It will helpful for doing projects in pharmacology

1. 21-Jul-22 1
PHARMACOLOGICAL SCREENING OF TERMINALIA CHEBULA
FRUIT EXTRACTS AGAINST NEURO DEGENERATIVE DISORDERS
IN RODENTS
Dissertation submitted to
CHALAPATHI INSTITUTE OF PHARMACEUTICAL SCIENCES, LAM, GUNTUR
In the Partial Fulfillment of the Requirements
for the award of the Degree of
MASTER OF PHARMACY
(PHARMACOLOGY)
Submitted By
Kakunuri Lakshmi
Y16MPHPY442
Under the Guidance of
Dr.A.Narendra Babu, M. Pharm., Ph.D.,
Professor
DEPARTMENT OF PHARMACOLOGY
CHALAPATHI INSTITUTE OF PHARMACEUTICAL SCIENCES,
LAM, GUNTUR - 522034.
JUNE 2018

2. CONTENTS
 Abstract
 Introduction
 Plant profile
 Plan of work
 Aim and objectives
 Literature review
 Materials and methods
 Results and discussion
 Summary and Conclusion
 References
21-Jul-22 2

3. ABSTRACT
BACKGROUND:
Terminalia chebula Retz (T.chebula) which is a member of the Combretaceae family is
frequently used medicinal herb in Ayurvedic, Unani, and Siddha & Homeopathy system of
medicine. Terminalia chebula is called the ‘King of Medicine’ in Tibet and is always listed at
the top of the list in Ayurvedic Materia Medica due to its extraordinary power of healing.
OBJECTIVE:
To evaluate the Terminalia chebula fruit extracts against neurodegenerative disorders in
rodents.
MATERIALS AND METHODS:
To screen the Terminalia chebula fruit extracts against haloperidol induced catatonia model
in SD rats (Anti-parkinsonism/ catatonia activity), ethanol- induced cognitive impairment &
diazepam induced amnesia (learning and memory activity in Alzheimer's disease).
RESULTS:
For anti-parkinsonism activity scoring of catalepsy by using block method and motor
activity by using actophotometer where as learning and memory activity was performed to
evaluate the effect of Terminalia chebula on alzeimer’s disease by using the 8-arm radial
maze (8-RAM), Morris water maze (MWM)& histopathological studies by comparing with
the control group.
KEYWORDS:
Terminalia chebula Retz, neurodegenerative disorders, anti-parkinsonism activity, learning
and memory activity in alzeimer’s disease & histopathological studies.
21-Jul-22 3

4. INTRODUCTION
Neurodegenerative diseases (NDDs):
 Traditionally defined as disorders with selective loss of neurons and distinct
involvement of functional systems defining clinical presentation.
 Comprehensive biochemical, genetic and molecular pathological examinations have
expanded this definition.
 During the last century, many studies have demonstrated that proteins with altered
physicochemical properties are deposited in the human brain in NDDs.
 Furthermore, not only neurons but glial cells also accumulate these proteins.(1)
 Examples of neurodegenerative diseases are:
Alzheimer's disease,
Parkinson's disease,
Huntington’s disease,
Amyotrophic lateral sclerosis,
Frontotemporal dementia and
Spinocerebellar ataxias.
 These diseases are diverse in their pathophysiology with some causing memory and
cognitive impairments and others affecting a person’s ability to move, speak and
breath.(2)
21-Jul-22 4

5. TAXONOMICALCLASSIFICATION
21-Jul-22 5
Kingdom : Plantae
Division : Magnoliophyta
Class : Magnoliopsida
Order : Myrtales
Family : Combretaceae
Genus : Terminalia
Species : chebula
Botanical name : Terminalia chebula Retz
Common names: Indian walnut, harad, karakkaya, haritaki.
Parts used: Fruits, roots, bark.(3)
Figure 1: Terminalia chebula fruit, leaf and tree

6. PLAN OF WORK
Collection of Terminalia chebula fruits & authenticated
Powdered
Subjected to extraction with polar (water) & non-polar (ethanol) solvents by using
Soxhlet apparatus
Drying
Preliminary phytochemical analysis
Screening for anti-parkinsonism activity & learning and memory activity in Alzeimer’s
disease
Results and discussions
21-Jul-22 6

