European Journal of Cardiovascular Medicine

Siddha system of medicine is one of the ancient medical systems in India which reflects the life style and culture of the people (1). Siddhars, the founders of siddha medicine had designed the health practices including seasonal discipline and food regulation (2). Siddha system relays on the concept of “Food of Medicine” (1). Thus system prepares therapeutic drugs from green herbals. The herbal formulations serve as both drug and nutrient supplement (7). Panchabootham and uyir vayus are responsible for the organisation of this body and universe from the microcosm and macrocos (3,4). When the equilibrium of this panchabootham and uyir vayus are disturbed various disease are formed in our body (3,4).

Iron deficiency anaemia is a global public health problem. More than 700 millions persons in the world suffer from iron deficiency anaemia and about thrice the number from iron deficiency state (12,14). About the half of the population in the developing countries has iron deficiency anaemia. Infants and young children are particularly vulnerable to iron deficiency anaemia because their requirement for iron is high. Iron deficiency anaemia has been identified as a major health problem in India mostly affecting the pregnant women and young children (12,14).

Iron deficiency Anaemia is the state where in Iron content of the body is below normal, low transferring saturation and ferritin as well as high iron binding capacity (10,11). According to 2016 survey as per WHO, iron deficiency anaemia affects 1.3 billion people worldwide amongst which 43% are pre school children, 51% are pregnant woman and 37% are school aged children. Recent estimates of iron deficiency anaemia show that 52% of Indian women aged 15-49 years are anaemic (12). National Family Health Survey (NFHS) show prevalence of anaemia in 56.2 % of women of 15-49 year, 79.2% amongst children age 6-15 years,57.9% in pregnant women and 24.3% men age 15-49 year. Women are vulnerable part of society due to poor intake and absorption, increased requirements, menstrual loss and adolescent pregnancies (13).

Siddha system describes anaemia as paandu or veluppu noi, which means paleness. Anaemia is further classified into six sub types based on clinical symptoms. They are vali paandu, azhal paandu, iyya paandu, mukkutra paandu, nanju paandu and mannun paandu (6).

The symptoms of Iron deficiency anaemia are described as Azhal Paandu(6). siddha provides patient-friendly medication to overcome the anaemic condition.

As per siddha literature Thasadeepakkini Chooranam (TDC) plays a vital role in the management of anaemia (5). Although Thasadeepakkini Chooranam (TDC) has been playing a vital role in the management of anaemia without adverse effects.To enlighten the scientific basis for the use of Thasadeepakkini Chooranam (TDC) in Iron deficiency anaemia this study was done.

2.1 Drug selection:

The trial drug Thasadeepakini Chooranam was taken from the Siddha literature (5).

2.2 Ingredients:

1. Ferula asafoetida (Perungayam)
2. Acorus calamus (Vasambu)
3. Embelia ribes (Vaividangam)
4. Trachyspermum ammi (Omam)
5. Terminalia chebula (Kadukkai)
6. Plumbago zeylanica (Chithramoolam)
7. Rock salt (Induppu)
8. Saussurea costus (Kostam)
9. Piper longum (Thippli)
10. Cuminum cyminum (Seerakam)

2.3 Source of raw drugs:

The raw drugs for the preparation of the trial drug were procured from the raw drug shop at Tirunelveli, TamilNadu. The raw drugs are authenticated by the Pharmacognonist of Government Siddha Medical College & Hospital, Palayamkottai, TamilNadu.

2.4 Purification:

The ingredients of  Thasadeepakini Chooranam were purified as per authenticated Siddha literature (9).

2.5 Preparation of Thasadeepakini chooranam :

The ingredients were powdered in an iron mortar separately made into a fine powder and sieved it.Then the prepared Choornam (Powder ) was stored in a clean, air-tight glass container (5)

2.6 Evaluation of haematinic activity:

Animals :

The experiment was conducted in Wistar Albino rats of either sex, weighed 150-200g from the animal house of Kongunadu Arts and Science College, Coimbatore. The animals were maintained under standard laboratory condition with food and water ad libitum. The animals were allowed to acclimatize for 2 weeks before being subjected to experimental protocol. The animals were treated in line with the guide and care of laboratory animals as approved by the Institutional Animal Ethical Committee. IAEC No:PCP/IAEC/020/2016.Phenyl hydrazine (PHZ) used for induction of Anaemia and the standard drug Hematinic - Vit B12 syrup was purchased from authorized suppliers. 

