International Journal of Medical and Health Sciences

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1 International Journal of Medical and Health Sciences Journal Home Page: ISSN: Original article Haematological studies of Emblica officinalis formulation on Wistar Rats Narendra Kumar Savala* 1, Parvathaneni Nagasri Haritha 2 and Mangamoori Lakshmi Narasu 3 1 Research Scholor,CBT, IST, JNTUH, Hyderabad, Andhra Pradesh, INDIA 2 Assistant Professor of Cardiology, GMC, Guntur, Andhra Pradesh, INDIA 3 Professor,CBT, IST, JNTUH, Hyderabad, Andhra Pradesh, INDIA ABSTRACT Over three-quarters of the world population relies mainly on plants and plant extracts for health care. More than 30% of the entire plant species were used for medicinal purposes. Emblica officinalis is one of the widely used food supplement for the prevention of many ailments. A comprehensive account of the effect of Emblica officinalis formulation on some of the haematological parameters in wistar rats is presented in this paper. The whole blood samples drawn from orbital plexus were collected in K2 EDTA Vacutainers and were subjected for haematological evaluation in haematology cell counter, SYSMEX-KX 21 and the parameters studied are Total white blood cell count (WBC), Red blood cell count (RBC), Haemoglobin (Hb) gm/dl, Haematocrit (HCT), Mean corpuscular volume (MCV), Platelets. The results showed that there are no haematological abnormalities. In addition there is a better haematological status of the treated rats with respect to control rats. However, further investigations are suggested to identify the active components present in the Emblica officinalis formulation and to elucidate the mechanism at molecular level. KEYWORDS: Emblica officinalis formulation, Wistar rats, Haematology, Haemoglobin, Haematocrit, White Blood Cells, Red Blood Cells, Mean Corpuscular Volume, platelets INTRODUCTION Traditional systems of medicine continue to be widely practised on many accounts. Population rise, inadequate supply of drugs, prohibitive cost of treatments, side effects of several allopathic drugs and development of resistance to currently used drugs for infectious diseases have led to increased emphasis on the use of plant materials as a source of medicines for a wide variety of human ailments. The world Health Organization has approved traditional medicine as a part of treatment for primary health care needs [1]. Among known higher plant species approximately 5000 species have specific therapeutic value. These are classified according to their botanical classification and geographical distribution [2]. Among the all estimated plant species, only 6% have been studied for biological activity and about 15% have been investigated phytochemically [3]. Various drugs have entered the international market through exploration of ethno pharmacology and traditional medicine [4, 5]. One such plant with therapeutic value amla has been Int J Med Health Sci. April 2012,Vol-1;Issue-2 29

2 explored for its role in DNA damage and repair The plant contains tannins like glucogallia, corilagin, chebulagic acid and 3, 6-digalloyl glucose. Root yields ellagic acid, lupeol, quercetin and β-sitosterol [6]. The fruits of Amla are used in many medicinal preparations of Ayurvedic and Unani systems of medicine as well as food supplement [7, 8]. The effect of crude amla (traditionally known as emblica officinalis formulation) and total serum protein and its fractions was studied in rabbits and the results showed that the total protein levels and increased body weight indicating the positive nitrogen balance and having anabolic effect without affording resistance against diseases [9]. As many drugs alter the physiological status in the body of any animal including human beings, it is essential to study the crucial haematological parameters. Hence, the present study focused on the haematological parameters to understand the variations of physiological status of the animal. MATERIALS AND METHODS: Collection of Emblica officinalis formulation: Emblica officinalis formulation was procured from kottakkal ayurvedic vaidya Sala, Kerala. Emblica officinalis formulation is available as a rejuvenating agent in Ayurvedic practice and is prepared in combination of amla, ghee and honey. The rasayana is a mix of amalaki powder (12gm), ghee (12gm) and honey (24gm) - together packaged as 45gm/day. As per the ayurvedic literature, it should be one thola (about 11.66gm) to be taken for one mandala (45 days). Actual amalaki (Processed amla powder) would then be about 0.2gm/kg/day assuming they are 60kg in weight. Procurement of Animals for Study Animals were procured from National Institute of Nutrition (NIN), Hyderabad and cohorts of Wistar strain rats inbred over generation and maintained in animal house and they were used for the following studies. Study Design In this study, 48 animals were subjected for an additional supplement of Rasayana and 48 were on normal diet. These two groups were designated as Treated and Control, respectively. Rats were maintained in a pathogen free environment with a 12hrs light-dark cycle. Food and water were provided ad libitum. Randomization and Numbering of animals: After randomization, all the animals were acclimatized to laboratory conditions for a minimum of five days, and assigned to four groups of 24 animals consisting 12 males and 12 females. The females were nulliparous and nonpregnant. Study consists of 96 rats (48 males + 48 females) which were divided into four groups and each with 24 rats. Among the four groups first was sacrificed on 3rd month, second, third and fourth groups were sacrificed in 6th, 9th and 18th months respectively. Randomization ensured that the allocation of treatments to animals/groups was independent of their characteristics and were similar in all the groups. It was also taken care while randomization, base variables were homogenized and were allotted to different groups. The need for randomization applies not only to the allocation of the animals to the different (control as well as treatment) groups but also anything that can materially affect the recorded response. Dosage and Route of Administration Administration of Emblica officinalis formulation is within the standard volume of 10 ml/kg body weight. Emblica officinalis formulation was administered at the concentrations of 0.45gms/1ml per wistar rat which is 6 times greater than the human dosage and reason behind this is the higher metabolic rate of wistar rats is 6 times greater than human, and this dosage has been determined keeping the concentration prescribed for human dosage as 0.75 gms/kg body weight. Wistar rats 6-8 weeks of age, weighing 120 to 165 g of both sexes have been used and the animals used in this study were obtained from NIN. Route of administration and reason for choice Oral route has been chosen because it is the proposed route of administration to humans. The dosage can be accurately administered. The Oral route is one of the proposed routes for toxicity testing. The test item was administered by oral route. Haematology The whole blood samples drawn from orbital plexus were collected in K2 EDTA Vacutainers Int J Med Health Sci. April 2012,Vol-1;Issue-2 30