7. AIM AND OBJECTIVES
AIM:
To evaluate the pharmacological screening of Terminalia chebula fruit extracts against
neurodegenerative disorders in rodents.
OBJECTIVES:
The objectives of present study are:
i. To carry out the polarity based Terminalia chebula extraction using different
solvents.
ii. To carry out the preliminary phytochemical analysis of Terminalia chebula fruit
extracts.
iii. To screen the Terminalia chebula fruit extracts against haloperidol induced
Parkinsonism model (Anti-parkinsonism activity).
iv. To screen the Terminalia chebula fruit extracts against ethanol- induced cognitive
impairment [8-Arm radial maze (8-RAM)], ethanol- induced cognitive impairment and
diazepam induced amnesia (Morris water maze) (Learning and memory activity in
alzeimer’s model).
21-Jul-22 7

8. LITERATURE REVIEW
 Raju D. et al.,(2009) Performed evaluation of anti-ulcer activity of methanolic extract
of Terminalia chebula fruits in experimental rats.
 Jiban Debnath et al.,(2010) reported anticonvulsant activity of ethanolic extract of
fruits of Terminalia chebula on experimental animals.
 V. Maruthappan et al.,(2010) Performed hypolipidemic activity of haritaki (Terminalia
chebula) in atherogenic diet induced hyperlipidemic rats.
 Seo et al.,(2012) studied anti-arthritic and analgesic effect of NDI10218, a
standardized extract of Terminalia chebula, on arthritis and pain model.
 Walia et al., (2012) studied antioxidant efficacy of fruit extracts of Terminalia chebula
prepared by sequential method using TA 102 strain of Salmonella typhimurium.
 Rohini Ahuja et al.,(2013) reported evaluation of anticancer potential of Terminalia
chebula fruits against ehrlich ascites carcinoma induced cancer in mice.
 T. Savitha et al., (2013) performed antimicrobial evaluation of Terminalia chebula
retz.
 Sarwar et al., (2013) studied antioxidant, cytotoxic and analgesic activities of the
methanolic fruit extract of Terminalia chebula retz.
21-Jul-22 8

9.  Valli. S et al., (2013) studied anticryptococcal activity of Terminalia chebula against
clinical and environmental isolates of cryptococcus neoformans.
 Sireeratawong et al .,(2014) evaluated analgesic and anti-inflammatory activities of
the water extract from Terminalia chebula rezt.
 S.Kirubanandan et al.,(2015) evaluated anti–inflammatory and analgesic activities of
methanol extract of Terminalia chebula fruits.
 Eshwarappa et al .,(2016) Performed anti-lipoxygenase activity of leaf gall extracts
of Terminalia chebula (gaertn.) Retz. (combretaceae).
 S. Yakaew et al.,(2016) evaluated ethanol extract of Terminalia chebula fruit protects
against UVB-induced skin damage.
 Jahan et al.,(2016) Performed Study on Synergistic Activity of Ethanolic Leave
Extract of Terminalia chebula in Combination to Azithromycin and Ciprofloxacin
against Various Bacterial Strains
 Venkatesan et al.,(2017) reported antioxidant, antibacterial activities and identification
of bioactive compounds from Terminalia chebula bark extracts.
 Kesharwani et al.,(2017) studied anti-HSV-2 activity of Terminalia chebula Retz
extract and its constituents, chebulagic and chebulinic acids.
21-Jul-22 9

10. MATERIALS AND METHODS
 Collection and Authentication of Plant Material:
The fruits of T. chebula was identified and purchased from local market of
Guntur and authenticated by DR.P.Satyanarayana Raju (Plant taxonomist) of
department of botony and microbiology in Acharya Nagarjuna University, Guntur.
 Preparation of T. chebula Extractions:
 The Terminalia chebula fruits are powdered in a mechanical grinder.
 The collected powder was successively, extracted with water & ethanol by
using Soxhlet apparatus.
 The extraction was carried out for 72 hrs at a temp not exceeding the boiling point
of the solvent.
 Excess solvent was removed by the solvent evaporation to obtained the dry
weight of the plant extracts .
 Animals:
Sprague dawley (SD) rats of either sex.
21-Jul-22 10

