ANTIASTHMATIC POTENTIAL OF THE ROOTS OF ECHINOPS ECHINATUS ROXB. DR.P.L.RAJAGOPAL* DR.K.PREMALETHA** DR.K.R.SREEJITH *** INDHU ANNIE CHACKO**** K.V. NIMISHA**** * Professor and Head, Dept. of Pharmacognosy and Phytochemistry, Academy of Pharmaceutical Sciences, Pariyaram Medical College, Kannur, Kerala **Professor, Principal and Head, Dept. of Pharmaceutics, Academy of Pharmaceutical Sciences, Pariyaram Medical College, Kannur, Kerala. ***Assistant Professor, Dept. of Pharmaceutical chemistry, Academy of Pharmaceutical Sciences, Pariyaram Medical College, Kannur, Kerala ****Final Year Bachelor Degree in Pharmacy (B.Pharm), Department of Pharmacognosy and Phytochemistry, Academy of Pharmaceutical Sciences, Pariyaram Medical College, Kannur, Kerala ABSTRACT The present study aims at the evaluation of the ethanolic extract of the roots of Echinops echinatus Roxb.on invitro animal model. Histamine induced contraction in isolated goat tracheal chain showed that ethanolic extract of the roots of Echinops echinatus Roxb. inhibited the contractile effect of histamine P<0.05. A dose dependent contraction of goat tracheal chain was observed during the evaluation. In addition the root contain Lupeol which can cause reduction of eosinophils, which may also contribute the aniti asthmatic action of the plant. KEYWORDS: Antiasthmatic, Antihistamine, Lupeol, Echinops Echinatus INTRODUCTION Asthma is a disease characterized by bronchial airway inflammation resulting in increased mucous production and airway hyper responsiveness. Around 100 to 150 million people are suffering from asthma in the world. The most worrying aspect of these statistics is that the numbers are escalating phenomenally and the deaths from this condition have reached over 180 million annually. 1 India has estimated to be 15-20 million of asthmatics. Genetic predisposition is one of the factors in children for the increased prevalence. Other factors like urbanization, air pollution & tobacco smoke contributes more significantly. However there is no complete remedy to cure asthma. Further several classes of synthetic drugs have been adopted in the treatment of asthma. In many conditions the patient has to be administered with drugs for a prolonged period or even lifelong. Administration of these drugs for long period may result in adverse effects and chronic toxicities or drug- drug interactions. Therefore there are several attempts to explore the possibility of natural drugs 141
for asthma treatment. 2 Keeping this in view, a field survey was conducted to identify an herbal remedy for the management of asthma. Echinops echinatus (Asteraceae) is an herbaceous plant, widely distributed in desert regions of Kerala and Tamilanadu. The root, leaves, fruit and bark are most commonly used parts 3,4. In traditional medicine most of the diseases have been treated by administration of plant or plant product. Echinops echinatus Roxb is the useful traditional medicinal plant in India. Each part has some medicinal property. During the last five decades, apart from the chemistry of the Echinops echinatus Roxb compounds, considerable progress has been achieved regarding the biological activity and medicinal applications of Echinops echinatus Roxb. It is now considered as a valuable source of unique natural products for development of medicines against various diseases and also for the development of industrial products. The survey revealed the fact that Echinops echinatus was traditionally used in asthmatic conditions by the tribals of Marayur forest of Idukki district, Kerala State. 5,6 Since the antiastmatic potential of this plant was not claimed, the present study was aimed to screen the ethanolic extract of the roots of Echinops echinatus for its in vitro anti-asthmatic activity. MATERIALS AND METHODS Plant collection The Plant material was collected from the hilly area of Marayur, Idukki district of Kerala state in the month of September and the botanical identity of the same was confirmed and a voucher specimen bearing No. EeRoxb 837 was deposited at the Pharmacognosy department of Academy of Pharmaceutical Sciences, Pariyaram Medical College, Kannur, Kerala. Preparation of extracts The roots of Echinops echinatus were dried in shade, powdered and sieved. The powdered material was extracted with ethanol.the extract was evaporated under reduced pressure using rotary evaporator until all the solvent has been removed to give an extract sample. Phytochemical Screening procedure. 7 Preliminary phytochemical screening has been carried out as per the standard 142
