.5% MC, p.o., b.i.d.
G3	Reference Control		Clozapine: 25 mg/kg, p.o., (Morning) + 0.5% MC (Evening)
G4	Treated with low dose of HB1		HB1: 15-50 mg/kg, b.i.d.  in 0.5% MC
G5	Treated with mid dose of HB1		HB1: 50-150 mg/kg, mg/kg, b.i.d. in 0.5% MC
G6	Treated with high dose 2 of HB1		HB1: 150-500 mg/kg,   b.i.d. in 0.5% MC

Abbreviations: MC-Methyl Cellulose, p.o.-per os. q.d.: quaque die; bid: bis in die

1.0 METHOD:

· Healthy animals will be selected for the study, randomized based on body weight, and will be assigned to 6 groups consisting of 8 animals each.

· Animals of Group G1 will be designated as normal-control and administered 0.5% MC, p.o., b.i.d.

· Disease control animals (assigned to group G2) will receive 0.5% MC, p.o., b.i.d.

· Animals of group G3 will be treated with reference drug clozapine at the dose of 25 mg/kg, p.o., q.d. The treatment will be started 7 days before PCP administration and continued throughout the study period. The animals will additionally receive 0.5% MC, p.o, b.i.d.

· Animals of group G4-G6 will be treated with HB1, at different dose levels ranging from 15-500 mg/kg, b.i.d.

· Normal control group (G1) will be administered sterile normal saline by intraperitoneal route, whereas animals allocated to groups G2 – G6 will be administered PCP @ 10 mg/kg/day by intraperitoneal route for 14 consecutive days.

· Compound administration will be initiated 14 days before PCP injection and continued throughout the study period.

· 24 hours post-PCP injection, behavioural tests will be performed including open-field test and novel object recognition test. Subsequently, three days after the last PCP injection, the animals will be sacrificed under an overdose of thiopentone anesthesia. After the animal dies, the brain of animals will be collected in 10% neutral buffered formalin for histological analysis.

2.0 PARAMETERS TO BE EVALUATED:

· Behavioral test: Open field test and novel object recognition test.

· Histological analysis of brain (Hematoxylin & Eosin-staining and immunohistochemistry).

· Estimation of the levels of Dopamine and its metabolites in the

3.0 REFERENCE(S):

1. Turgeon, S. M., Lin, T., & Subramanian, M. (2007). Subchronic phencyclidine exposure potentiates the behavioral and c-Fos response to stressful stimuli in rats. Pharmacology Biochemistry and Behavior, 88(1), 73-81.

2. Marquis, J. P., Audet, M. C., Doré, F. Y., & Goulet, S. (2007). Delayed alternation performance following subchronic phencyclidine administration in rats depends on task parameters. Progress in Neuro-Psychopharmacology and Biological Psychiatry, 31(5), 1108-1112.

3. Ninan, I., Jardemark, K. E., & Wang, R. Y. (2003). Olanzapine and clozapine but not haloperidol reverse subchronic phencyclidine-induced functional hyperactivity of N-methyl-D-aspartate receptors in pyramidal cells of the rat medial prefrontal cortex. Neuropharmacology, 44(4), 462-472.

4. Shirayama, Y., Yamamoto, A., Nishimura, T., Katayama, S., & Kawahara, R. (2007). Subsequent exposure to the choline uptake enhancer MKC-231 antagonizes phencyclidine-induced behavioral deficits and reduction in septal cholinergic neurons in rats. European Neuropsychopharmacology, 17(9), 616-626.

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