Procedure

Induction of Anaemia :

 Animals were divided into five groups containing six animals in each. Phenyl hydrazine and haemotoxicity chemical is used to induce anaemia in rats .However, the result from animal health monitoring in the entire period of 14days showed no sign of morbidity and diseases .

The rats were randomly divided into five groups (6 rats per group) and treated daily for 4 weeks.Group I served as control and received regular rat food and drinking water ad libitum.Group II, III and IV, V rats received phenylhydrazine 10 mg/kg for 8 days orally to induce anaemia in rats. Rats that developed anaemia with haemoglobin concentration lower than 13 g/dl were recruited for the study. Tween 20 a vehicle received by group II. Group III received standard haematinic drug vit B12 (10 ml/kg). Group IV &V received Thasadeepakkini chooranam (500 mg/kg-1000mg/kg) diluted in a vehicle tween 20.

Treatment of the Animals :

 Group 2, 3, 4 and 5 animals are anaemic induced by phenylhydrazine. Group I (control) - received only water and food. These animals are not induced by anaemia. Group II (negative control) - received only vehicle Tween 20 (10 ml/kg) Group III (positive control) - received only Vit B12 syrup (1 ml/rat) a standard drug. Group IV & V -received 500 mg/kg-1000mg/kg of Thasadeepakkini chooranam respectively.

All administrations were by oral intubation. The absolute dose of test drug given to the rat was calculated by the body surface area ratio between human intended dosages against rat. All the groups were treated orally as single dose daily for three week.

The albino Wistar rats were healthy as shown by the normal appearance of general behavior, respiratory pattern, cardiovascular signs, motor activities, reflexes and normal change in skin fur. With regards to hematological values, most of values in treated groups were normal in comparison with the control group. Significantly, some values were different from those of the control group such as RBC, MCV, MCHC, and platelet. However, such values are within the normal ranges. These variations may have resulted from variation among animal groups (Feldman et al., 2000) (Inala et al., 2002). Therefore, these results suggest that the test drug did not cause hematological or immunological defects in rats.

Furthermore, blood chemical examination was performed in order to evaluate any toxic effects on liver. In this study, the levels of these blood chemical values were minor changes and remained within the normal range (Casley and King, 1980) (Levine, 1995) (Angkhasirisap et al., 2002).  Thasadeepakkinii chooranam sample given orally to Wistar rats did not produce toxicities.

Table – 1 Oral administration of Thasadeepakkini Chooranam extract on hematological parameters in Anaemic Induced rats

Values are expresses as Mean ± S.E.M.

All groups were treated with oral dose of 2g/kg body weight

No significant different from normal control

Haematological Parameters Analysis :

All the rats were fasted overnight and 0.5 ml of blood was collected on next day by puncturing Retro orbital sinus using capillary tube. The blood was collected after induction of anaemia with PHZ and during the end of first, second and third weeks of treatments. The red blood cell count (RBC), haemoglobin concentration (Hb), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and Packed cell volume (PCV) were determined using Haematogical analyzer HA - 22.

Statistical Analysis :

All the results were expressed as mean ± SEM of six animals. Analysis of variance was performed by ONE WAY ANOVA followed by Dunnet’s test. Probability values less than 0.01 were considered as significant.Oral administration of Phenylhydrazine for 8 days induced anaemia in rats. Thasadeepakkini Chooranam used in the management of anaemia. As a result there is an increase in the haematological parameters within 3 weeks compared to other group of rats.

