A A M J Anveshana Ayurveda Medical Journal

A A M J Anveshana Ayurveda Medical Journal www.aamj.in ISSN: 2395-4159 Research Article Immunomodulatory activity of Polyherbal drugs Bhringarāja - āsava and Gudūci Satva : A clinical comparison Anjali B Prasad 1 Sachin Agrawal 2 Sheetal Rothe 3 K Shankar Rao 4 A b s t r a c t Introduction: Impaired immunity can be co-related with Rājayakśma mentioned as the prime of all disease in Ayurveda. Ayurveda states two types of immunity (Vyādhikśamatva) i.e. resistance to disease (Vyādhi-virodhitavam) and strength to arrest the progress of disease (Vyādhi-pratibandhakatvam). The present study enables us to evaluate the drugs critically and scientifically so that their immunomodulatory potential is validated. Aims: To compare and study the immune-modulatory activity of Bhringarāja-āsava and Gudūci Satva. Material and Methods: An open Study on 60 immune compromised patients was carried out. Gudūci Satva, Bhringarāja-āsava and both were administered in three different groups of randomised 20 individual in one group for the assessment on subjective (appetite, digestion, exercise capacity and sleep were assessed) and objective parameters (T.L.C., D.L.C., E.S.R., Hb%, SGOT, SGPT, Total bilirubin, Direct bilirubin and Indirect bilirubin, IgG, IgE and routine and microscopic urine examinations). The data was elicited before and after the trial of 45 days. Statistical analysis used: The subjective parameters were subjected to Wilcoxon's MPSR (matched pair signed rank) test and the objective parameters were assessed by student s t- test (paired) for their statistical significance. Results: Group C showed better immunomodulatory activity than group A (Gudūci Satva) and group B (Bhringarāja-āsava). Key words: Vyādhikśamatva, Vyādhi-pratibandhakatvam, Bhringarāja-āsava, Gudūci Satva, Rājayakśma. 1 Assistant Professor, Agada Tantra Evam Vyavahar Ayurveda, 2 Assistant Professor, Rasashastra and Bhaishajya Kalpana SGCAS & Hospital, Sriganganagar, Rajasthan, 3 Ayurvedic Physician, Sumankanta Child & Adult Hospital, Navi Mumbai, Maharashtra 4 Professor & Head, Dept. of Rasashastra & Bhaishajya Kalpana, NIA, Jaipur CORRESPONDING AUTHOR Dr. ANJALI B PRASAD Assistant Professor, Department of Agada Tantra Evam Vyavahar Ayurveda, SGCAS & Hospital, Sriganganagar, Rajasthan. Email: anjali.prasad75@gmail.com AAMJ / Vol. 2 / Issue 3 / May June 2016

INTRODUCTION A yurveda The science of life aims to protect health of healthy individual and treat the various ailments. [i] Both of these aims are directly associated with Immunity of the individual. Immunity refers to resistance of the body to pathogens and their toxic products. It can be classified as Innate immunity and Acquired immunity. Innate immunity (Inborn genetic or constitutional make up) Non- specific Specific (Species, Racial, Individual against a particular organism) Acquired immunity (acquired during life) Active Acquired immunity (Natural, Artificial) Passive Acquired immunity (Natural, Artificial) Mechanism of Innate immunity 1. Mechanical barrier (Provided by Skin and Mucosa) 2. Surface secretion ( Saliva, Gastric Juice, Tears, Sebaceous secretion) 3. Humoral defence mechanism (Lysozyme, polypeptide, Compliments, Interferons) 4. Cellular mechanism of defence (Phagocytes, Natural Killer cells, Eosinophils) Acquired immunity: The resistance that an individual acquires during his life time is known as acquired immunity. It is antigen specific and may be antibody-mediated or cell-mediated. It is of two types Active and Passive. Active immunity: Acquired by synthesis of antibodies (Humoral immunity) and production of immunocompetent cells (cell mediated immunity) by the individuals own immune system in response to antigenic stimulation. Active immunity can be induced naturally (from subclinical or clinical infection) and artificially (by inducing antigens). Passive immunity: It is administered to a recipient in a readymade form. Natural