11.  Preliminary Phytochemical Screening:
The Preliminary Phytochemical investigation was carried out with both aqueous
& ethanolic extracts of Terminalia chebula fruit for identification of Phytochemical
constituents. Phytochemical tests were carried out by standard methods.(4)
 Experimental Animals:
SD rats of either sex (200-300g) were maintained for 7 days in the animal
house of Chalapathi Institute of Pharmaceutical Sciences, Guntur under standard
conditions temperature (24 ± 10 C), relative humidity (45-55%) and 12:12 light:
dark cycle. The animals were fed with standard rat pellet and water ad libitum. The
animals were allowed to acclimatize to laboratory conditions 48 h before the start of
the experiment. 5 rats/group were used in all sets of experiments.
 Ethical Approval:
All the protocols were approved by Institutional Animal Ethical Committee
(IAEC) and conducted according to Committee for the Purpose of Control and
Supervision of Experimental Animals (CPCSEA) registered no:
1048/PO/Re/S/07/CPCSEA at Department of Pharmacology, Chalapathi Institute of
Pharmaceutical Sciences, Guntur.
21-Jul-22 11

12.  Drugs and Chemicals :-
 For anti-parkinsonism activity:
 Inducing agent- Haloperidol (4mg/kg, p.o)
 Standard drug – Syndopa (10mg/kg, p.o)
 For learning and memory activity in Alzheimer's disease:-
 Inducing agents-
 Ethanol- 20% (Ethanol was prepared as a 20% solution in sterile normal
saline and administered subcutaneously at a dose of 4.5 gm/ kg ),
 Ethanol - 60%(Ethanol was prepared as a 60% solution in distilled water
and administered i.p at a dose of 2.5 mg/ kg).
 Diazepam- were diluted in normal saline and administered i.p at a dose of
(1 mg/kg).
 standard drug- Donepezil hydrochloride (2.5 mg/kg, p.o)
21-Jul-22 12

13. 21-Jul-22 13
Selection of Methods for Different Activities:

14. INVESTIGATION OF ANTIPARKINSONIAN EFFECT
BY USING BLOCK METHOD & MOTOR ACTIVITY BY USING
ACTOPHOTOMETER
Two wooden blocks, one is being 3cm high and other 9cm high. Catalepsy of an
individual rat was measured by a scoring method given below. Severity of
catatonic response was recorded as follows:
Table 1: Scoring for Catatonia
21-Jul-22 14
Stages Description/ Behaviour Score
Stage- I Rat moves normally when placed on the table 0
Stage- II Rat moves only when touched or pushed 0.5.
Stage -III
Rat placed on the table with front paws set
alternatively on 3cm high block fails to correct the
posture in 10 sec, score=0.5 for each paw with a
total of 1 for this stage.
1
Stage -IV
Rat fails to remove when the front paws are
placed alternatively on 9cm block, score= 1 for
each paw a total score of 2 for this stage.
2

15.  Thus for a single rat, the maximum possible score would be 3.5 revealing total
catatonia. Severity of catatonia was observed at 30, 60, 90, 120 & 240 min after
haloperidol administration. Plot a graph, time along on X-axis and catatonic scores
along the Y-axis. [5]
21-Jul-22 15
Figure-2: Catatonia-Stage-III Figure-3: Catatonia-Stage-IV
Evaluation parameters:
Scoring for catatonia.

16. 21-Jul-22 16
Figure-4: Actophotometer
Evaluation parameters:
 No. of counts / 5 min i.e. after 30 min
 % decrease in motor activity (or) % activity change
=(Before - After 30 minutes / before × 100)

17. Haloperidol Induced Catatonia Model In SD Rats:
Parkinson’s disease was induced by administering haloperidol (4mg/kg p.o) daily for
a week. The SD rats were divided into 4 groups with 5 animals in each group.
Selection of Dose and Treatment Period:
Group-I- Inducing group- Haloperidol (4mg/kg P.O),
Group-II– Standard group- Syndopa plus (10mg/kg P.O) +Haloperidol.
Group-III – Test-1 [Aqueous fruit extract of Terminalia chebula (TCAE-
100mg/kg P.O)] +Haloperidol.
Group-IV – Test-2 [Ethanolic fruit extract of Terminalia chebula (TCEE-
100mg/kg P.O)] +Haloperidol.
All the treatment group animals received respective inducing, standard and
test treatment 30 minutes prior to the haloperidol administration for 7 days of
experimental period.(6,7)
21-Jul-22 17

