An Observational Study of Physical Properties of Blood Based on Dosha Dusti in Polycythemia

Introduction

Polycythemia vera is a stem cell disorder characterized as a panhyperplastic, malignant, and neoplastic marrow disorder with predominance of erythrocytes. Disease is characterized by impaired oxygen delivery due to sludging of blood, bleeding complications, thrombotic complications such as headache, dizziness, vertigo, tinnitus, visual disturbances, angina pectoris, intermittent claudication, epistaxis, gum bleeding, ecchymosis, gastrointestinal (GI) bleeding, splenomegaly (75% of patients), hepatomegaly, plethora and hypertension.¹ These features of Polycythemia vera mimics Shonita dusti laxana.

Among the saptha Dhatu, Rakta is responsible for Jeevana karma.² Most of the diseases are rooted with abnormal Rakta Dhatu. Detailed descriptions of the nidana and lakshana of Rakta dusti is explained in most of the literatures of Ayurveda.

The proper physiological functioning of the Raktha Dhatu depends upon the physical properties of Rakta. The changes in the physical properties of the Rakta in relation to dosha Kopa are also explained in the text books of Ayurveda. But, in the practice of Ayurveda, assessments of these normal and abnormal physical properties are not developed.

One of the tools for the diagnosis is to understand the changes in the physical properties of Dhatu based on the Doshic variations. Hence an attempt was made for assessing the physical properties of blood, based on Dosha dusti using various newly designed tests in Polycythemia.

Methodology

Source of data

The test for assessment of physical properties of blood on Dosha dushti was done at the SDM Research Center for Ayurveda and Allied Sciences, Udupi.

Samples were collected from O.P.D and I.P.D of S.D.M Ayurveda Hospital, Udupi.

Study design

Various special tests were done on collected blood samples by assessing the characteristics of Raktha based on classical and contemporary parameters. The test findings were noted on special proforma prepared with all points of physical properties of blood, based on Dosha dushti as mentioned in Ayurveda and allied sciences, along with various laboratory investigations. Inclusion criteria considered were diagnosed cases of Polycythemia with Hb% more than 16.5 gm/dL in males and 16 gm/dL in females, patients of either gender, patients in the age group of 20 to 60 years. Pregnant and lactating mothers and other chronic systemic illness were excluded from the study.

Study population

Participants were divided into a control group consisting of 30 healthy individuals and a test group consisting of 30 patients diagnosed with polycythemia.

Pilot study

A pilot study was conducted on a small sample of normal healthy individuals to set the diagnostic parameters planned for the study. The normal parameters of these newly designed tests were decided based on the results obtained in the pilot study.

Following the pilot study, the same tests were conducted on patients diagnosed with polycythemia to assess raktadushti by various doshas depending upon the observations made.

Assessment criteria

Assessment of blood was done based on Lakshanas mentioned in Brihatrayi which are listed in Table 1, 2 and 3, by conducting various tests with the help of contemporary parameters.

The assessment parameters of blood have been developed based on classical features of doshadushti using contemporary parameters. The assesement criteria with observations is mentioned in Table 4.

Varna - Colour

Blood is red in colour due to the convergence of hemoglobin present in it. The hemoglobin protein is comprised of subunits called hemes, which contain metal iron. When the iron atom gets oxygenated, it becomes red, and when the iron is deoxygenated, its begins to get darker in shade.⁶ Similarly, varna/ shade of raktha was surveyed by Pratyaksha pramana pariksha (direct perception). Based on the shade of blood, separate doshadushti was summarised as per the modern parameter of haemocheck meter. The other colours mentioned in classics like harita and neela were perceived directly (No samples other than red shades were received during the course of the study).

Assessment criteria:

The colour of the blood was assessed soon after withdrawing. A drop of blood was put over the strip of Haemocheck (Hemoglobin shading scale) which indicates the haemoglobin value based on the colour of blood (Figure 1a). Thus doshic contribution was observed by considering colour of blood as assessment criteria.

Gandha – Odour

This was assessed through perciveing the odour of the blood immediately after the collection of blood ghranendriyapariksha to rule out foul smell (puti gandha) of pitta dushita rakta.

Sheethaushnaguna - Temperature

Blood within the body equilibrates at all times to the body's core temperature. Classical references suggest that the temperature of rakta will vary based on doshadushti involved.

Assessment criteria: The temperature of the blood was assessed immediately after the collection from venepuncture using a Multi Stem Thermometer with an external sensing probe as shown in Figure 1b.