passive immunity is acquired in foetus IgG are transferred from mother placenta; after birth Immunoglobulins pass through breast milk. Human colostrums are rich in IgA which are resistant to digestion. By active immunization of mother during pregnancy the immune status of the neonate can be improved. Artificial passive immunity is administered to the recipient by injecting readymade antibodies by administration of hyper immune sera. [ii] Multiple defence mechanisms protect an individual from microorganisms, harmful, factors, according to modern medical science. Thus all the factors or the state in which, a person remains free from disease, could be said to bear on immunity. The concept of immunity was described by ancient Ayurvedic Aacharya s as Vyādhikśamatva which prevents the development of disease as well as resist the further progression of development of disease. The level of Vyādhikśamatva is not equally maintained in all types of constitutions, which means it varies from individual to individual, regardless to their similar nutritional, environmental, physical and mental status. [iii] Aacharya Chakarapani has mentioned Vyādhikśamatva as Vyādhi-virodhitavam and Vyādhi-pratibandhakatvam. Vyādhi-virodhitvam: Power to resist the occurrence of disease. Vyādhi-pratibandhakatavam: Strength to arrest the progress of disease. [iv] Vyādhi-virodhitvam can be co-related to innate immunity and Vyādhi-pratibandhakatvam can be related to acquired immunity. Immune system of body consists of Antigen, Antibodies, and specifically sensitized cells. Antigen is a substance that produces Antibodies, which interact specifically with it. It may be introduced in body or may be formed within the body in response to bacteria, toxin, foreign body etc... An antibody (Ab), also known as an immunoglobulin (Ig), is a large Y-shaped serum protein that is produced by plasma cells that is used by the immune system in response to counteracting a specific antigen. The immunoglobulins (Igs) of which there are five classes; IgG, IgE, IgM, IgA, IgD, protect an individual against invasion by infection of various bacteria, virus, organisms, etc. Today it is believed that we have extended defence mechanism E.g. Immunological surveillance limits the development of the tumour cells, malignant cells. Impaired immunity can be simulated with Rājayakśma the king of all disease. It is also called as Kshaya. The Bala vridhikar yoga is most useful in treatment of Rājayakśma. In Gada Nigraha and Bharat Bhaishajaya Ratanakar. Bhringarāja-āsava [v] and in A.F.I. part I [vi] (Yogaratnakar Rājayakśma chikitsa) [vii] Gudūci Satva is recommended for the treatment of Rājayakśma. AAMJ / Vol. 2 / Issue 3 / May June 2016 692

IgG and IgE are directly associated with body s defence mechanism. Thus these two Immunoglobulins were considered for this study to see the Immunomodulatory effect of Bhringarāja-āsava and Gudūci Satva. Aims and objectives:- To understand the concept and mechanism of immunity. To validate the immunomodulatory activity of Bhringarāja-āsava and Gudūci Satva. MATERIAL AND METHODS Ingredients of Bhringarāja-āsava and Gudūci Satva were identified by the experts of Dravyaguna dept. and were prepared in dept. of Rasa Shastra and Bhaishajya Kalpana, NIA, Jaipur. Preparation of Bhringarāja-āsava was three step procedures pūrvakarma (Preoperative procedure), pradhān karma (Operative procedure) and paśchāt karma (Post operative procedure). Pūrvakarma: It includes collection of the raw drugs preparation of all factors required for preparation of Āsava; Powdering of drugs: All drugs (Harītaki and the Prakśepa dravya) were shadow dried and made into fine powder mess size 120 after removing foreign material; Preparation of Swarasa: Bhringarāja Panchānga was dusted well to remove the external impurities, soaked overnight in eight times R.O. water. Next day Soaked Bhringarāja Panchānga was kept for cooking on Mandāgni till it got reduced to one fourth