The Aggregation response of Platelets in different...

AAMJ / Vol. 3 / Issue 6 / November – December 2017

A A M J Anveshana Ayurveda Medical Journal

www.aamj.in ISSN: 2395-4159

Research Article

The Aggregation response of Platelets in different Prakṟti

Aditi Yashwant Pimpalkhare 1 Ravindra Pandurang Bhurke 2 Lomesh Pitambar Gedam 3

A b s t r a c t

Background: Ayurveda believes that the path to optimal health is different for each person

depending upon their constitution or Prakṟti. On the basis of Prakṟti, the population can be

categorized into several subgroups. The concept is claimed to be useful in predicting an indi-

vidual’s susceptibility to a particular disease, prognosis and selection of therapy. The present

study was carried out to study if the platelet Aggregation response varies in different prakṟti

subtypes. Methods: After obtaining Institutional Ethics Committee permission, normal healthy

individuals of either sex between the age group of 16 to 40 years were recruited in the study.

Their prakṟti evaluation was done using a standardized validated questionnaire as per the soft-

ware AYUSOFT, developed by CDAC, Pune. Their Platelet Rich Plasma (PRP) was incubated.

Platelets in the PRP were counted using a platelet counter. This working plasma was used to

study the Aggregation response of platelets to Adenosine Diphosphate in concentration of

5µM/L & 10 µM/L. Platelet aggregation was studied using Chronolog platelet aggregometer.

Results: We observed that in the study population of normal healthy participants (n= 90), ADP-

induced platelet aggregation was highest among the Kapha prakṟti individuals as compared to

the other prakṟti types. From statistical calculation using One-way Analysis of Variance

(ANOVA), we conclude that, difference between platelet aggregation of three different prakṟti

is extremely significant (P<0.0001) at 5 µM & (P<0.0002) at 10 µM concentration of ADP.

Conclusion: Our results suggest that identifying the prakṟti may help in individualizing therapy

or predicting proneness to a disease.

Keywords: Prakṟti, Platelet aggregation, Adenosine diphosphate, AYUSOFT, Chronolog platelet

aggregometer

1 & 3 Assistant Professor at D. Y. Patil University, School of Ayurveda, Nerul, Navi Mumbai, 2 Assistant Professor at R. A. Podar Medical College & Hospital, Worli, Mumbai, India.

CORRESPONDING AUTHOR

Dr. Aditi Yashwant Pimpalkhare

Assistant Professor,

D. Y. Patil University, School of Ayurveda,

Nerul,

Navi Mumbai, (India).

Email: [email protected]

http://aamj.in/wp-content/uploads/Volume3/Issue6/AAMJ_1600_1604.pdf

Pimpalkhare et.al,. : The Aggregation response of Platelets in different Prakṟti

AAMJ / Vol. 3 / Issue 6 / November – December 2017 1601

INTRODUCTION

yurveda is a science of self-understanding. Ayur-

vedic approach is very individualized, since the

path to optimal health is different for each person

depending upon their unique constitution or Prakṟti. Ayur-

veda believes that the individual’s constitution (Prakṟti) is

framed at the time of conception as a genetic code or

predominant Panchamahabhautic doṣa that can be ex-

pressed physically and mentally as disease proneness

and emotional response. (I) To achieve our aim of life it is

necessary that our body and mind remains healthy. For

that we need to know our Prakṟti because Prakṟti influ-

ences every aspect of all living beings from birth to death.

So a fine understanding of principles behind Prakṟti is

unavoidable.

Haemostatic function of blood is a unique feature as it is

the first stage of wound healing. This involves coagula-

tion, blood changing from a liquid to a gel. It involves

three stages mainly: 1. Vasoconstriction 2. Platelet ag-

gregation to form a plug 3. Clot formation. Platelet ag-

gregation is the temporary stoppage of bleeding; clot is

the permanent one. But we see time duration for stop-

page of bleeding differs from person to person. Aim of

this study is to find the relation between Prakṟti & platelet

aggregation. The major component involved in platelet

aggregation is ADP (Adenosine Diphosphate). So ADP

induced Platelet aggregation is calculated for each indi-

vidual of known prakṟti.