18. INVESTIGATION OF LEARNING AND MEMORY ACTIVITY IN
ALZHEIMER’S MODEL BY USING MORRIS WATER MAZE (MWM)
21-Jul-22 18
Figure-5:MWM
Evaluation parameters:
Transfer latency in sec

19. Ethanol- Induced Cognitive Impairment:
Ethanol (60%) is used to induce dementia like condition in the dose 2.5 mg/kg
administered i.p for 15 days. The SD rats were divided into 4 groups with 5 animals in
each group.
Selection of Dose and Treatment Period:
Group I: Inducing Group- Ethanol (2.5 mg/kg was administered i.p for 15 days).
Group II: Standard Group -Donepezil hydrochloride (2.5 mg/kg was
administered orally for 15days) + Ethanol.
Group III: Test-I -Aqueous fruit extract of Terminalia chebula [TCAE- 100mg/kg
was administered orally for 15days) + Ethanol.
Group IV: Test -II -Ethanolic fruit extract of Terminalia chebula [TCEE-
100mg/kg was administered orally for 15 days) + Ethanol.(8,9)
All the treatment group animals received respective inducing, standard and
test treatment 30 minutes prior to the ethanol administration for 15 days of
experimental period.
21-Jul-22 19

20. Diazepam Induced Amnesia:
Diazepam is used to induce amnesia like condition in the dose 1 mg/kg administered
i.p for 8 days. The SD rats were divided into 4 groups with 5 animals in each group.
Selection of Dose and Treatment Period:
Group I: Inducing Group-Diazepam (1mg/kg was administered i.p for 8 days).
Group II: Standard Group-Donepezil hydrochloride (2.5 mg/kg was
administered orally for 8days).
Group III: Test-I- Aqueous fruit extract of Terminalia chebula [TCAE- 100mg/kg
was administered orally for 8days).
Group IV: Test –II- Ethanolic fruit extract of Terminalia chebula [TCEE-
100mg/kg was administered orally for 8 days).(9,10)
All the treatment group animals received respective control, standard and test
treatment 30 minutes prior to the diazepam administration for 8 days of
experimental period.
21-Jul-22 20

21. INVESTIGATION OF LEARNING AND MEMORY ACTIVITY IN
ALZHEIMER’S MODEL BY USING 8-ARM RADIAL MAZE (8-RAM)
21-Jul-22 21
Evaluation parameters:
 No. of entries in baited arms
and non-baited arms.
 Time taken to reach the
paired arm.
 Histopathological Studies
 Weights of different organs.
Figure-6: 8-RAM

22. Ethanol- induced cognitive impairment :
Ethanol (20%) is used to induce dementia like condition in the dose 4.5 gm/kg
administered s.c for 21 days). The SD rats were divided into 4 groups with 5 animals
in each group.
Selection of Dose and Treatment Period:
Group I: Inducing Group - Ethanol (4.5 gm/kg was administered subcutaneously for
21 days).
Group II: Standard Group -Donepezil hydrochloride [8] (2.5 mg/kg was administered
orally for 21 days) + Ethanol.
Group III: Test-I -Aqueous fruit extract of Terminalia chebula [TCAE- 100mg/kg was
administered orally for 21 days) + Ethanol.
Group IV: Test -II -Ethanolic fruit extract of Terminalia chebula [TCEE- 100mg/kg
was administered orally for 21 days) + Ethanol.
All the treatment group animals received respective control, standard and test
treatment 30 minutes prior to the ethanol administration for 21 days of
experimental period. (11)
21-Jul-22 22

23. Histopathological Studies:
At the end of the study the rats were sacrificed on 22nd day by using the
method decapitation and whole brain was collected. Routinely, brain tissue
is fixed in 10% formalin. The brain samples were sending to the embiasis
labs for histopathological studies.
Statistical Analysis:
The values are expressed as mean± SEM. The statistical analysis was performed
using one way analysis of variance (ANOVA) followed by Dunnett’s multiple
comparison test. Comparisons were made between inducing group and test/standard
groups. P-values < or = 0.05 was considered statistically significant. The statistical
analysis was done by using Graph pad prism version no: 6.0.
21-Jul-22 23