of its initial volume; [viii] then prepared Swarasa was strained through cotton cloth and Harītaki powder and half of the Jaggery was added into it and remaining half amount of the jaggery was added after fermentation process was completed, to avoid delay in fermentation process. Dhūpana (fumigation) and Lepana (Coating): Fumigation of the container was done with some Rakśaghna dravyas (Haridra, Kapur, Ghee, Vacha, Jatamansi, Chandana, Guggulu, Vidanga) and the inner surface of the vessels [ix], [x], [xi] was coated with ghrita. Pradhān karma: All ingredients were mixed well and kept in 20 lit food grade fumigated Plastic containers. The transparent containers were used as it was easy to observe fermentation process in them. Remaining half of Jaggery was added into the samples after completion of the fermentation, this Jaggery doesn t take part in the process of fermentation and due to which thickness of Āsava was increased i.e. like syrup form, it becomes palatable to patients due to sweet taste. Paśchāt karma: Filtration was done, before adding ½ Jaggery to get clear, thin Āsava. Prakśepa and remaining jaggery was added to the thin Āsava. Packing and sealing was done properly in 500ml food grade bottles. [xii] Table No. 1 showing ingredients of Bhringraj-āsava S.No Sanskrit Name Scientific Name Family Part used Quantity 1. Bhringarāja Eclipta alba Asteraceae Wh.pl. 12.250lts. 2. Harītaki Terminalia chebula Combretaceae Fr. 384gms. 3. Pippali Piper longum Piperaceae Fr 96gms. 4. Jātiphala Myristica fragrance Myristicaceae Fr 96gms. 5. Lavanga Sygizium aromaticum Myrtaceae Fl. 96gms. 6. Twak Cinnamomum zeylanicum Lauraceae Br. 96gms. 7. Elā Elatteria cardamom Zingiberaceae Sd. 96gms. 8. Tamālapatra Cinnamomum tamala Lauraceae Lf. 96gms. 9. Nāgakesara Messua ferrea Guttiferae Fl 96gms. 10. Guda Cane jaggery (Old) -- 9.600 kgs. Gudūci Satva preparation Gudūci was procured and Gudūci Satva was prepared at Chitrakut karvi dham, Satana (MP). Matured Gudūci stem was used so as to get more yields; Gudūci stem was washed with tap water and wipe out with cotton cloth to remove dust and foreign particles adherent to it. Then upper cover of Gudūci stem was removed and cut into pieces get clear starch. Then Gudūci stem was crushed well up to fibers. Then it was mixed with 21 parts of water and soaked overnight. Next day stem was rubbed well within water and washed until stickiness of material disappears, then material was strained and kept in flat AAMJ / Vol. 2 / Issue 3 / May June 2016 693

dish for sedimentation. After sedimentation process upper liquid layer was removed by decantation process and the sediment was collected and washed seven times to get fresh Satva. Satva was packed in airtight bottle as it is very hygroscopic in nature to prevent absorption of moisture. [xiii] Clinical valuation of Bhringarāja-āsava and Gudūci Satva: Clinical trial enables us to explore and validate the drug scientifically. It is a powerful tool which when used properly can help Ayurvedic formulations to get a strong hold in today's era of Evidence Based Medicine'. Present study is randomised single blind trial. The drugs were allocated to different groups by using simple random sampling (lottery method). Selection of cases Source the patient for present study were screen out from the OPD and IPD of Rasashastra Department, N.I.A. Hospital, Jaipur (Raj.). Age Group patient of any age group irrespective of their sex, occupation and living standards. Inclusion Criteria History of more than 3-5 episodes of following feature in last year or recurrence of these symptoms, such patients are included in inclusion criteria for clinical study. URTI LRTI Mild fever/chronic fever Diarrhoea Weakness Weight loss Allergic to any component. Exclusion Criteria Acute and sever episode of infections. Infection associated with congenital heart diseases (IHD). Specific pathology such as malignancies. Grouping and Posology Grouping: There were 3 groups