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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. Elevated

triglyceride and elevation of low-density Cholesterol are the biomarkers used to know the risk of ischemic

heart disease. Cardiac biochemical parameters like CPK-MB, SGOT are measured to know the effect of

stressors on cardiac health. And accordingly, they are evaluated in both normal and disease control rats.

Cardiac biopsy was also done at the end of the study for further evaluation. Significant changes were

observed in disease control group indicating that fear lead to cardiac discomfort in experimental rats.

Keywords : Anxiety, Bhaya, Chinta ,CVD, Heart Disease

P

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Publisher

ROGANIDAN VIKRUTIVIGYAN PG ASSOCIATION

FOR PATHOLOGY AND RADIODIGNOSIS

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: 1 0 . 5 2 8 1 / z e n o d o . 1 6 0 3 0 973

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 Anxiety As Aetiological factor for Development of Heart

Disease In Albino Rats WSR To Lipid Profile And Cardiac Biochemistry and

Biopsy

Dr.Subhash Waghe

1

, Dr. Vijay R. Potdar

2

1

Professor & HOD – Dept. of Rognidana & Vikrutivigyna, Sardar Patel Ayurvedic Medical College & Hospital,

Dongariya, Balaghat – 44 3318 (M.P.)

2

Associate Professor - Dept. of Rognidana & Vikrutivigyna, Govt. Ayurvedic College, Nagpur.

Cite this article as: - Dr.Subhash Waghe (2025) ; Critical Evaluation of Anxiety As Aetiological factor for Development of Heart

Disease In Albino Rats WSR To Lipid Profile And Cardiac Biochemistry and Biopsy;Inter.J.Dignostics and Research 2 (4) 29-39,

DOI : 1 0 . 5 2 8 1 / z e n o d o . 1 6 0 3 0 973

G

A

R

V

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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. 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. 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. 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. Cardiac biochemical parameters

like CPK-MB, SGOT are measured to know the

effect of stressors on cardiac health. Cardiac biopsy

was also done at the end of the study for further

evaluation.

Review Of Literature :

Circulatory System As Per Ayurveda :

As per Ayurveda the root of Rasavaha strotas

(circulatory system) is mentioned as Heart and

blood vessels.

[3, 4]

As per acharya Charaka, Vyan

vayu circulates the blood in the entire body and it

gets aggravated whenever there is obstruction to the

flow of the blood in the circulation.

[5]

Aetiology of Ischemic Heart Diseases As Per

Ayurveda : As per acharya Charaka, psychological

factors like excessive worry along with

hyperlipidemic diet leads to the vitiation of

circulatory system

[6]

As per acharya Charaka,

excessive worries, fear, mental stress, chronic

disease leads and trauma leads to heart diseases.

[7]

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31

As per acharya Sushruta, excessive consumption of

incompatible diet, excessive diet, antagonistic diet

leads to heart diseases.

[8]

As per the book

Yogaratnakar, the smoking of tobacco leads to

heart diseases.

[9]

Atherosclerosis In Arteries As Per Ayurveda : In

Ayurveda, Dhamani -pratichaya (Atherosclerosis)

is defined as the excessive deposition of layer of

fatty sticky unctuous material inside the lumen of

arteries and it is the disease of Kapha origin. As

per Ayurveda, Dhamni Pratichay is one of the

diseases, caused exclusively by the vitiation of

Kapha (Kaphaj Nanatamaj Vyadhi)

[10, 11, 12]

Hence,

the factors, responsible for the vitiation of Kapha,

also serves as the aetiological factors for the

atherosclerosis in arteries (Dhamni Pratichaya). As

per acharya Charaka, it is Raspradoshaj Vikara and

it is due over nourishment.

[13, 14]

The function of

pathologically increased Kapha is to cause coating,

obstruction and hardness in the arterial lumen.

[15]

Aetiology of Ischemic Heart Diseases As Per

Ayurveda :

As per acharya Sushruta, due to consumption of

high fatty and carbohydrate diet and lack of

exercise, the arterial lumen gets obstructed with fat

and area to be supplied, remain under perfused.

[16]

As per acharya Sushruta, the vitiated plasma gets

obstructed due to blockages in coronaries of the

heart, and alters the normal functioning of the heart

and also gives rise to Angina.

[ 17]

The angina if not

treated soon, kills the patient instantly.

[18]

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.