24. RESULTS AND DISCUSSION
Table 2: Phytochemical Screening of TCAE & TCEE
21-Jul-22 24
Phytochemical
constituents
Aqueous fruit extract of
Terminalia chebula
Ethanolic fruit extract of
Terminalia chebula
Alkaloids ++ ++
Carbohydrates + +
Flavonoids _ _
Phenols + +
Saponins +++ +++
Terpenoids + +
Sterols + +
Tannins + +
Proteins +++ +++
Amino acids +++ +++
Glycosides ++ ++
Fixed oils and
fatty acids
- -

25. Effect of Fruit Extracts of Terminalia Chebula Retz on Cataleptic Activity:
Haloperidol induced a time dependent increase in cataleptic state in rats, as compared
to vehicle treated groups. Maximum catalepsy score was noted at a time interval of
120-180 min. all the groups i.e. standard (L-dopa + carbidopa), TCAE and TCEE
showed significant (P<0.05) reduction in scores at all time periods. The average scores
for the standard and the test drugs were reduced to that of the Haloperidol group
(Group I).
Table 3: Effect of Fruit Extracts of Terminalia chebula Retz on Haloperidol-Induced
Parkinsonism (Catalepsy)
21-Jul-22 25
S.No Group Treatment
Degree of catatonic response after min
30 60 90 120 240
1. I Haloperidol 0.59±0.06 1.73±0.09 2.72±0.08 2.99±0.03 3.53±0.06
2. II
Standard +
haloperidol
0.31±0.02 0.49±0.02 0.32±0.02 0.31±0.01 0.22±0.02
3. III
TCAE +
haloperidol
0.33±0.02 0.51±0.02 0.53±0.02 0.32±0.01 0.24±0.02
4. IV
TCEE +
haloperidol
0.32±0.02 0.50±0.02 0.32±0.02 0.31±0.02 0.23±0.02

26. 21-Jul-22 26
Figure-7: Effect of Fruit Extracts of Terminalia chebula Retz on Haloperidol-
Induced Parkinsonism (Catalepsy). Values are expressed as Mean ± SEM,
p < 0.05 vs. control (n = 5 animals).
Effect of Fruit Extracts of Terminalia chebula Retz on Behavioural
Parameters (Locomotor Activity):Actophotometer:
Animals treated with haloperidol (4 mg/kg, P.O) alone for 7 days
showed a decrease of locomotor activity [no. of counts / 5 min i.e. after
30 min] on 1st day,4th day &7th day and also reduction in the %change
activity when compared to other groups.

27. 21-Jul-22 27
S.No. Group Treatment
Locomotor activity
[no. of counts / 5 min i.e. after 30 min]
1 ST DAY 4TH DAY 7TH DAY
Before
After
%
Change
activity
Before
After
%
Change
activity
Before
After
%
Change
activity
1. I Haloperidol
266 ±
21.24
150.2 ±
12.43
43.5% 138.8 ±
5.46
74.4 ±
4.95
46.39% 87.0 ±
2.67
49.4 ±
5.04
43.2%
2. II
Standard +
haloperidol
159 ±
6.40
75.0 ±
12.45
52.83% 27.4 ±
2.45
11.4 ±
2.44
58.39% 27.6 ±
1.12
8.0 ±
1.52
71%
3. III
TCAE+
haloperidol
172.6 ±
6.39
86.6 ±
12.92
49.82% 32.4 ±
0.93
21.2 ±
2.33
34.56% 33.0 ±
1.30
21.6 ±
2.25
34.5%
4. IV
TCEE+
haloperidol
165.2 ±
4.81
82.6 ±
12.96
50% 28.4 ±
0.98
17.0 ±
2.47
40.1% 29.0 ±
1.30
12.2 ±
1.93
57.9%
Table 4: Effect of Fruit Extracts of Terminalia chebula Retz on Locomotor Activity

28. 21-Jul-22 28
Figure-8
Figure-9
Figure-8,9: Effect of Fruit
Extracts of Terminalia chebula
Retz on Behavioural
Parameters - Motor Activity.
Values are expressed as mean
± SEM, p < 0.01 vs. control, n =
5 animals.