for clinical study, each group consist minimum 20 patients each, 78 patients were registered for clinical trial, out of which 18 patients were dropped out for irregularity in treatment and follow up. So excluding these data of 60 patients is considered for clinical study. Table No. 2 showing grouping and Posology of the drug. Particulars Group A Group B Group C No. Of Patients 20 20 20 Drug Gudūci Satva Bhringarāja-āsava Gudūci Satva + Bhringarāja-āsava Dose 1gm b.d. 15ml b.d.. 1gm b.d. Gudūci Satva + 15ml Bhringarāja-āsava Duration Max. of 45 days Max. 45 days Max. 45 days Follow up Fortnightly Fortnightly Fortnightly Anupāna With honey With equal quantity of water Both water and honey The diagnosis was made on the basis of clinical findings and laboratory investigations. Laboratory investigations The Laboratory Investigations were done at the Central Laboratory, N.I.A Hospital, Jaipur Hematological Investigations : T.L.C., D.L.C., E.S.R., Hb% Immunological Investigations : IgG, IgE Urine Examinations : Routine and microscopic. Study Design Type of Study Randomized single blind trial Randomization Simple random sampling (lottery method). Method of sevana Vātātapika (outdoor regimen) Apathya / contraindications Salty and sour food, spicy and irritant food Initially the trial medicine of 15 days was given to the all registered volunteers. In case of Group A volunteers, Gudūci Satva capsules (60 capsules) and Group B volunteers 500ml of Āsava is given, in Group C Gudūci Satva capsules (60 capsules) and 500ml Āsava was AAMJ / Vol. 2 / Issue 3 / May June 2016 694

given. The trial / treatment were continued up to 45 days with the follow up after interval of 15 days. Assessment criteria The patients enrolled for the study were assessed on subjective and objective parameters on the basis of preforma design for the study. The data was elicited both before commencement and completion of the trial by recording general observations, clinical parameters and values of pathological and bio-chemical investigations. Statistical Methods The subjective parameters were subjected to Wilcoxon's MPSR (matched pair signed rank) test and the objective parameters were assessed by student s t-test (paired) for their statistical significance. A level of 'P' <0.01, 'P' <0.001 and P <0.0001 were considered as statistically significant, highly significant and extremely significant respectively. Level of significance was noted and interpreted accordingly. Clinical observations (subjective parameters) As the present study was for only 45 days, the subjective parameters were designed with Sannikriśta Lakśanas (which manifests in a short period like changes in general health condition e.g. appetite, digestion, sleep, etc.); while Viprakriśta Lakśanas (which manifests remotely like Varna, Medha, Smriti and Ayu). The overall effect of these drugs on health was determined by clinical parameters, pathological and bio-chemical investigation. General observations Various demographic parameters viz, age, sex, religion/caste, occupation, socio-economic status, marital status of the volunteers was recorded. The patients were closely interviewed about their ages, sex, socio-economic status, marital status, religion, habitat, dietary habits, nature of job etc. and other relevant information as body weight, pulse rate, blood pressure, dietary habit etc. were noted. In this study out of 60 patients, it was found maximum patients (60 %) were in 21-30 years of age group; The sex ratio of females to males was 60% and 40% respectively; Maximum patients were Hindu (86.665) followed by Muslims (8.33%); 68.33% were married. Majority of the patients were from lower middle cases (53.33%); 91.4% of the patients were literate. Many patients had family history of allergy, recurrent infections, diabetes, hypertension etc. Maximum patients (38.33%) had chronicity of symptoms for 0-1 month. Treatment history: 41.66% patients were taking allopathic treatments; patients