3 and were subjected to haematological evaluation in haematology cell counter, SYSMEX-KX 21. The parameters selected for study are total white blood cell count (WBC), Red blood cell count (RBC), Haemoglobin (Hb), Haematocrit (HCT), Mean corpuscular volume (MCV), Platelets. Data Analysis The data of the above haematological parameters was subjected for statistical analysis using MS Excel The difference between the groups was tested. Independent t test is applied. RESULTS Blood parameters Values do not show much significant difference during 3,6,9 and 18 months except Platelets count with test statistic in males and in females. Net result is showing steady values and the platelets values are in the normal range and does not show any significance after 3rd month onwards. Table 1: Complete haemtological profile for the control and amla induced male rats. Males Parameter Control and treated Months 3 values 6 values 9 values 18 values HB(gm/dL) C-M 14.73± ± ± ± T-M 14.25± ± ± ±0.9 HCT% C-M 46.93± ± ± ± T-M 45.10± ± ± ±3.0 MCVμ 3 C-M 55.87± ± ± ± T-M 56.30± ± ± ±2.8 PLTX 10 3 C-M ± ± ± ± T-M ± ± ± ±154.5 RBCX10 6 C-M 8.41± ± ± ± T-M 8.01± ± ± ±0.7 WBCX10 3 C-M 14.85± ± ± ± T-M 16.78± ± ± ±3.6 Values are shown in mean ± standard deviation Int J Med Health Sci. April 2012,Vol-1;Issue-2 31

4 Table 2: Complete haemtological profile for the control and amla induced female rats Females Parameter Control and treated Months 3 values 6 values 9 values 18 values HB(gm/dL) C-F 14.18± ± ± ± T-F 13.72± ± ± ±0.9 HCT% C-F 42.80± ± ± ± T-F 41.68± ± ± ±3.5 MCVμ 3 C-F 57.63± ± ± ± T-F 56.88± ± ± ±1.5 PLTX 10 3 C-F ± ± ± ± T-F ± ± ± ±83.4 RBCX10 6 C-F 7.43± ± ± ± T-F 7.33± ± ± ±0.5 WBCX10 3 C-F 7.77± ± ± ± T-F 10.87± ± ± ±1.4 Values are shown in mean ± standard deviation HB - Haemoglobin HCT - Haematocrit, Corpuscular Volume, PLT - Platelets WBC - White Blood Cells, RBC - Red Blood Cells, MCV- Mean DISCUSSION HB, HCT, MCV, PLT, RBC and WBC levels following the Emblica officinalis formulation treatment are within the normal levels. Table 1 shows the haematological data for males and females for all the 4 groups i.e. control vs. treated Males and Females during various life stages. i.e. after 3, 6, 9 and 18 months of dosage. The results reveal that there is a normal haematological profile in all the parameters of both males and females. Oral administration of Emblica officinalis formulation did not produce any adverse effect on haematological parameters of animals. In the Haemoglobin levels of control versus treated among the males, there is an initial decline in the 3rd month of dosage and a slight increase in the 6th month and significant enhancement of haemoglobin levels were observed in 9 and 18 months of dosage in relative to the controol. It may be because of the adoptation of the physiological system for the additional supplement and regained these levels at the later stages of life. These Haemoglobin levels are also similar to the females except where females had regained it little faster and showed slight increase in the haemoglobin levels at the 6th month of dosage itself in comparison to the controlled rats. Int J Med Health Sci. April 2012,Vol-1;Issue-2 32

