Grant Proposal - ICMR
Grant proposal
1
Assessment of Neurotransmitters and
Antioxidant levels in the Brain to Screen the
effect of Phyllanthus Amarus on Cognition in
Diabetic Rats
Co - Principal Investigator
Dr. Shyamjith Manikkoth
Associate Professor
Yenepoya Medical College
Principal Investigator
Kevin Joakim Pinto
Lecturer
Yenepoya Medical College
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What is Diabetes Mellitus?
What is Cognition?
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Phyllanthus Amarus
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Review of literature
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Diabetes and cognitive impairment
 Insulin and its receptors play a vital role in memory (Sato KK, 2009).
 Hyperglycemia and insulin deficiency will lead to cognitive impairment
(Landon MB, 2009)
 Loss of memory, information processing speed, Executive function,
attention and motor skill speed (Mendelsohn AB, 2007)
 Degeneration of neurons and reduction in the size of hippocampus
(Beissel BJ, 2011)
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Objectives
To study the effect of Phyllanthus amarus(PAEE) and
Phyllanthus amarus (PAAE)
 On cognitive impairment associated with diabetes
mellitus in Wistar Albino rats.
 On blood sugar level.
 On Neurotransmitters and Antioxidant levels
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Hypothesis
Phyllanthus amarus by its dual role will reduce blood
glucose level and improve memory and cognition
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Methodology
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Materials and methods
STREPTOZOTOCIN
ROSIGLITAZONE
WISTAR ALBINO RATS
PHYLLANTHUS AMARUS
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Extracts
 Ethanolic extract of Phyllanthus amarus (PAEE)
 Aqueous extract of Phyllanthus amarus(PAAE)
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GROUP
CATEGORY
I
Normal control (young rats) (DW)
II
Normal control (aged rats) (DW)
III
Diabetic young rats (Streptozotocin)
IV
Diabetic aged rats (Streptozotocin)
V
Diabetic young rats (Streptozotocin)+ PAEE1
VI
Diabetic aged rats (Streptozotocin)+ PAEE2
VII
Diabetic young rats (Streptozotocin)+ PAAE1
VIII
Diabetic aged rats (Streptozotocin)+ PAAE2
IX
Diabetic young rats (streptozotocin)+ Rosiglitazone1
X
Diabetic aged rats (streptozotocin)+Rosiglitazone2
N=6 [ 3 Male And 3 Female Rats] In each group
DW= Distilled Water
PAEE: Phyllanthus Amarus ethanolic extract
PAAE: Phyllanthus Amarus aqueous extract
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Wistor albino rats
Day 1: Diabetes Induced by Streptozotocin
Day 7: Estimation of Blood Glucose Level
Rats with blood glucose level > 250 mg/dl are considered for the study
Memory assessment after confirmation of diabetes mellitus
4th week
Elevated Plus maze
8th week
Hebb William maze
12th week
Morris Water maze
Investigations: Brain Neurotransmittors and Antioxidants
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Evaluation of Cognitive Functions
 Evelated plus maze test
 Hebb William maze test
 Morris water maze test
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Biochemical Estimations
 Estimation of brain neurotransmitters level by using
Spectrophotometric analysis and paper
chromatographic techniques.
 Determination of antioxidant level by using 5 ,5
dithiobis-2 nitro benzoic acid (DTNB)
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Statistical Analysis
 Unpaired student’s t-test for blood glucose
estimation between test and respective controls
 Paired t-test for between comparisons
 Mann-Whitney non –parametric tests for cognitive
function studies
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Expected outcome and Applicability
The study is expected to provide vital information regarding the
dual property of Phyllanthus amarus on diabetes mellitus induced
cognitive impairment.
If the test compound is found to be effective in preventing
diabetes mellitus induced cognitive impairment, further studies can
be carried to establish its dual property clinically.
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Timeline
S. No.
Activities
1.
Review of
Literature
2.
Tool validation and
designing
3.
Pilot study
4.
Modification of tool
5.
Data collection
6.
Analysis &
interpretation
7.
Report writing
JanMar
I year
April-Jun July-Sep
Oct-Dec
Grant proposal
Jan- Mar
II year
AprilJuly-Sep
Jun
Oct-Dec
19
Budget
Item
Categories
Amount of fund per year
I year
II year
Total
A
1.
Recurring
JRF - 16000x24 months
1,92,000
1,92,000
3,84,000
2.
Consumables
Chemical and drugs
1,00,000
1,00,000
2,00,000
Non recurring
Travel
20,000
20,000
40,000
3,00,000
-
3,00,000
20,000
20,000
40,000
6,32,000
3,32,000
9,64,000
3.
B
Equipment
Morris water maze
Overheads
Grand Total
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