were vegetarian; 40% of patients were having Madhyam Kośtha; 40% Patients were assessed with Viśamāgni. Majority of the patients were suffering from Atinidra (38.33%); 51.66% patients showed vibandha; 38.33% patients were having vātakaphaja prakriti; 58.33% were rājasik-tāmasik prakriti ; 35% of patient were twaka sāra; 50% of patients showed avara Satva; madhyam vyāyām śaktii were found in 36.66%; 36.66% patients had madhyam Abhyavaharan śakti. OBSERVATIONS AND RESULTS Table No. 3 Haematological and Immunological changes in Group A, Group B and Group C Parameters Group A Group B Group C T P Results T P Results T P Results Body weight 8.34 >0.01 N.S 8.34 <0.001 H.S 8.34 <0.0001 E.S. Hemoglobin 1.08 >0.01 N.S 3.74 <0.001 H.S 4.17 <0.0001 E.S. TLC 1.18 >0.01 N.S 1.02 >0.01 N.S 0.12 >0.01 N.S Neutrophils 1.39 >0.01 N.S 0.19 >0.01 N.S 0.18 >0.01 N.S Lymphocyte 0.82 >0.01 N.S 0.12 >0.01 N.S 1.18 >0.01 N.S Eosinophils 1.10 >0.01 N.S 0.90 >0.01 N.S 0.4 >0.01 N.S Monocytes 2.37 <0.001 H.S 2.37 <0.01 S 2.37 <0.0001 E.S. ESR 2.56 <0.001 H.S 5.37 <0.001 H.S 1.43 <0.0001 E.S. SGPT 2.08 <0.01 S 4.36 <0.001 H.S 3.2 <0.0001 E.S. SGOT 6.27 <0.001 H.S 5.88 <0.001 H.S 2.68 <0.01 S. Total Bilurubin 0.43 <0.01 S 0.94 <0.01 S 0.72 <0.01 S. Ditect Bilurubin 4.71 <0.001 H.S 4.36 <0.001 H.S 4.71 <0.001 H.S Indirect Bilurubin 1.82 <0.01 S 1.39 <0.01 S 1.82 <0.0001 E.S. IgG 8.32 <0.0001 E.S 1.54 <0.001 H.S 1.88 <0.0001 E.S. IgE 6.20 <0.0001 E.S 4.14 <0.0001 E.S 6.75 <0.0001 E.S. S significant, N.S.- non significant, H.S. Highly significant, E.S. Extremely significant AAMJ / Vol. 2 / Issue 3 / May June 2016 695

Table No. 4 Subjective assessment of symptoms of Group A, Group B and Group C Symptoms Group A Group B Group C T P Results T P Results T P Results Weakness 4.58 <0.01 S 4.24 <0.01 S 16 <0.001 H.S. Loss of appetite 3.67 <0.01 S 11.0 <0.001 H.S. 4.18 <0.01 S Weight loss 3.16 >0.01 N.S. 9.57 <0.001 N.S. 14 <0.001 H.S Mild/chronic fever 10.25 <0.001 H.S. 3.16 >0.01 N.S. 4.18 <0.001 H.S. Diarrhoea 2.12 >0.01 N.S. 0.36 >0.01 N.S. 1.5 >0.01 S URTI 2.53 >0.01 N.S. 7.09 <0.01 S. 5.29 <0.01 S LRTI 3.16 >0.01 N.S. 11.08 <0.01 S. 13 <0.01 S Allergy 2.12 <0.01 N.S. 9.69 <0.01 S. 0.24 <0.01 S S significant, N.S.- non significant, H.S. Highly significant, E.S. Extremely significant DISCUSSION The patients who were suffering from URTI, LRTI, mild fever, chronic fever, diarrhoea, weakness, weight loss, allergic to any complaints, selected for the study. The patients were selected irrespective of their age, sex, religion, socio-economic conditions etc, through random, sampling. Thus the patients were randomly divided into three groups. Each group was having 20 patients. They were treated as Group A ( Gudūci Satva), Group B (Bhringarājaāsava) & Group (Gudūci Satva & Bhringarāja-āsava combine) with the clinically assessable common dose of 15 ml twice a day, i.e. in morning & night after food with equal amount of water as Anupāna and Gudūci Satva 1gm b.d. with honey for 45 days. Group A showed extremely significant results in IgG and IgE; highly significant results in monocytes, ESR, SGOT and direct bilirubin; significant results in SGPT, total bilirubin and indirect bilirubin; and insignificant results in body weight, haemoglobin, TLC, and neutrophils, lymphocytes and eosinophils. Group B showed extremely significant results in IgE; highly significant results in ESR, SGPT, SGOT, direct bilirubin and IgG; significant results in monocytes, total bilirubin and indirect bilirubin and insignificant results in TLC and neutrophills, lymphocytes and eosinophils in DLC. Group C showed extremely significant results in body weight, haemoglobin, monocytes, ESR, SGPT, indirect bilirubin, IgG and IgE; highly significant results in direct bilirubin; significant results in SGOT, total bilirubin; and insignificant results in TLC, neutrophils, lymphocytes and eosinophils. Group A showed highly significant results in mild/ chronic fever; significant results