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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. ECG is the commonest diagnostic tool
which is used to diagnose cardiac abnormalities in both humans and animals. In present study, ECG was
done using ketamine anaesthesia and all measurements are decided accordingly. After confirmation of stress
in animals, Electrocardiography parameters are checked using Data Acquisition System, Powerlab.
Significant changes were observed in disease control group indicating that fear lead to cardiac discomfort in
experimental rats.
Keywords : Chinta , Bhaya ,Manasik Trass, ECG
P
ISSN No. : 2584-2757
Volume : 02
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DOI
: 1 0 . 5 2 8 1 / z e n o d o . 16031259
Reg. No. : MAHA-703/16(NAG)
Year of Establishment – 2016
INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH
Corresponding author: Dr.Subhash Waghe
Article Info: Published on : 15/07/2025
Impact Factor : 1.013
Critical Evaluation of Fear As Aetiological factor for Development of Heart
Disease In Albino Rats WSR To Electrocardiography
Dr.Subhash Waghe
1
1
Professor & HOD – Dept. of Rognidana & Vikrutivigyna, Sardar Patel Ayurvedic Medical College & Hospital,
Dongariya, Balaghat – 44 3318 (M.P.)
Cite this article as: - Dr.Subhash Waghe (2025) Critical Evaluation of Fear As Aetiological factor for Development of Heart
Disease In Albino Rats WSR To Electrocardiography ;Inter .J .Dignostics and Research 2 (4) 40- 48, DOI
:1 0 . 5 2 8 1 / z e n o d o . 16031259
G
A
R
V
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41
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]
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 and Syndrome of MI is
mentioned by Sushrut Uttartantra 43/131-132 in the
form of ‘Hrit Shoola’. 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.
[2, 3]
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
(Fear/Anxiety) in the development of heart disease.
Stress is an important factor having high impact on
the psychological development which alters emotion,
cognition and related behavioral outputs.
The Chronic Unpredictable Mild Stress (CUMS)
model in rats is a widely used animal model for
inducing depressive-like behaviors by exposing
rodents to a series of random, mild stressors over
several weeks. This model is designed to mimic the
cumulative effects of daily life stressors that
contribute to anxiety and depression in humans.
CUMS leads to disruption of homeostasis, causing
somatic, physiological, neurobiological,
biochemical, and behavioral disturbances.
[4]
ECG
is the most popular method of knowing the heart
rhythm and ischemic abnormalities. In rats, ECG is
recorded with or without giving the anaesthesia to
the rats. Telemetry is the method which records the
ECG without giving the anaesthesia. in present
study, ECG was done using ketamine anaesthesia
and all measurements are decided accordingly. After
confirmation of stress in animals,
Electrocardiography parameters are checked using
Data Acquisition System, Power lab.
Review Of Literature :
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. 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 is 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.
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42
These stressors are randomly scheduled over a week
period and are repeated to maintain the aspect of
unpredictability. At the end of every week sucrose
consumption test and body weight of all animal are
measured to confirm the induction of stress in
animals.
[ 5]
There are several invasive and non invasive
techniques to record 1 to 12 channel ECG recordings
in rats. Most studies use limb lead II which is
sufficient for general analysis of ECG parameters in
rats. Surface ECG recordings is the most commonly
used technique in anesthetized rats. To obtain a limb
lead ECG, the electrodes are placed under the skin of
left and right forepaws and the tail. In this technique,
measurements may be confounded by type of
anaesthesia. Telemetry is the method which records
the ECG without giving the anaesthesia. Telemetry
transmitters are implanted subcutaneously in the
abdominal cavity or intrascapular region whereas
electrodes connected to the transmitters are placed
the anterior mediastinum. Data from transmitters are
gathered wirelessly by a receiver placed outside the
rat cage. This method provides data that is free of
aneasthesia.
[6]
However, in present study, ECG was
done using ketamine anaesthesia and all
measurements are decided accordingly. After
confirmation of stress in animals,
Electrocardiography parameters are checked using
Data Acquisition System, Power lab.
Myocardial Infarction (MI) :
MI refers to the condition where there is imbalance
between the myocardial oxygen demand and its
supply due to the obstruction of blood supply in
coronary arteries.
[7] [8]
The commonest causes responsible for it are :
Atherosclerosis in coronary artery
Thrombosis
Investigations To Diagnose MI :
Lipid profile – It may show dyslipidaemia
(Increased LDL cholesterol and
Triglycerides)
Cardiac Markers - Serum Troponin and
CPK-MB elevated.
ECG shows ST-T changes. In rats ST
segment is absent in waveforms.
Coronary Angiography (CAG) shows
coronary occlusions.
2-D Echocardiography shows regional wall
motion abnormalities.
