PROTOCOL
EVALUATION OF TEST FORMULATION IN PREPARATION AND
STANDARDIZATION FOR NOOTROPIC
ACTIVITY
1.0 TEST SYSTEM DETAILS:
Species : Mus musculus (Mouse)
Strain : Swiss Albino/C57bl6
Age : 8-10 weeks
Body Wight :
20-25 g
Sex : Male
No. of animals : 10
/Group
Total Animals : Model (50+10 Extra= 60)
2.0 ALLOCATION OF GROUPS:
Model
:
|
Groups |
Treatment |
Dose; ROA |
No. of Animals |
|
G1 |
Normal
Control |
Normal
saline or 0.25% Na-CMC |
10 |
|
G2 |
Negative
Control (Scopolamine
0.4 mg /kg) |
0.25%
Na-CMC |
10 |
|
G3 |
Reference
Drug- Piracetam (Scopolamine
0.4 mg /kg) |
10 mg/kg; p.o. |
10 |
|
G4 |
Test
Formulation-1 (Scopolamine
0.4 mg /kg) |
X1
mg/kg; p.o. |
10 |
|
G5 |
Test
Formulation-1 (Scopolamine
0.4 mg /kg) |
X2
mg/kg; p.o. |
10 |
*The
doses and ROA (Routes of administration) will be decided based on the type of
reference drug
#
10 extra (~20%) animals will be taken extra due to Scopolamine Induction
possibilities of animal mortalities
3.0 METHOD:
MEMORY MODELS:
EXTEROCEPTIVE BEHAVIOUR MODELS
:
1. Elevated Plus Maze Test :
Elevated plus maze served as the exteroceptive behavioural model
(where in stimulus existed outside the body) to evaluate learning and memory in
mice. The procedure,
technique and endpoint for testing memory shall be followed as per the
parameters described by the investigators working in the area of
psychopharmacology, Transfer latencies (TL) shall be determined on
both 8th and 9th days.
2. Morris - Water maze Test :
The Morris water maze is a sensitive model for studying the
cognitive processes in rodents that not only reliably reflects the ability of
the animal to learn the position of the hidden platform but also reflects
exploratory aspects of behaviour. These rapid developments in the field of
animal models of learning and memory processes may hopefully lead to an
improved understanding of the Patho- Physiology of Alzheimer’s disease, and
finally, permit the rational designing of novel therapeutic strategies for
distinct cognitive dysfunctions. This maze represents a more specific test of
spatial memory, not confounded by the working memory effects
INTEROCEPTIVE
BEHAVIOUR MODELS
Scopolamine and aging induced
amnesia :
Scopolamine is a powerful muscarinic
antagonist capable of crossing the blood-brain barrier and acts both peripherally by
blocking the receptors for acetylcholine at the synapse. It impairs memory
storage of new information (short-term memory) and learning acquisition.
Scopolamine at a dose of 0.4 mg/kg is approved to produce cognitive and memory
changes without causing the debilitating peripheral anticholinergic effects.
2.
BIOCHEMICAL PARAMETERS
Brain Acetyl Cholinesterase (AChE) activity :
The half of brain AChE activity will be assessed using the Ellman colourimetry method. Protein estimation was done by using Folin’s method.
4.0 END POINT PARAMETER(S):
·
Clinical observation
·
Feed water consumption
·
Body Weight.
·
Elevated Plus Maze (Transfer
Latency, Closed Arm Entry, Closed Arm Timing)
·
Morris Water Maze (Escape
Latency, Entry in Target Quadrant, Swimming Speed)
·
Brain Acetyl
Cholinesterase (AChE) activity
Histopathological studies
:
·
The brain will be
collected from the animals and half of the brain fixed in 10% formalin, and
subjected to histopathological examination.
5.0 REFERENCE(S):
1.0
Milind Parle and Mani
Vasudevan. Memory enhancing activity of Abna®: An
Indian ayurvedic poly-herbal formulation. J health sci 2007;53:43-53.
2.0
Lanni C, Lenzken SC, Pascal
A,et al. Cognition enhancers between treating and doping the mind. Pharmacol.
Res 2008;57(3):196-13.
3.0
Joshi H, Parle M. Brahmi
rasayana improves learning and memory in mice. Evidence-based Compl Altern
Med 2006;3(1):79-5.
4.0
Joshi H, Parle M. Cholinergic basic
of memory strengthening effect of Foeniculum
vulgare Linn. J Med Food
2006;9(3):413-17.
5.0
Joshi H, Parle M. Evaluation of the antiamnesic effects of Phyllanthus amarus in mice. Colom Med
2007;38(2):132-39.
6.0
Masahiro T,
Takuji y, Seiichilizuka,Sachiko I, Yasushi I and Yashiokase. Ameliorative
effects of yokukansan,a traditional Japanese medicine, on learning and
non-cognitive disturbances in the Tg2576 mouse model of Alzheimer's disease. J
Ethnopharmacol 2009;122:157-62.
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