in weakness and loss of appetite; and insignificant results in weight loss, diarrhoea, URTI, LRTI and allergy. Group B showed highly significant results in loss of appetite; significant results in weakness, URTI, LRTI and allergy; while insignificant in weight loss, mild/chronic fever diarrhoea. Group C showed highly significant results in weakness, weight loss and mild/chronic fever; and significant results in loss of appetite, diarrhoea URTI, LRTI and allergy. CONCLUSION Gudūci Satva and Bhringarāja-āsava was very important in maintaining the immune status of the patients, also very effectively in improving symptomatic parameters and haematological and biochemical parameters. All patients tolerated medicines very well and no side effects or toxic effects of any of these drugs were reported by any of the patients, suggesting there by that the drug selected for the current clinical trial are absolutely safe for internal use by the patients. The patients who were dependent on Allopathic drugs had better improvement than those on Ayurvedic medicines. Finally the work of the results inferred that both Gudūci Satva and Bhringarāja-āsava boost immunity and can be used to enhance the resistance power of the patient. Over all, the present study suggest that the Gudūci Satva and Bhringarāja-āsava can be replaced when weakness, loss of appetites etc. are present but in case of fever, jaundice only Gudūci Satva was useful and in chronic cough, anaemia Bhringarāja-āsava was useful. So they cannot replaced by each other. But their combined use show better results. No complications were noted in any group. ^^^^ AAMJ / Vol. 2 / Issue 3 / May June 2016 696

REFERENCES i. Agnivesha, Charaka Samhita, Sutra Sthana 30/26, Vidyotini Hindi Commentary, Edited by Pt. Kashinath Pandey & Dr. Gorakhnath Chaturvedi, Reprint 2013, Chaukhambha Bharati Acadamy, Varanasi, pg-587. ii. Text book of Medical physiology By Indu Khurana; First print 2006; Reprint 2009; Published by Elsevier, a division of Reed Elsevier India Private Limited; Logix Park, A4 & A5 1 st Floor, Sector-16 Noida-201301, Uttar Pradesh. iii. Agnivesha, Charaka Samhita, Sutra Sthana 28/7, Vidyotini Hindi Commentary, Edited by Pt. Kashinath Pandey & Dr. Gorakhnath Chaturvedi, Reprint 2013, Chaukhambha Bharati Acadamy, Varanasi, pg-570. iv. Agnivesha, Charaka Samhita, Sutra Sthana 28/7, Ayurved Deepika commentary by Chakrapadidatta Edited by Yadavji Trikamji Acharya, Reprint 2009, Chaukhambha surbharti Prakashan, Varanasi, pg-178. v. Rasa Vaidya Nageendas Chaganlal Shah, Bharat Bhaishajya Ratnakar, Tritiya Part, Yog No. 4902, Reprinted 2012, B.Jain Publishers, New Delhi, Pg-653. vi. Ayurvedic Formulary of India, Part I, Edition-II 2003, Government of India, Ministery of Health & Family Welfare (English), New Delhi, pg-560. vii. Yogratnakar Rajyakshama chikitsa 14/1, Edited by Vaidya Laxmipati Shastri, Reprint 2010, Chaoukhamba Prakashan, Varanasi, pg-382. viii. Sharangdhara Mishra, Sharangdhara Samhita, Madhyam Khanda 1/4, Edited by Dr. Shailja Shrivastava, 4 th edition 2005, Chaukhamba Orientalia, Varanasi, pg-127 ix. Sharangdhara Mishra, Sharangdhara Samhita, Madhyam Khanda 10, Edited by Dr. Shailja Shrivastava, 4 th edition 2005, Chaukhamba Orientalia, Varanasi. x. Agnivesha, Charak Samhita. Chikitsa sthana14/168, Charaka Chandrika hindi Commentary, Edited by Brahmananda Tripathi, Reprint 1998, Choukhamba Surbharti Prakashana, Varanasi. xi. Acharya ShodhalaTripathi. Gada Nigraha. Chapter 6, Edited by Indradev Tripathi, 1969, Choukhamba Sanskrit Series, Varanasipp. pg.346-401. xii. http://www.ijaar.in/posts/images/upload/01_06_01.pdf xiii. http://www.iamj.in/posts/images/upload/547_554.pdf Source of Support: Nil. Conflict of Interest: None declared How to cite this article: Anjali et.al. : Immunomodulatory activity of Polyherbal drugs Bhringarāja - āsava and Gudūci Satva : A clinical comparison AAMJ 2016; 3:691 697 ΛΛΛΛ AAMJ / Vol. 2 / Issue 3 / May June 2016 697