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Abstract
During the past three decades the number of deaths due to CVDs has increased from 15.2% to 28.1% in
India. There are many dietary and lifestyle factors are responsible for this rise. In the common aetiology of
heart diseases stated by acharya Charaka, along with other causes, psychological causes like Chinta
(worry), Bhaya (Fear/Anxiety), manasik trass (mental tension) are mentioned as factors responsible for
heart disease. There is sharp increase in cases of anxiety and depression due to change lifestyle in present
era. Hence, it is essential to evaluate the role of Ayurvedokta psychological factor such as Bhaya (Anxiety)
in the development of heart disease. Chronic unpredictable mild stress (CUMS) is the most elegant model
for evaluation of anxiety in the rats as this model possesses construct, predictive and face validity in rats.
Hence, this model is used in the present study. In CUMS process, animals will be subjected chronically
and unpredictably to a variety of low-grade stressors which resembles to those associated with anxiety
like symptoms in humans and also cause cognition impairment. It is observed that CUMS had generated
the anxiety in rats leading to alteration in normal cardiac physiology.
Keywords – Manasik trass, Bhaya (Anxiety), Hematology Parameters, CVDs
P
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Corresponding author: Dr.Subhash Waghe
Article Info: Published on : 15/10/2025
Evaluation of Effect of Chronic Unpredictable Mild Stress on Albino Rats WSR
To Hematology Parameters
Dr. Subhash Waghe
1
, Dr. Vijay Potdar
2
1
Professor Dept. of Rog Nidan, Sardar Patel Ayurvedic Medical College, Dongariya, Balaghat.
2
Associate Professor Dept. of Rognidana & Vikrutivigyna, Govt. Ayurvedic College, Nagpur.
Cite this article as: - Dr.Subhash Waghe (2025) ; Evaluation of Effect of Chronic Unpredictable Mild Stress on Albino Rats WSR
To Hematology Parameters ;Inter .J. Dignostics and Research 3 (1) 1-8 , DOI : 1 0 . 5 2 8 1 / z e n o d o . 1 7358633
G
A
R
V
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2
Introduction :
Nearly there are 3 million (30 lac) cases of
Myocardial Infarction occurs every year (API
Study) in India and 15 million (1.5 Cr.) cases
across the globe every year. Out of this, 25% are
under 40 age, 50% are under 50 age, 25% > 50
years of age. The death due to myocardial
infarction is increasing in Indian population at an
alarming rate and accounts for around 15-20% of
all deaths. During the past three decades the
number of deaths due to CVDs has increased from
15.2% to 28.1% in India.
[1]
Stress is an important factor having high impact on
the psychological development which alters
emotion, cognition and related behavioral outputs.
Chronic unpredictable mild stress (CUMS) is the
most elegant model for evaluation of anxiety as this
model possesses construct, predictive and face
validity in rats
[2] [3]
. In CUMS process, animals will
be subjected chronically and unpredictably to a
variety of low-grade stressors which resembles to
those associated with anxiety like symptoms in
humans and also cause cognition impairment.
CUMS protocol will be performed in separate room
but the normal animal left unchallenged. During the
7 weeks, animals were submitted to 6 different
stressors: tilted cage (45°), food and water
deprivation, restricted access to food, exposure
to empty bottle, 24 h wet cage (200ml of water in
100g of sawdust bedding), continuous
illumination. These stressors will be randomly
scheduled over a week period and will be repeated
to maintain the aspect of unpredictability.
Hematological parameters like Hb%, RBC count,
WBC count, platelet count are PCV, measured to
know the effect of stressors on hemopoietic
condition.
Review Of Literature :
The number of factors play role in the
development of ischemic heart diseases but over
consumption of oily fatty food and unhealthy
lifestyle (mithya ahar vihar) with mental stress are
the important basic factors enumerated by both the
science. In Ayurveda it can be called as ‘Hrit
Aposhanaj Hrit Roga’ and the pathophysiology of
MI is mentioned by Sushruta in Sutrasthana 15/32
[4]
and Syndrome of MI is mentioned by Sushrut
Uttartantra 43/131-132
[5]
in the form of ‘Hrit
Shoola’. In the common etiology of heart diseases
stated by acharya Charaka, along with other causes,
psychological causes like Chinta (worry), Bhaya
(Fear/Anxiety), manasik trass (mental tension) are
mentioned as factors responsible for heart
disease
[6]
. There is sharp increase in cases of
anxiety and depression due to change lifestyle in
present era. Hence, it is essential to evaluate the
role of Ayurvedokta psychological factor such as
Bhaya (Anxiety) in the development of heart
disease and alteration of normal hematological
indices.