29. Effect of Fruit Extracts of Terminalia chebula Retz on Behavioural Parameters: i.e.
MWM
Animals treated with ethanol [2.5 mg/kg] alone for15 days showed a increase in
transfer latency in seconds on 1st,7th& 15th days as well as diazepam [1mg/kg] alone for
8 days showed a increase in transfer latency in seconds on 8th day & after 24 hrs i.e. 9th
day.
Table 5:Effect of Fruit Extracts of Terminalia chebula Retz on Ethanol- Induced Cognitive
Impairment
21-Jul-22 29
S.No. Group Treatment
Transfer latency (In seconds)
1st DAY 7th DAY 15th DAY
1. I Ethanol 24.2 ±2.35 27.6±3.89 27.8±4.04
2. II Standard+ethanol 5.6±1.47 4.6±1.21 2.6±0.68
3. III TCAE+ ethanol 7.4±1.69 6.6±2.42 3.2±0.97
4. IV TCEE+ ethanol 5.8±1.62 4.4±1.54 2.8±0.66

30. 21-Jul-22 30
Figure-10: Effect of Fruit Extracts of Terminalia chebula Retz on
Ethanol- Induced Cognitive Impairment. Values are expressed as
Mean ± SEM, p < 0.01 vs. control (n = 5 animals).

31. Table 6: Effect of Fruit Extracts of Terminalia chebula Retz on Diazepam Induced
Amnesia
21-Jul-22 31
S.No. Group Treatment
Transfer latency (In seconds)
8TH DAY
After 24 hours
(i.e.9TH DAY)
1. I Diazepam 26.2±3.89 24.8±4.43
2. II Standard+ diazepam 4.4±0.81 2±0.32
3. III TCAE+ diazepam 9.6±2.40 6.6±1.12
4. IV TCEE+ diazepam 5.6±0.40 2.2±0.80
Figure-11: Effect of Fruit Extracts of
Terminalia chebula Retz on. Values
are expressed as Mean ± SEM, p <
0.05 vs. control (n = 5 animals).

32. Effect of Fruit Extracts of Terminalia chebula Retz on Behavioural Parameters i.e.
8-RAM:
Animals treated with ethanol [4.5 mg/kg] alone for 21 days showed an decrease in
time taken to reach paired arm & number of entries in baited arms and non-baited arms
in 1st,7th , 15th & 21 st days.
Table 7 : Effect of Fruit Extracts of Terminalia chebula Retz on Time Taken to Reach
Paired Arm (Ethanol- Induced Cognitive Impairment)
21-Jul-22 32
S.No. Group Treatment
Time Taken to reach Paired arm (Sec)
1st day 7th day 14th day 21st day
1 I Ethanol 147.4±0.75 138.4±0.93 120.4±2.73 100.4±1.72
2 II
Standard+
ethanol
76.2±0.97 66.6±1.89 57.8±0.86 46.4±2.6
3 III TCAE+ ethanol 127.8±0.86 121.6±0.93 102.2±0.86 76±1.76
4 IV TCEE+ ethanol 103±0.84 96±0.71 89.2±1.43 75.2±1.46

33. 21-Jul-22 33
Figure-12: Effect of Fruit Extracts of Terminalia chebula Retz on
Time Taken to Reach Paired Arm. Values are expressed as Mean ±
SE, p < 0.0001 vs. control (n = 5 animals).

34. Group
Treatme
nt
Number of entries in baited arms and non-baited arm
1st day 7th day 14th day 21st day
B.A N.B.A B.A N.B.A B.A N.B.A B.A N.B.A
I Ethanol 1.4±0.3 1.2±0.2 1.2±0.2 1.6±0.3 1.2±0.2 1.6±0.3 0.8±0.2 0.8±0.2
II
Standard
+ethanol
3.6±0.3 3.4±0.3 3.2±0.4 2.2±0.6 5.8±0.4 8.8±0.4 9.4±0.5 12.8±1.6
III
TCAE+
ethanol
2.2±0.4 2.4±0.4 2.6±0.2 4.4±0.5 5.4±0.5 6.8±0.8 5.8±1.0 7.2±1.6
IV
TCEE+
ethanol
6.4±0.5 7±1.3 7±1.3 11±0.9 10.4±0.9 9±1.0 8.8±1.0 10.2±1.2
21-Jul-22 34
Table 8: Effect of Fruit Extracts of Terminalia chebula Retz on Number of Entries in Baited
Arms and Non-baited Arms (Ethanol- Induced Cognitive Impairment)

35. 21-Jul-22 35
Figure-13: Effect of fruit extracts of Terminalia chebula Retz on number of entries
in baited arms and non-baited arms. Values are expressed as Mean ± SEM, p <
0.0001 vs. control (n = 5 animals).