Skandana property - Bleeding and Clotting time

Clotting time

Clotting time is the time required for a sample of blood to coagulate in vitro under standard conditions.

Assessment criteria: In this study, clotting time of blood was assessed by capillary tube method. The blood was collected by venipuncture with the help of a syringe. Blood was filled into the glass capillary tube immediately. The capillary tube was then rolled in between palms to maintain proper temperature. The capillary tube was tilted 45° to check the progress of clotting and the tube was broken at intervals of 30 seconds till the fibrin thread was observed. The time taken for fibrin thread formation was noted.

Bleeding time

The test for bleeding time is dependent on appropriate functioning of platelets. Blood vessels and platelets affect the earliest hemostasis. The assessment was done by recording the time taken to stop the bleeding after the superficial skin was punctured.

Assessment criteria: In this test, the ear lobe was punctured with a lancet and the bleeding spot was collected on a Whatman filter paper at an interval of 30 seconds till bleeding stopped and the total duration was noted.

The normal range of bleeding time was 2 to 8 mins and normal clotting time was 7 to 12 mins.⁷

Bahala, picchila, mamsapeshiprabha - Hyper viscosity

In this test, soon after collection of blood, 20 µL of blood was immediately allowed to flow from the upper edge of the elongated cylindrical glass jar and then to flow down till it reached the bottom. The method followed is shown in Figure 1c.

Assessment criteria: Time taken for flow down to reach the bottom of the flask was noted.

Sheegragaami andmandhagaami - Spreading nature

The spreading nature of the blood is the total area covered by blood in a particular duration. In this test, 20 µL of blood was dropped immediately over the Whatman filter paper (Figure 1d).

Assessment criteria:

a) Area covered by the blood after spreading for 30 seconds.

b) Area covered after complete drying along with time taken to dry was noted.

Phenilam - Frothing nature of the blood

About 2 mL of blood was taken in a plain test tube, and subjected to linear shaking movements per second, for a duration of 30 seconds. Formation of froath was noted and length of the same froath was measured using measuring scale (Figure 1e).

Snighdam, rooksham, parusham - Triglyceride

The snigdhta and rookshata of blood were assessed by evaluating the concentration of triglycerides present in the blood. Samples showing increased triglyceride levels were compared with snigdha quality of blood.

Vishadam - Serum transparency

In this test, the collected blood was subjected to centrifugation and the serum was assessed for transparency and turbidity through inspection as shown in Figure 1f. The same sample was subjected to a color absorption test using a spectrophotometer at 590 nm wavelength.

Results

Results were assessed and analyzed using one way ANOVA, followed by Dunnet’s Multiple ‘t’ test and Post Hoc test. Statistical data with results are summarized in Table 5.

Discussion

Varna - Colour

The difference in the shade of the blood between the test group and the control group was statistically significant. The majority of the test group samples exhibited dark red to blackish red shading. It is suggestive of raktadushti by pitta and vatadosha. According to classical literatures of Ayurveda, shyava, neela, krishnavarna is found in pitta and vatadushita rakta. Hence, dark red to blackish red is a marker of high hemoglobin levels, according to hemocheck shading comparator.⁸

Gandha - Smell

As per the classics, rakta vitiated by pittadosha emits visragandhata (putrid smell). In both study groups, no specific smell character was seen.

Sheethaushnaguna - Temperature

The temperature of the blood between both the groups was statistically non-significant. As per the Ayurvedic literature, the pittadushtarakta will be atyushna. It was observed that some samples in the test group had slight increase in blood temperature than the normal. However, the findings were not statistically significant.

Skhandana property - Bleeding time

This test was performed to assess the askandi and chirasravi feature of dushtarakta and to clarify the predominance of kapha and pittadosha over raktadushti.

The raktadushti by pitta shows askandi property which indicates delayed clotting time and raktadushti by kaphadosha shows chirasravi features indicating delayed bleeding time.

In present study, none of the patients showed delayed bleeding and clotting time.

Bahala, picchila, mamsapeshiprabha - Blood viscosity

This test was performed to assess the hyperviscosity of blood and snigdha, ghana, bahala, picchila properties of shonitadushti. The prakupita kaphadosha vitiates the rakta responsible for abnormality in its qualities and exhibits features such as atisnighda, picchila, mamsapeshiprabha and Ghana.

Viscosity is the quality that describes a fluid's resistance to flow. So the hyperviscosity of blood shows greater resistance to flow pressure and thus the slope of the flow-pressure relationship is less steep.