5 Haematocrit levels are increased at 9th month and decreased levels during 3,6 and 18 months of dosage and decreased at the 3rd month in females where there is increased levels are observed during the remaining dosage testing periods. Mean corpuscular values in the treated females are lower than the control females during all stages of life where as in males there is no particular pattern observed. In the platelet levels of control versus treated among the males, there is an initial decline in the 3rd month of dosage and a significant enhancement of haemoglobin levels were observed in 6, 9 and 18 months of dosage in relative to the control while there is no such pattern in female rats.rbc levels have not shown any pattern between control versus treated rats, whereas WBC levels are better in comparison to control rats. CONCLUSION: Haematological parameters of treated versus controlled are within the normal limits indicating health status of subjects at the haematology level. Based on the results obtained it can be concluded that Emblica officinalis formulation, a herbal preparation boosts haematological response. However, further investigations are needed to identify the active components present in the Emblica officinalis formulation and to elucidate the mechanism at molecular level, which can pave way for rational design for effective molecules for its use as a therapeutic agent in DNA damage and associated health problems. ACKNOWLEDGMENTS Narendra Kumar Savala is grateful to the Council of Scientific and Industrial Research (CSIR) for senior research fellowship and Jawaharlal Nehru Technological University Hyderabad for PhD (Full Time) fellowship. We thank Prof. Y. Prameela Devi for discussions and help. REFERENCES: 1. Goyal, R.K,.Investigation of Cellular and Molecular Mechanisms for Anti-diabetic Drugs With special Reference to unani and Ayurvedic Herbal Medicines. In:Traditional system of Medicine, Abdin, M.Z. and Y.P. Abrol (Eds). Narosa publishing House, New Delhi 2005 pp: Joy, P.P., Thomas, J., Mathew, S., and Skaria, B.P.. Medicinal Plants. Tropical Horticulture Vol. 2. (eds. Bose, T.K., Kabir, J.,Das, P. and Joy, P.P.). Naya Prokash, Calcutta, 2001 pp Verpoorte, R., R. van der Heijden, H.J.G. Ten Hoopen and J.Memelink,. Metabolic Engineering of plant secondary metabolic pathways for the production of new chemicals. Biotechnol.Lett : Goyal, R.K. and S.S.patel,.From concept to Reality in Herbal Medicine: Zingiber officinale as potential antidiabetic Herbal Drug with specific 5-Hydroxytryptamine Receptors as The target. In: Recent Advances in Herbal Drug Research and Therapy, Ray, A.and K.Gulati(Eds.).I.K.International publishing House pvt. Ltd., New Delhi, 2010 pp: Cooper, E.L.,. Complementary and alternative medicine, when rigorous, can be science. Evid. Based Complement. Alternat. Med., 2004, 1: Thakur, R.S.; Puri, H.S.; Husain, Akhtar: Major Medicinal Plants of India.. Central Institute of Medicinal and Aromatic Plants, Lucknow, India Kirtikar, K.R. and B.D. Basu,. Indian Medicinal Plants. International Book Distributors, Dehradun, India, Int J Med Health Sci. April 2012,Vol-1;Issue-2 33

6 8. Mishra, P., M. Verma, V. Mishra, S. Mishra and G.K. Rai,. Studies on development of ready to eat amla( Emblica officinalis) chutney and its preservation by using class one preservatives. Am. J. Food Technol., 2011, 6: Tiwari A; Sen,S. P; Guru, LV; The effect of Amalaki (phylanthus embalica) rasayana on biologic system. J. Res. Ind., Med., 1968; 2 : 189. *Corresponding author: Narendra Kumar Savala snk2018@yahoo.co.in Int J Med Health Sci. April 2012,Vol-1;Issue-2 34