36. Table 9: Effect of Terminalia chebula Fruit Extracts on the Weights of Different Organs
(Ethanol- Induced Cognitive Impairment)
21-Jul-22 36
Organs
Weight in grams
Inducing
group
Standard
group
Test-I (TCAE) Test-II (TCEE)
Brain 2.22±0.28 2.02±0.01 2.13±0.01 2.02±0.01
Heart 1.42±0.16 0.75±0.01 1.13±0.01 0.7±0.03
Lungs 2.01±0.02 1.62±0.01 1.55±0.01 1.54±0.02
Liver 8.18±0.02 5.43±0.01 6.28±0.03 5.37±0.02
Kidneys 2.37±0.21 1.38±0.03 1.35±0.02 1.05±0.01
Pancreas 1.18±0.03 0.51±0.02 0.65±0.02 0.51±0.01

37. 21-Jul-22 37
Figure-14: Effect of Fruit Extracts of Terminalia chebula retz on
the Weights of Different Organs of Rat Values are expressed as
mean ± SEM, p < 0.05 vs. control (n = 5 animals).

38. 21-Jul-22 38
1 2
3 4
M
M
G
G
Figure-15
1-Inducing group, 2- Standard group, 3-Test-I (TCAE),4-Test-II (TCEE).

39. Figure- 15: Histopathological Changes in Rat Brain Treated with Ethanol (i.e. Inducing
Group), Standard with Ethanol (i.e. Standard group), TCAE (Terminalia chebula Aqueous
Extract i.e. Test-I), TCEE (Terminalia chebula Ethanolic Extract i.e. Test-II) using light
microscopy (Magnification 10 X). (M-Molecular layer, G-Granular layer).
Histological Findings:
The results showed that ethanol significantly increased neuronal death, while the
co treatment of Terminalia chebula with ethanol significantly inhibited the neuronal death
compared to ethanol treated group, which suggest that Terminalia chebula an
antioxidant, may effectively protects against the deleterious effects of ethanol-induced
abnormalities by decreasing neuronal death in rat brain, furthermore the cell counting
under light microscope also showed the same increased neurodegeneration upon
ethanol treated group and decreased significantly when treated with ethanolic extract
Terminalia chebula of as compare to alone ethanol treated group.
21-Jul-22 39

40. SUMMARY AND CONCLUSION
 In the present investigation, Terminalia chebula posses the presence of alkaloids,
carbohydrates, phenols, saponins, terpenoids, sterols, tannins, proteins, amino acids
and glycosides. Terminalia chebula showed anti-cholinergic mechanism i.e. anti-
parkinsonism effect, at an effective dose of 100 mg/kg against haloperidol induced
parkinsonian symptoms. TCEE showed comparatively significant effect exerted than
the standard drug Syndopa in the finding of catalepsy score and locomotor activity.
 Terminalia chebula showed cholinesterase inhibitor mechanism at an effective dose
of 100 mg/kg against ethanol- induced cognitive impairment & diazepam induced
amnesia in rats. TCEE showed comparatively significant effect exerted to standard
drug donepezil hydrochloride in the finding of transfer latency in sec (i.e. learning and
memory activity). This effect is attributed to its ability to improve the levels of the
acetylcholine that are decreased in the Alzheimer’s disease.
21-Jul-22 40

41.  Terminalia chebula showed cholinesterase inhibitor mechanism at an effective dose
of 100 mg/kg against ethanol- induced cognitive impairment. TCEE showed
comparatively significant effect exerted to standard drug donepezil hydrochloride in
the finding of time taken to reach paired arm (sec) & number of entries in baited arms
and non-baited arms (i.e. learning and memory activity). The histopathological study
showed that ethanol induced apoptosis neurodegeneration and the co treatment of
ethanolic extract of Terminalia chebula with ethanol decreased ethanol-induced
apoptotic neurodegeneration in rat brain. This effect is attributed to its ability to
improve the levels of the acetylcholine that are decreased in the Alzheimer’s disease.
 Therefore finally suggests ethanolic extract of Terminalia chebula fruit show same
effects exerted to the standard drugs.
21-Jul-22 41