The present study estimated the values in individuals categorized into test and control groups. Test performed on both the groups was statistically significant, the values of the test group were slightly higher than the control group, signifying hyperviscosity of blood.

With the above evidence of blood hyperviscosity / snigdha, picchila, bahala qualities of rakta, raktadushti by kapha and pittadosha can be inferred.

Sheegragaami and mandagaami - Spreading nature of blood – assessing total area of spread- time taken for drying

In the present study, it was found that the spreading nature of blood between the test and the control groups was statistically highly significant.

It was evident that the extent of spreading of blood in majority of test group samples was diminished compared to control group. Increased blood thickness (hyperviscosity) could be the reason for decreased spread.

The rakta which is vitiated by vatadosha will show sheegragaami - extended spreading nature.

But in the present study, majority of the polycythemic patients’ blood did not show increased spreading zone of blood over Whatman paper compared to control group.

Only three patients from study group showed increased spread area, suggestive of raktadushti by vata and these patients were diagnosed to have vatavyadhis. Among them, one patient had pakshaghata and two patients had sandhigata vata. Both were belonging to older age group where vatadosha is dominant.

Rest of the test group samples showed reduced spreading area compared to control group, which may be because of raktadushti by kapha and pittadosha signifying styaya, ghana, picchila, bahalaquality industarakta.

The relation between the time taken for drying of blood in both the groups

Most of the patients’ blood from the study group demonstrated delayed drying and it even took more than five minutes in some smaples. Such a manifestation is due to hyperviscosity of blood. Hence, raktadushti by kaphadosha can be inferred where blood shows atisnigdha, Ghana and picchilata.

Phenilam - Frothy nature of blood

This test was done to assess the phenila character of blood. The dooshitavata vitiates rakta and as a result, frothy appearance in blood will be observed along with other features. Test performed in both the groups was statistically non-significant. Hence, it can be inferred that raktadushti by vatadosha is not evident in Polycythemia.

Snighdam, rooksham, parusham - Triglyceride levels

This test was done to assess the snigdha property of blood. The present study estimated the values of serum triglycerides in individuals categorized to test group and the control group.

However, a significant number of test group samples showed higher values of triglycerides compared to control group. It was suggestive of raktadushti by kaphadosha indicating increased snighdata, picchilata in blood.

Vishadam - Serum transperancy

This test was performed to assess the vishada (clear-less-dense) quality of rakta. Test was performed with the help of spectrophotometric principles to measure the quantity of light-absorbing capacity of serum samples.

The aggravated vatadosha vitiates rakta and produces vishadata in rakta, which can be considered as thinner/ clear/less-dense serum.

The present study estimated the values of colour absorbance capacity of serum at 590 nm, in individuals categorized to the test and control groups. Test performed between both the groups was statistically insignificant. With this, it is clear that Raktadusti by Vatadosha is not evident.

Conclusion

Health research has a high value in society in managing health care. Improving health care for the future is the focus of health research. This research helps doctors and researchers to learn about human health and diseases. It also helps to learn better ways to diagnose, prevent and treat diseases.

Hence in this study, new technical objective parameters were developed to assess the physical properties of rakta. They are haemocheck strip for colour, multi stem thermometer for temperature, cylindrical glass jar for hyperviscosity, plain collecting tube for frothy nature, capillary tube for clotting time, Whatman filter paper for bleeding time and spreading nature, concentration of triglyceride for assessment of snigda, ruksha, parusha properties of blood, observation of transparency and turbidity for assessing vishada property and ghranendriya pareeksha for the assessment of gandha.

Objective parameters such as physical properties of blood are suggestive of Raktadushti by Pitta and Kaphadosha in patients under study. It was evident that among the various newly designed Raktapariksha assessing the doshic vitiation on Rakta, statistically significant results were observed for blood colour, spreading nature (reduced spreading because of styaya, bahala, ghana, picchila guna), and blood viscosity (snigdha, picchila).

Though the rest of the parameters did not show statistical significance, certain observational findings showed satisfactory similarities with the lakshana enlisted under Raktadushti. Absence of serum transparency (snigdha), increased triglyceride levels (atisnighdata), frothy nature (phenilam), bleeding time and clotting time (askandi) and temperature (ushna) were the abnormal properties of rakta suggestive of raktadushti by pitta and kaphadosha in patients under study.

Ethical Clearence

Ethical clearence approval has taken from Institutional Ethical clearence Committee of Shri Dharmasthala Manjunatheshwara Educational Society, Ujire with approval no : SDM/ACA-49/ECH23/17-18.

Conflicts of Interest

None