[19] [20]

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]

CPK -MB is creatine phospho kinase myocardial

bound enzyme primarily found in heart muscle. It is

used to detect the heart muscle damage in

conditions such as myocardial infarction. It is

released in blood circulation when heart muscle is

damaged.Serum Glutamic Oxaloacetic Transferase

is an enzyme found in the liver and heart tissue. It’s

rise in blood indicates either liver or heart muscle

injury.Inflammatory biomarker C-Reactive is a

protein produced in the liver in response to the

inflammation and it is used to assess the presence

of inflammation in the body. HS-CRP is more

specific to the cardiac tissue injury.Triglyceride is

the stored form of fat which is used to derive

energy. Elevated triglyceride level indicates an

increased risk for ischemic heart disease. High

density lipoprotein is a good cholesterol which

helps to prevent the building of bad low density

cholesterol in the circulation. HDL picks up the

excess LDL cholesterol and send it to liver where it

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32

is broken down and eliminated. Low levels of

HDL cholesterol indicated increased risk for

ischemic heart disease.

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

development of heart disease

 Alternate Hypothesis (H0) : Ayurvedokta

Bhaya (fear) 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.

 Other Objectives :To study the

aetiopathogenesis of myocardial infarction

from Ayurvedic point of view.

Material & Methodology :

7.1 Study Design

Center of Study – Dept of Roga Nidana &

Vikrutvigyana, Government Ayurvedic College,

Nanded And National Testing Centre, Pune

Duration of Study – Total study 18 months after

approval of synopsis.

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 are

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 are randomly scheduled over a one

week period and are 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. The normal reference range of CPK-MB in

rats is 5 to 25 IU/L. Level. The level 5 times

higher than the normal range is considered

significant rise.

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2. The normal reference range of SGOT in rats

is 5 to 40 IU/L. Level. The level higher than

the normal range will be considered

significant rise.

3. The normal reference range of CRP in rats

is 300 to 600 mg/ml. Level. The level

higher than the normal range is considered

significant rise.

4. The normal reference range of Triglyceride

in rats is 25 to 145 mg/dl. Level. The level

higher than the normal range is considered

significant rise.

5. The normal reference range of HDL

cholesterol in rats is 35 to 55 mg/dl. Level.

The level higher than the normal range is

considered significant rise.

[21]

Observation & Result:

Table No. 1 – CPK-MB Values

Date:07/Feb/2024

CKMB

Group

Animal No

Sex

Marking

U/L

NC

1

M

1

429.8

2

M

2

373.0

3

M

3

410.0

4

F

1

488.7

5

F

2

421.4

6

F

3

370.1

MEAN

415.5

SD

43.6

DC

7

M

1

665.0

8

M

2

706.7

9

M

3

645.1

10

F

1

651.4

11

F

2

710.8

12

F

3

845.8

MEAN

704.1

SD

74.7

Graph 1: CPK-MB Values

Table No. 2 – CRP Values

Date:07/Feb/2024

CRP

Group

Animal No

Sex

Marking

mg/L

NC

1

M

1

0.7

2

M

2

0.9

3

M

3

0.4

4

F

1

0.7

5

F

2

0.4

6

F

3

0.8

MEAN

0.6

SD

0.2

DC

7

M

1

1.6

8

M

2

1.7

9

M

3

1.9

10

F

1

1.9

11

F

2

1.6

12

F

3

1.8

MEAN

1.8

SD

0.1

Graph 2: CRP Values

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Table No. 3 – SGOT Values

Date:07/Feb/2024

SGOT

Group

Animal No

Sex

Marking

U/L

NC

1

M

1

54.0

2

M

2

55.1

3

M

3

52.4

4

F

1

48.6

5

F

2

51.5

6

F

3

65.4

MEAN

54.5

SD

5.8

DC

7

M

1

104.8

8

M

2

172.2

9

M

3

123.3

10

F

1

104.4

11

F

2

112.4

12

F

3

128.6

MEAN

124.3

SD

25.4

Graph 3: SGOT Values

Table No. 4 – Triglyceride Values

Date:07/Feb/2024

TGL

Group

Animal No

Sex

Marking

mg/dl

NC

1

M

1

170.7

2

M

2

186.2

3

M

3

140.9

4

F

1

164.5

5

F

2

183.1

6

F

3

142.2

MEAN

164.6

SD

19.5

DC

7

M

1

182.0

8

M

2

206.5

9

M

3

157.8

10

F

1

239.3

11

F

2

242.5

12

F

3

214.1

MEAN

207.0

SD

32.9

Graph 4: Triglyceride Values

Table No. 5 – HDL Values

Date:07/Feb/2024

HDL

Group

Animal No

Sex

Marking

mg/dl

NC

1

M

1

34.2

2

M

2

28.5

3

M

3

32.1

4

F

1

70.4

5

F

2

61.1

6

F

3

57.8

MEAN

47.4

SD

17.8

DC

7

M

1

18.5

8

M

2

35.7

9

M

3

30.6

10

F

1

39.0

11

F

2

44.7

12

F

3

57.4

MEAN

37.7

SD

13.2

Graph 5: HDL Values

HDL

NC

DC

0

20

40

60

ns

Groups

HDL (mg/dl)

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Sr

.

N.