Haematinic activity of Thasadeepakkini Chooranam in Phenyl hydrazine induced Anaemia in Rat Phenyl hydrazine caused significant decrease (P < 0.05) in Hb concentration, RBC count and haematocrit value in all rats indicating Anaemia. After the administration of test drug Thasadeepakkini chooranam , The hematological parameters are significantly increased (P <0.05). The PHZ - induced Anaemia was significantly reversed within one week of treatment with the test drug, reaching maximum by the third week. The effect of Haematinic syrup was comparable to the test drug Thasadeepakkini Chooranam .

Iron deficiency anaemia is a common disease affecting women especially in reproductive age (14). It is characterised by microcytic hypochromic RBC, in which MCV and MCH are reduced (10,11,15). It occurs due to defective haemoglobin synthesis. Iron in the body is used primarily for the synthesis of haemoglobin and normal erythropoiesis requires 20-25 mg of iron per day (10,11,15). Due to chronic blood loss, increased iron requirement, malabsorption of iron leads to iron deficiency (14,15). So administration of iron required to manage the disease without side effect. It can be prevented by increased dietary intake of iron and vit C for absorption. Administration of Phenylhydrazine a haemotoxicity chemical induce anaemia. Rats under the haemoglobin level 12 mg/kg are recruited for the study. The haematological parameters such as PCV, MCV, MCH and Hb are monitered for three weeks. After three weeks of treatment with Thasadeepakkini Chooranam the haematological parameters reach the normal levels.

In this study the oral administration of Thasadeepakkini Chooranam significantly increases the haematological parameters from first week of treatment. This study concluded the haematinic activity of Thasadeepakkani Chooranam was significant when compared to the standard haematinic drug.

1. Siddha System of Medicine - The Science of Holistic Health, Ministry of AYUSH, GoI. 2019
2. Dr. K. Durairasan, H. P. I. M, Noi illa Neri, 1993, 3rd edition
3. Dr. K. S. Uthamarayan, H.P.I.M., Siddha Maruthuvanga Churukkam.
4. Shanmugavelu.M, H.P.I.M, Noi Naadal Noi Muthal Naadal Thirattu- Part I&II
5. Kannusamy pillai.C. Sigitcha rathnadeepam -part-II 1993 8th Edition
6. Kupusamy mudaliyar, H.P.I.M, Siddha Maruthuvam, 2007 7th Edition.
7. Dr. Uthamarayan.K.S, H.P.I.M. Thottrakirama Araichiyum Siddha Maruthuva Varalarum.
8. Dr.Murugesamudaliar.K.S, Gunapadam Mooligai Vaguppu, 2002 2nd edition
9. Dr. Dheva asirvatham Samuel, M. D(S), Marunthu sei Iyalum Kalaiyum, Department of Indian Medicine and Homoeopathy
10. Sir Stanley Davidson, Davidson's principles and practice of medicine 22nd Edition
11. DAS P.C, DAS.P.K, Textbook of medicine, 2009 5th Edition.
12. De Benoist, B. Mclean E, Egli. l and Cogswell, M (2008) Worldwide prevalence of Anemia, WHO Global database of anemia.
13. World Health Organization, Anemia 2008.
14. "WHO Global Anaemia estimates, 2021 Edition". World Health Organization. Retrieved February 27, 2022.
15. Anemia: Practice Essentials, Pathophysiology, Etiology". November 9, 2021. Retrieved February 8, 2022.
16. Reactions of Haemoglobin with phenylhydrazine: A review of selected aspects. MD Shelter and MA Hill
17. Yeshoda KM: Phenylhydrazine anaemia in rats. Curr. sci 1942; 11: 360-363
18. Yadav AV, Undale VR : Evaluation of Haematinic activity of Naga Bhasma in phenylhydrazine induced Anaemia in experimental rats; Asian journal of Pharmaceutical and clinical research; vol 12, issue 12 Feb 2019
19. Sasipriya T, Evaluation of Haematinic activity of Kumaraveeriya Gaandha Chenduram in phenylhydrazine induced anaemia in experimental rats; World journal of Biology Pharmacy and Health Sciences, cross ref DOI: 10.30574/wjbphs