Research Question :
Whether Ayurvedokta Bhaya (fear) acts as a
aetiological factor for development of heart disease
Hypothesis:
Null Hypothesis (H1) -Ayurvedokta
Bhaya (fear) acts as a aetiological factor for
alteration of normal hematological indices.
Alternate Hypothesis (H0)-
Ayurvedokta Bhaya (fear) does not acts as
an aetiological factor for alteration of
normal hematological indices.
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3
Material & Methodology :
Study Design :
Center of Study – Dept of Roga Nidana &
Vikrutvigyana, Government Ayurvedic
College, Nanded And National Testing
Centre, Pune
Study Population And Sampling :
Animal required for the Study
Species/Common name -
Weight - 200-250 g
Gender – Male and Female
Number to be used - 12
Groups : Animals will be divided into 2
groups.
Groups (n = 6)
Treatment
Normal Control
No treatment
Disease Control
Chronic
unpredictable mild
stress induction
Data Collection & Instruments :
The animals will be subjected chronically and
unpredictably to a variety of low-grade stressors
which resembles to those associated with anxiety
like symptoms in humans and also cause cognition
impairment. CUMS protocol will be performed in
separate room. During the 7 weeks, animals will be
submitted to 6 different stressors:
1] tilted cage (45°),
2] Tail Clamping for 3 minutes,
3] Cold swimming for 5 minutes at 4°C
4] exposure to empty bottle,
5] 24 h wet cage,
6] continuous illumination. These stressors will be
randomly scheduled over a one week period and
will be repeated to maintain the aspect of
unpredictability. After confirmation of stress in
animals, Hematological investigations were done.
Assessment Criteria :
1. The normal reference range of HB% in rats is -
10.4 to 16.5 g/dl. The level lower than the
normal range is considered to be significant
decrease.
2. The normal reference range of PCV Value in
rats is 18 to 48% .
3. The normal reference range of RBC Value in
rats is 3.5 to 7.5 10
12
/L or 3.5 to 7.5 x 10
6
/µL.
4. The normal reference range of Platelet Value
in rats is 107 to 61510
9
/L. or 1.7 to 5.57 x
10
6
/µL.
5. The normal reference range of WBC Value in
rats is4.44 to14.8 10
9
/L or 4400 to
14800/cumm.
6. The normal reference range of Neutrophil
count in rats is 13 to 63% or 1.3 to 6.3 10
9
/L.
7. The normal reference range of Lymphocyte
count in rats is 61 to 86 % or 6.1 to 8.6 10
9
/L.
Observation & Result :
Table No. 1 – Hb% Values (G/Dl)
HGB
Group
Animal No
Sex
Marking
g/dL
NC
1
M
1
15.3
2
M
2
17.3
3
M
3
15.8
4
F
1
15.8
5
F
2
16.1
6
F
3
13.8
MEAN
15.7
SD
1.1
DC
7
M
1
16.2
8
M
2
16.6
9
M
3
16.3
10
F
1
16.2
11
F
2
14.5
12
F
3
12.2
MEAN
15.3
SD
1.7
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Table No. 2 – RBC Value (10
12
/L) :
RBC
Group
Animal No
Sex
Marking
*10
12
/L
NC
1
M
1
7.7
2
M
2
8.4
3
M
3
8.2
4
F
1
7.7
5
F
2
8.4
6
F
3
7.1
MEAN
7.9
SD
0.5
DC
7
M
1
8.7
8
M
2
8.6
9
M
3
8.5
10
F
1
8.0
11
F
2
8.1
12
F
3
6.2
MEAN
8.0
SD
0.9
Graph No.2 : RBC Value (10
12
/L)
Table No. 3 – Haematocrit Values (%) :
HCT
Group
Animal No
Sex
Marking
%
NC
1
M
1
64.6
2
M
2
70.5
3
M
3
67.0
4
F
1
64.7
5
F
2
68.1
6
F
3
59.0
MEAN
65.7
SD
3.9
DC
7
M
1
69.8
8
M
2
68.8
9
M
3
68.7
10
F
1
69.6
11
F
2
63.8
12
F
3
51.5
MEAN
65.4
SD
7.1
Graph No.3 : Haematocrit Values (%)
Table No. 4 – Platelet Values (10
9
/L) :
PLT
Group
Animal No
Sex
Marking
*10
9
/L
NC
1
M
1
1116.0
2
M
2
726.0
3
M
3
804.0
4
F
1
565.0
5
F
2
838.0
6
F
3
782.0
MEAN
805.2
SD
180.1
Graph No.1 : Hb% Values (G/Dl)
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PLT
Group
Animal No
Sex
Marking
*10
9
/L
DC
7
M
1
935.0
8
M
2