Thrombosis is a pathological extension of normal haemo-

stasis. Haemostasis is the arrest of haemorrhage in re-

sponse to a vascular injury. Thrombosis is a formation of

a blood clot in circulation. More is the platelet aggrega-

tion; more is the chance of Atherosclerosis. Less is the

platelet aggregation, more is the chance of haemor-

rhage. Using this study, we can identify which Prakṟti is

more prone to Atherosclerosis or bleeding disorders and

advise them to prevent further consequences with the

help of Āhāra, Vihāra, profession and drugs to avoid

complications. Also useful in explaining the variable out-

come of the antiplatelet drug therapy.

METHODS

Sample Size:

Total number of volunteers included in the study were 90.

Appropriate number of volunteers were screened so as

to recruit 30 volunteers of each Ekadoṣapradhan prakṟti

type by simple random sampling.

Study Population:

Normal healthy individuals (confirmed by history, physi-

cal examination and routine laboratory investigations, in-

cluding haematology, renal and liver function tests) be-

tween the age group of 16 to 40 years (both years inclu-

sive) of either sex, and willing to abide by trial proce-

dures were enrolled.

Plan of Study:

After taking the consent, Prakṟti was evaluated using

standardized and validated software AYUSOFT, devel-

oped by CDAC, Pune. Software gives the result as Per-

centage of each doṣa calculated. Individuals having

60% or more characters included in the sample as doṣa

pradhan prakṟti of that doṣa.

Then blood collection was done in fasting condition to

minimize circadian variation in platelet aggregability.

Blood samples were collected between 8 and 9 am from

an antecubital vein in the supine position. 20cc blood

collection was done from each volunteer; 10 ml in Citrate

bulb for Platelet aggregation; 5 ml in Plain bulb for Bio-

chemical examination; 5 ml in EDTA bulb for Hematol-

ogy. Routine haematological and biochemical investiga-

tions were done for confirmation of their healthy status.

Their Platelet Rich Plasma was incubated. Platelets in the

PRP were counted using a platelet counter. This working

plasma was used to study the Aggregation response of

platelets to Adenosine Diphosphate in concentration of

5M & 10M.

Image 1: Chronolog platelet aggregometer

Platelet aggregation was studied using Chronolog plate-

let aggregometer which works on the principle of Born

GVR turbidimetric method. (II) Aggregometer measures

platelet function on blood samples using electrical imped-

ance in whole blood or optical density in Platelet Rich

Plasma (PRP) with simultaneously measuring ATP release

by the luminescence method. The output can be con-

nected to either a strip chart recorder or to a computer.

This program performs computation of the traces report-

ing the results as maximum amplitude and slope. Lag time

and area under the curve analysis is also available for

each trace. Each graph, its values, patient demographics

and reagent information are then stored on computer

disk for later retrieval and printing. The front panel has a

A



24-character Liquid Crystal Display (LCD) for each chan-

nel. Aim is to study the response of platelets to Adenosine

diphosphate (ADP). The method was standardised with

two concentrations of ADP i.e. 5µMol/L and 10µMol/L.

OBSERVATIONS & RESULTS

A total of 90 participants were recruited in the study. The

distribution of these volunteers according to Prakṟti along

with their demographic data is shown in Table 1. Inter-

estingly, the BMI in vāta prakṟti was significantly less as

compared to kapha & pitta prakṟti. In Pitta Prakṟti indi-

viduals’ platelet count was less & bleeding time was

more. BMI was high in Kapha Pradhan prakṟti.