Group

/Slide

Code

Histopathological

observations

1.Heart

Over

all

Patho

logica

l

Grad

e

1

NC-

1M

Normal

histomorphological

features of cardiac

muscle fibers in

the myocardium.

Absence of

inflammatory or

pathological

changes in heart

tissue.

NAD

2

NC-2F

Normal

histomorphological

features of cardiac

muscle fibers in

the myocardium.

Absence of

inflammatory or

pathological

changes in heart

tissue.

NAD

3

DC_3

M

Mild degenerative

changes in the

cardiac muscle

fibers.

Multifocal areas of

congestion and

occasional foci of

interstitial

hemorrhages in

pericardium and

myocardium

region.

Mild

(+2)

4

DC-4F

Mild degenerative

changes in the

cardiac muscle

fibers with focal

areas of congestion

and occasional foci

of interstitial

hemorrhages in

pericardium and

myocardium

region.

Mini

mal

(+1)

to

Mild

(+2)

Image 1: Cardiac Biopsy – NCM

Image 2: Cardiac Biopsy – DCM

Discussion :

The mean CPK-MB observed in normal control is

415.5 U/L whereas in Disease control, it is 704.1

U/L. Statistically significant increase in CPK-MB

is observed. CPK -MB is creatine phospho kinase

myocardial bound enzyme primarily found in heart

muscle. It is used to detect the heart muscle damage

in conditions such as myocardial infarction. It is

released in blood circulation when heart muscle is

damaged. The normal reference range of CPK-MB

in rats is 5 to 25 IU/L. Level. The level 5 times

higher than the normal range will be considered

significant rise.The mean CRP observed in normal

control is 0.6 mg/L whereas in Disease control, it is

1.8. mg/L. Statistically significant increase in

inflammatory biomarker C-Reactive Protein is

observed. CRP is a protein produced in the liver in

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response t the inflammation and it is used to assess

the presence of inflammation in the body. HS-CRP

is more specific to the cardiac tissue injury. The

normal reference range of CRP in rats is 300 to 600

mg/ml. Level. The level higher than the normal

range will be considered significant rise.

The mean SGOT observed in normal control is 54.5

U/L whereas in Disease control, it is 124.3 U/L.

Statistically significant increase in SGOT is

observed. Serum Glutamic Oxaloacetic Transferase

is an enzyme found in the liver and heart tissue. It’s

rise in blood indicates either liver or heart muscle

injury. The normal reference range of SGOT in rats

is 5 to 40 IU/L. Level. The level higher than the

normal range will be considered significant rise.

The mean triglyceride observed in normal control is

164.6 mg/dl whereas in Disease control, it is 207.0

mg/dl. Statistically significant increase in

triglyceride is observed. Triglyceride is the stored

form of fat which is used to derive energy. Elevated

triglyceride level indicate an increased risk for

ischemic heart disease. The normal reference range

of Triglyceride in rats is 25 to 145 mg/dl. Level.

The level higher than the normal range will be

considered significant rise.The mean high density

cholesterol observed in normal control is 47.4

mg/dl whereas in Disease control, it is 37.7 mg/dl.

Statistically significant decrease in protective high

density cholesterol is observed. High density

lipoprotein is a good cholesterol which helps to

prevent the building of bad low density cholesterol

in the circulation. HDL picks up the excess LDL

cholesterol and send it to liver where it is broken

down and eliminated. Low levels of HDL

cholesterol indicated increased risk for ischemic

heart disease. The normal reference range of HDL

cholesterol in rats is 35 to 55 mg/dl. Level. The

level higher than the normal range will be

considered significant rise.In cardiac biopsy, mild

degenerative changes in the cardiac muscle fibers

with focal areas of congestion and occasional foci

of interstitial hemorrhages in pericardium and

myocardium region were seen. This again suggests

the cardiac damage induced by the stressors.

Summary & Conclusion :

1. In the DC group, levels of CK-MB, CRP,

SGPT, SGOT, Triglycerides were significantly

elevated as compared to the NC group.

2. No significant difference was observed in HDL,

Total ides (Cl) in the DC group as compared to

NC.

3. Histopathology of heart tissue showed no

abnormalities in NC group. While in DC group

Mild (+2) degenerative changes in the cardiac

muscle fibers. Multifocal areas of congestion

and occasional foci of interstitial hemorrhages in

pericardium and myocardium region were

observed

4. On the basis of the ECG parameters like RR

interval and QT interval results obtained, it can

be concluded that fear can acts as an etiological

factor for the development of heart disease in

rats.

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Issue : 04

INTERNATIONAL JOURNAL OF DIAGNOSTICS AND RESEARCH [ISSN No.: 2584-2757]

Volume : 02

Copyright @ : - Dr.Subhash Waghe Inter. J.Digno. and Research IJDRMSID00047 |ISSN :2584-2757

39

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