846.0
9
M
3
927.0
10
F
1
671.0
11
F
2
919.0
12
F
3
484.0
MEAN
797.0
SD
182.8
Table No. 5 – MID Value (10
9
/L) :
MID
Group
Animal No
Sex
Marking
*10
9
/L
NC
1
M
1
1.6
2
M
2
2.4
3
M
3
1.1
4
F
1
1.4
5
F
2
1.2
6
F
3
1.8
MEAN
1.6
SD
0.5
DC
7
M
1
3.0
8
M
2
1.3
9
M
3
2.7
10
F
1
0.5
11
F
2
0.8
12
F
3
1.1
MEAN
1.6
SD
1.0
Graph No.5 : MID Value (10
9
/L)
Table No. 6 – Total WBC Value (10
9
/L) :
WBC
Group
Animal No
Sex
Marking
*10
9
/L
NC
1
M
1
17.5
2
M
2
17.5
3
M
3
15.4
4
F
1
16.1
5
F
2
16.3
6
F
3
13.9
MEAN
16.1
SD
1.4
DC
7
M
1
31.9
8
M
2
18.2
9
M
3
21.3
10
F
1
6.4
11
F
2
10.5
12
F
3
11.2
MEAN
16.6
SD
9.3
Graph No.4 : Platelet Values (10
9
/L)
Graph No.6 : Total WBC Value (10
9
/L)
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Table No. 7 – DLC-Neutrophil Value (10
9
/L) :
NEUT
Group
Animal No
Sex
Marking
*10
9
/L
NC
1
M
1
4.8
2
M
2
6.3
3
M
3
4.8
4
F
1
5.3
5
F
2
6.2
6
F
3
4.8
MEAN
5.4
SD
0.7
DC
7
M
1
11.0
8
M
2
6.5
9
M
3
6.8
10
F
1
2.0
11
F
2
3.2
12
F
3
3.6
MEAN
5.5
SD
3.3
Graph No.7 : DLC-Neutrophil Value (10
9
/L)
Table No. 8– DLC- Lymphocyte Value (10
9
/L) :
LYM
Group
Animal No
Sex
Marking
*10
9
/L
NC
1
M
1
11.1
2
M
2
8.8
3
M
3
9.5
4
F
1
9.4
5
F
2
8.9
6
F
3
7.3
MEAN
9.2
SD
1.2
PLT
Group
Animal No
Sex
Marking
*10
9
/L
DC
7
M
1
17.9
8
M
2
10.4
9
M
3
11.8
10
F
1
3.9
11
F
2
6.5
12
F
3
6.5
MEAN
9.5
SD
5.0
Discussion :
The mean Hb% observed in normal control is 15.7
g/dl whereas in Disease control, it is 15.3 g/dl.
Statistically not much significant decrease in Hb%
is observed. This Hb% count is in the normal range.
The mean RBC count observed in normal control
is 7.9 10
12
/L whereas in Disease control, it is 8
10
12
/L. No much variation in the values of RBC
count was observed in NC and DC group. This
RBC count is in the normal range.
The mean Haematocrit / PCV observed in normal
control is 65.7% whereas in Disease control, it is
65.4 %. No much variation in the values of PCV
count was observed in NC and DC group.
The mean Platelet value observed in normal
control is 805.2 10
9
/L whereas in Disease control, it
is 797 10
9
/L. No much variation in the values of
Graph No.8 : DLC- Lymphocyte Value (10
9
/L)
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Platelet count was observed in NC and DC group.
The mean MID value observed in normal control is
1.6 10
9
/L whereas in Disease control, it is 1.6
10
9
/L. No variation in the values of Platelet count
was observed in NC and DC group.
The mean WBC value observed in normal control
is 16.1 10
9
/L whereas in Disease control, it is 16.6
10
9
/L. No variation in the values of WBC count
was observed in NC and DC group.
The mean Neutrophil count observed in normal
control is 5.4 10
9
/L whereas in Disease control, it is
5.5 10
9
/L. No variation in the values of Neutrophil
count was observed in NC and DC group.
The mean Lymphocyte count observed in normal
control is 9.5 10
9
/L whereas in Disease control, it is
9.5 10
9
/L. No variation in the values of
Lymphocyte count was observed in NC and DC
group.
Summary & Conclusion:
There is no much variation in the values of blood
indices were noticed in NC and DC group. Hence,
it can be concluded that Bhaya (fear) may not act as
an etiological factor for the alteration of
hematological parameters in rats.
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Chaukhamba Sanskrit Pratishthan, ansari road,
Daryaganj, New Delhi – 110 002, 2019, pg.6
Declaration :
Conflict of Interest : None
ISSN: 2584-2757
DOI : 1 0 . 5 2 8 1 / z e n o d o . 1 7358633
Dr.
Subhash Waghe
Inter. J.Digno. and Research
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