[20 & 21]
Research Question :
Whether Ayurvedokta Bhaya (fear) acts as a
aetiological factor for development of heart disease.
Hypothesis :
Null Hypothesis (H1):
Ayurvedokta Bhaya (fear) factor acts as a
aetiological factor for development of heart
disease
Alternate Hypothesis (H0):
Ayurvedokta Bhaya (fear) factor does not
acts as an aetiological factor for development
of heart disease.
Aims & Objectives :
Primary Objectives:
The present study, aims to study the
aetiological factor Bhaya (Fear/Anxiety) as
the cause for the development of heart
disease.
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43
Other Objectives:
To study the aetiopathogenesis of myocardial
infarction from Ayurvedic point of view.
Material & Methodology :
Study Design :
Center of Study – Dept of Roga Nidana &
Vikrutvigyana, Government Ayurvedic College,
Nanded And National Testing Centre, Pune
Duration of Study – 18 months
Study Population And Sampling
Animal required for the Study
Species/Common name - Albino Rat
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, ECG was done using the power
Lab data acquisition apparatus on 0, 28
th
and 49
th
Day. Rats were anaesthetized with Ketamine before
taking the ECG.
Assessment Criteria:
1. ECG is monitored in 8 Channel power laboratory
(Data Acquisition system) and different ECG
parameters are measured.
RR interval is the time between the two consecutive
R wave peaks. In rats Heart rate is calculated using
RR interval only because rat ECG lacks the Q
waves. (HR = 60/ (R-R interval in seconds). In
matured rats RR interval is 118-251ms.
PR interval is measured from the beginning of the P
wave to the beginning of the QRS/RS complex. The
PR interval in rats ranges from 38 to 70 ms. It is
significantly affected by the type of anaesthesia
used. It is 56 to 66 ms in rats anaesthetized with
Ketamine.
QRS complex is located between Q and S wave. It
represents the time taken by wave of depolarization
to move through the ventricles. Narrowing of QRS is
seen in supraventricular arrhythmias whereas
widening of QRS is seen in Ventricular arrhythmia
and bundle branch blocks. Since Q wave is not
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usually detectable in Rat ECG, usually the RS
complexes are evaluated. The duration of QRS in
rats under ether anesthesia is 11.3to 16.1 ms. And 12
to 15.7 in rats anaesthetized with Ketamine.
QT interval is the time taken from the Q wave to the
end of the T wave. In rats this parameter is usually
measured from the onset of Rs complex to the end of
T wave. QT interval represents the time of
depolarization and repolarization of ventricles. It
may be affected due to intrinsic heart diseases or
drug toxicity. A prolonged QT interval in rats also
been found in hypokalemia and myocardial
infarction. QT interval in rats is also affected by the
type of anaesthesia used. In SD rats it is 50-70ms.
Whereas in rats anesthetized with ketamin, it is
found to be 75-95ms. In rats anesthetized with ether,
it is found to be 60.6- 62.5 ms.
Observation & Result :
Table No. 1 – RR Interval (Sec) :
Days
Day
0
Day
28
Day
49
Group
Ani.
No
RR
RR
RR
NC
1
0.222
0.217
0.225
2
0.254
0.257
0.252
3
0.233
0.239
0.235
4
0.259
0.254
0.265
5
0.237
0.248
0.246
6
0.249
0.251
0.255
Mean
0.242
0.244
0.246
SD
0.014
0.015
0.014
DC
7
0.251
0.253
0.258
8
0.235
0.232
0.228
9
0.234
0.230
0.218
10
0.248
0.247
0.221
11
0.252
0.243
0.231
12
0.250
0.249
0.232
Mean
0.245
0.242
0.231
SD
0.008
0.009
0.014
Graph 1: RR Interval (Sec)
Table No. 2 – PR Interval (Sec) :
Days
Day
0
Day
28
Day
49
Group
Ani.
No
PR
PR
PR
NC
1
0.049
0.050
0.059
2
0.052
0.053
0.022
3
0.048
0.051
0.048
4
0.050
0.050
0.045
5
0.049
0.013
0.051
6
0.050
0.054
0.052
Mean
0.050
0.045
0.046
SD
0.001
0.016
0.013
DC
7
0.056
0.044
0.040
8
0.055
0.052
0.047
9
0.056
0.046
0.039
10
0.051
0.050
0.048
11
0.045
0.043
0.042
12
0.049
0.048
0.042
Mean
0.052
0.047
0.043
SD
0.004
0.004
0.004
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Graph 2: PR Interval (Sec)
Table No. 3 – QRS (Sec) :
Days
Day
0
Day
28
Day
49
Group
Ani.