Table 1: Demographic & Haematological details

Prakṟti n (M:F)

Platelet

count

lacks

/mm3

Bleeding

time mins BMI

Vātapra-

dhan

30

(16:14)

2.76 ±

0.59 1.7 ± 0.5

20.21

±

2.87

Pittapra-

dhan 30(13:17)

2.67 ±

0.64 2 ± 0.5

20.91

±

2.69

Kaphapra-

dhan

30

(11:19)

2.78 ±

0.70 1.9 ± 0.4

22.10

±

3.55

Prakṟti Wise Average Platelet Aggregation:

Difference in the Average Platelet aggregation was sig-

nificant at 5 µM ADP concentration than at 10 µM con-

centration.

Table 2: Prakṟti wise Average Platelet Aggregation

Prakṟti

Average Platelet aggregation % ±

S. D.

5 µM 10 µM

Vātapradhan 63% ± 14 75% ± 10

Pittapradhan 52% ± 18 70% ± 13

Kaphapra-

dhan 72% ± 6.8 81% ± 7.5

Statistical analysis of average platelet aggregation in dif-

ferent Prakṟti:

H0 = Platelet aggregation is independent on Doṣapra-

dhan Prakṟti

H1 = Platelet aggregation is dependent on Doṣapradhan

Prakṟti

One-way Analysis of Variance ANOVA test is applied to

determine the Significance in Doṣapradhan Prakṟti in IN-

STAT software. The level of significance is set at 5% level,

P<0.05 level. Fcalculated > Ftable value indicates the signifi-

cance and it reveals that the effect is due to Doṣapra-

dhana Prakṟti.

At ADP Concentration 5 µM:

The P value is < 0.0001, considered extremely signifi-

cant. Variation among column means is significantly

greater than expected by chance.

ANOVA table

Source of var-

iation

Degrees of

freedom

Sum of

squares

Mean

square

between col-

umns 2 6620 3310

within columns 87 16834 193.5

Total 89 23454

F = 17.106

Table value for 2 df across and 87 df vertically at 5%

level of significance =3.101. Calculated value is greater

than table value. So null hypothesis H0 is rejected and

alternate hypothesis of significant difference between the

mean is accepted hence Doṣapradhan Prakṟti and Plate-

let aggregation are dependent and there is significant

difference in the Platelet aggregation in Vāta, Pitta and

Kapha pradhan Prakṟti at 5µm ADP concentration.

At ADP Concentration 10µM:

The P value is 0.0002, considered extremely significant.

Variation among column means is significantly greater

than expected by chance.

ANOVA table

Source of

variation

Degrees of

freedom

Sum of

squares

Mean

square

between col-

umns 2 2180 1090

within col-

umns 87 9389 107.92

Total 89 11569

F = 10.100

Table value for 2 df across and 87 df vertically at 5%

level of significance =3.101. Calculated value is greater

than table value. So null hypothesis H0 is rejected and

H1 of significant difference between the mean is ac-

cepted hence Doṣapradhan Prakṟti and Platelet aggrega-

tion are dependent and there is significant difference in

the Platelet aggregation in Vāta, Pitta and Kapha pra-

dhan Prakṟti at 10µm ADP concentration.



Graph 1: Platelet count versus Prakṟti

Image 2: Platelet aggregation of Vātaprakṟti:

Image 3: Platelet aggregation of Kaphaprakṟti:

Image 4: Platelet aggregation of Pittaprakṟti:

DISCUSSION

Everybody imparts different response to the internal & ex-

ternal disease causing agents. This response is depend-

ent on Prakṟti, the basic constitution of the body. These

prakṟtis show characteristics of the dominant Doṣa at an-

atomical, physiological and psychological levels. For the

transport purpose of blood, it has to be kept in liquid

form. This Dravata is the Aapmahabhutjanya guna of

Raktadhātu. But its degree will differ in each person ac-

cording to prakṟti. Pitta & Kapha are the two Doṣas

which are in liquid form circulate in the body with the

medium of blood. Vitiation of these two doṣas directly

affects the Dravata of Raktadhātu. (III) So in Pitta prakṟti

individuals, Raktadhātu will be more affected by Pitta

doṣa rather than Kapha doṣa. Atherosclerotic changes

(plaques) in the vessel or excessive platelet aggregation

will affect the Dravata of Raktadhātu. In that Kapha will

cause Atherosclerotic plaques similar to its guna & pitta

will cause internal bleeding or emboli formation. Since

Pitta has the “Aashrayashrayisambandh” with Rak-

tadhātu. So whenever Pitta is vitiated, it affects Rakta

dhātu and vice versa. (IV)

Sushruta says normal blood does not coagulate inside

the blood vessel. (V)(VII) It is always in liquid form. (IX) So

from this we can say consistency of blood is dependent

on healthy state of your body. According to Sushruta, in

all individuals with any Prakṟti, Raktadhātu is Asamhata.

It is neither a clear fluid nor dense. But the degree of

consistency may differ in different individuals. It is Prakṟti

dependent. (VI)

Here Sushruta clearly says that normal blood can prevent

bleeding. This we can correlate with the coagulation pro-

cess. But its degree will minutely differ according to

Prakṟti. We have observations that platelet aggregation

is maximum in Kapha Prakṟti & minimum in Pitta prakṟti

individuals. As Charaka has said Pitta dushta Rakta

shows delay in clotting. This can be seen as less platelet

aggregation in Pitta Prakṟti individuals. And Kapha

dushta Rakta is thick (VIII) or does not flow easily. (X) This

can be correlated with more platelet aggregation in

Kapha Prakṟti individuals.

While explaining bloodletting procedure, it is given that

vāta & pitta vitiated rakta is slow in aggregation or have

properties that prevent aggregation. (Dry, fast moving,

does not clot easily) (VI) (IX) In contrast, Kapha vitiated rakta

has properties in favour of aggregation of platelets.

(Cold, slimy, bleeding time is more, slow moving) (VI) So

from this study we can conclude that, Kapha prakṟti sub-

jects have maximum platelet aggregation. And pitta

prakṟti subjects have the least platelet aggregation.

1.5

2

2.5

3

3.5

VATAPITTA

KAPHA

2.762.67 2.78

Pla

tele

t co

un

t in

la

cks/

dl

Prakriti

PLATELET COUNT



Platelet aggregation varies between 18% to 93% in the

population included in this study. So we can say that nor-

mal healthy individual has wide range of platelet aggre-

gation. No subject had 0% or 100% Platelet aggrega-

tion. So there will be some or more Aggregation re-

sponse by the platelets in each individual. And that is

determined at the time of conception in the form of

Prakṟti.

CONCLUSION

Platelet aggregation depends on Prakṟti. Prakṟti can be

used as an objective tool for prognosis of Atherosclerotic

changes. Assessment of Prakṟti is an important parameter

to decide the line of treatment of anti-platelet drugs or

advise to avoid the atherosclerotic changes. Medical ge-

netics seeks to understand how genetic variation relates

to human health and disease. So now from this study we

can say that assessment of Prakṟti is a measure to judge

human health, disease proneness & drug response in an

individual.

Further scope of the study:

Although this work is done in a scientific way however

there are certain limitations of time and facilities availa-

ble.

1. Majority of volunteers are doing their graduation or

post-graduation. There may be variation in result, if vol-

unteers of other occupations will be included.

2. All the volunteers are from age group 16-40 years.

There is age related impairment in platelet aggregation,

so study can be done specifically including old age vol-

unteers.

ΛΛΛΛ

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Source of Support: Nil.

Conflict of Interest: None declared

ΛΛΛΛ

How to cite this article: Pimpalkhare et.al,. : The Aggrega-

tion response of Platelets in different Prakṟti. AAMJ 2017;

6:1600 – 1604.

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