42. REFERENCES
21-Jul-22 42
1. Gabor G.Kovacs, Neuropathology of Neurodegenerative Diseases: A Practical
Guide, Cambridge University Press, 2015,page no:1.
2. Aaron D. Gitler, Paraminder Dhillonand James Shorter3, Neurodegenerative
disease: models, mechanisms, and a new hope , The Company of Biologists,2017,
page no:499.
3. R.Rathinamoorthy and G.Thilagavathi, Terminalia Chebula - Review on
Pharmacological and Biochemical Studies, International Journal of PharmTech
Research , 2014 Vol.6, page no:98.
4. Manjulika yadav, sanjukta chatterji, sharad kumar gupta and geeta watal,
preliminary phytochemical screening of six medicinal plants used in traditional
medicine , International Journal of Pharmacy and Pharmaceutical Sciences ,
Volume 6, Issue 5, 2014, Pg.no: 540.
5. Hand book of experimental pharmacology, S.K. Kulkarni, 3rd edition,
Pg.no:117,118,142.
6. Sayyed Mateen, Mohib Khan, N. Devanna, M. Farooque, Javed Akhtar Ansari, and
Aziz-ur-rahman, potential antiparkinsonian and antidepressant effects of methanolic
extract of swertia chirata and hemidesmus indicus in wistar rats, European journal
of biomedical and pharmaceutical sciences, Volume 3, Issue 3, 2016 , Pg.no:471.

43. 21-Jul-22 43
7. Sibi P Ittiyavirah, Ruby R, Effect of hydro-alcoholic root extract of Plumbago zeylanica l
alone and its combination with aqueous leaf extract of Camellia sinensis on haloperidol
induced parkinsonism in wistar rats, Annals of Neurosciences, Volume 21, Issue 2, 2014,
Pg.no:48.
8. Sibi P.Ittiyavirah, Reshma Ghosh. Cognitive effects of ethanolic extract of Boerhaavia diffusa
and its silver nanoparticles in ethanolic dementia model. The journal of
phytopharmacology.5 (5); 2016: 185-189. 7.
9. Md. Sahab Uddin, Md. Asaduzzaman, Abdullah Al Mamun, Mohammed Ashraful Iqbal,
Ferdous Wahid and Ram Kamol Rony. Neuroprotective Activity of Asparagus racemosus
Linn. Against Ethanol Induced Cognitive Impairment and Oxidative Stress in Rats Brain:
Auspicious for Controlling the Risk of Alzheimer’s disease. Journal of Alzheimer’s Disease
& Parkinsonism. 6(4); 2016:1-10.
10. Silpa Sundur, Inayat Ali, Dr. A. Venkateshwar Reddy, Dr. D. Satyavathi. Evaluation of anti
depressant and nootropic activity of calophyllum inophyllum. Indo American Journal of
Pharmaceutical Research. 5(05); 2015: 2136-2142.
11. Md. Sahab Uddin, Md. Asaduzzaman, Abdullah Al Mamun, Mohammed Ashraful Iqbal,
Ferdous Wahid and Ram Kamol Rony, Neuroprotective Activity of Asparagus racemosus
Linn. Against Ethanol Induced Cognitive Impairment and Oxidative Stress in Rats Brain:
Auspicious for Controlling the Risk of Alzheimer’s Disease, Journal of Alzheimer’s
Disease & Parkinsonism, Volume 6, Issue 4, 2016, Page no: 2,3.

44. 21-Jul-22 44

45. 21-Jul-22 45

46. 21-Jul-22 46
Publications

47. 21-Jul-22 47
The research work “Terminalia Chebula Retz improve memory and learning in
Alzheimer’s model: (experimental study in rat)”, is being communicated in Research
Journal of Pharmacy and Technology (RJPT) for publication.

48. 21-Jul-22 48

49. 21-Jul-22 49