No
QRS
QRS
QRS
NC
1
0.026
0.019
0.015
2
0.014
0.029
0.035
3
0.026
0.007
0.024
4
0.025
0.023
0.022
5
0.016
0.023
0.036
6
0.020
0.038
0.014
Mean
0.021
0.023
0.024
SD
0.005
0.010
0.009
DC
7
0.023
0.022
0.025
8
0.022
0.023
0.026
9
0.012
0.023
0.023
10
0.025
0.026
0.027
11
0.023
0.028
0.032
12
0.020
0.032
0.039
Mean
0.021
0.026
0.029
SD
0.005
0.004
0.006
Table No. 4 – QT Interval (Sec) :
Days
Day
0
Day
28
Day
49
Group
Ani.
No
QT
QT
QT
NC
1
0.021
0.021
0.021
2
0.016
0.018
0.020
3
0.016
0.018
0.016
4
0.025
0.022
0.018
5
0.020
0.016
0.019
6
0.016
0.018
0.015
Mean
0.019
0.019
0.018
SD
0.004
0.002
0.002
DC
7
0.021
0.015
0.014
8
0.021
0.016
0.015
9
0.021
0.018
0.016
10
0.020
0.018
0.012
11
0.014
0.017
0.013
12
0.020
0.020
0.018
Mean
0.020
0.017
0.015
SD
0.003
0.002
0.002
Graph 3: QRS (Sec) :
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Discussion :
Long-term exposure to stressful conditions is
associated with the development of a manifold of
pathophysiological conditions, including those
affecting behaviour, immune physiology, neuronal
signalling, and cardiovascular function as well as
chronic mood disorders such as anxiety and
depression. (Glaser & Kiecolt-Glaser, 2005)
[9]
This requires animal models to validate the casualty
between stress and overt development of heart
disease. 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. Accordingly, this model was
used in present study.
[10]
Animals were subjected 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 are repeated to maintain the aspect of
unpredictability.
As per researches, CUMS model leads to many
behavioral changes and changes in brain structure
and function, including alterations in
neurotransmitter systems (e.g., serotonin, dopamine,
and norepinephrine), neurotrophic factors, and stress
hormone levels. The unpredictable nature of the
stressors more closely resembles the chronic,
unpredictable stressors experienced in real human
life. ECG is the most popular method of knowing
the heart rhythm and ischemic abnormalities.
In rats, ECG is recorded with or without giving the
anaesthesia to the rats. In present study, ECG was
done using ketamine anaesthesia and all
measurements are decided accordingly.
The mean RR interval observed in normal control on
day 0 is 0.242 whereas in Disease control, it is 0.245.
The mean RR interval observed in normal control on
day 14 is 0.244 whereas in Disease control, it is
0.242.
The mean RR interval observed in normal control
on day 42 is 0.246 whereas in Disease control, it is
0.231. RR interval found to be decreased on day 28
in DC which was not statistically significant. While
on Day 49 statistically significant decrease was seen
in DC as compared to NC indicating the effect of
stressors on RR Interval.
The mean PR interval observed in normal control on
day 0 is 0.050 whereas in Disease control, it is 0.052.
The mean PR interval observed in normal control on
day 14 is 0.045 whereas in Disease control, it is
0.047. The mean PR interval observed in normal
control on day 42 is 0.046 whereas in Disease
control, it is 0.043. On all the three measurements
there is rise in blood pressure in disease control
indicating that stress had increased the blood
pressure. The mean QRS observed in normal control
on day 0 is 0.021 whereas in Disease
Graph 4: QT Interval (Sec)
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control, it is 0.021. The mean QRS observed in
normal control on day 14 is 0.023 whereas in Disease
control, it is 0.026. The mean QRS observed in
normal control on day 42 is 0.024 whereas in Disease
control, it is 0.029.
No significant change in the PR & QRS interval is
observed.
The mean QT interval observed in normal control on
day 0 is 0.019 whereas in Disease control, it is 0.020.
The mean QT interval observed in normal control on
day 14 is 0.018 whereas in Disease control, it is
0.017. The mean QT interval observed in normal
control on day 42 is 0.018 whereas in Disease
control, it is 0.015.
QT interval has decreased on day 28 but not
statistically significant.
Summary & Conclusion :
1. RR interval found to be decreased on day 28 in
DC which was not statistically significant. While
on Day 49 statistically significant decrease was
seen in DC as compared to NC indicating the
effect of stressors on RR Interval.
2. PR- No significant change in the PR & QRS
interval. These results indicate no change in
stress.
3. QT interval has decreased on day 28 but not
statistically significant.
4. On the basis of the ECG parameters like RR
interval and QT interval results obtained, it can be
concluded that Bhaya fear can acts as an
etiological factor for the development of heart
disease in rats.
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