ANTIASTHMATIC SCREENING Isolated goat trachea chain preparation Isolated adult goat tracheal tissue was obtained immediately after slaughterhouse of the animals. Trachea was cut into individual rings and tied together in series to form a chain. Trachea was suspended in bath of Krebs solution and was continuously aerator at 37+0.5 C. Dose response curve of histamine in plane Krebs solution and in 800 µg/ml ethanolic root extract of Echinops echinatus act in Krebs solution was taken. Graph of percentage of maximum contractile response on ordinate and concentration of histamine on abscissa was plotted to record dose response curve of histamine, in absence and in presence of drug extract. 8,9 Statistics The statistical analysis was performed by using One way analysis of variance (ANOVA) followed by Dunnett s test for individual comparison of groups with the control. The p values less than 0.05 were considered as significance. Result and Discussions Phytochemical Screening Phytochemical screening revealed the presence of saponions, flavonoid glycosides, poly phenolic compounds in the alcoholic extract. Antiasthmatic screening It was observed that ethanolic root extract of Echinops echinatus inhibits contraction produced by histamine in these tissue preparations. Histamine (10µg/ml) was taken in different dose level and dose response curve was plotted. Study revealed that the root extract of Echinops echinatus exhibits significant (p<0.01) percentage decreased contraction at concentration 800 µg /ml in goat tracheal chain preparation Dose dependent response relationship was observed during the evaluation. Histamine contracts the trachea-bronchial muscle of guinea pig, goat, horse, dog and man. 13 143
Table - 1 Effect of root extract of Echinops echinatus root on histamine induced contraction on isolated goat tracheal chain preparation Sl No Concentration of Histamine Percentage Maximum Response (Mean ± SEM) (10µg/ml) Control Test 1 1 36.57 ± 1.31 17.22 ± 1.01* 2 2 39.39± 1.17 29.65 ± 1.30** 3 4 48.46 ± 2.15 38.49 ± 1.27** 4 8 57.35 ± 2.13 46.65 ± 1.41** 5 16 78.57 ± 2.29 58.09± 1.09** 6 32 99.46 ± 2.88 62.24 ± 1.20** n = 6 Values are in Mean ± SEM. Control - D.R.C. of Histamine in the absence of Echinops echinatus root extract. Test - D.R.C. of Histamine in the presence of Echinops echinatus root extract Statistical analysis done by using Student s t -test. **p<0.01, significantly different from control. For the evaluation of antiasthmatic studies isolated goat tracheal chain is selected because it is very easy to handle and prepare and also when compared with the guinea pig the sensitivity of the goat tracheal chain is much higher. In the present study of isolated goat tracheal chain preparation, there is a right side shift of dose response curve of histamine in the presence of ethanolic root extract of Echinops echinatus indicating antiasthmatic action. The plant seems to be having a promising role in the treatment of bronchial asthma could be due to the presence of flavonoid glycosides 12 and steroidal saponins which can inhibit the release of several mediators such as serotonin, prostaglandins and histamines by inhibiting the biosynthatic pathways of inflammatory mediators. The effects of the alcoholic root extract may be because of presence of phytochemicals such as saponins 10 flavonoids, and tannins, known to possess similar effects. In addition the plant also contains lupeol. 6 Lupeol can cause the reduction of cellularity and eosinophils in the broncho alveolar lavage fluid. Treatment with lupeol can also reduce the production of mucus and overall inflammation in the lung. Thus lupeol attenuates the alterations characteristics of allergic airway inflammation. 11 The investigation of the mechanisms of action of this molecule may contribute for the development of new drugs for the treatment of asthma. 144
Acknowledgements The authors are thankful to the Principal and Staff of Karaveli College of Pharmacy, Mangalore, Karnataka and Academy of Pharmaceutical Sciences, Pariyaram, Kannur, Kerala for providing facility for the research work. References 1. Alagesaboopathi C. Ethnomedicinal plants their utilization by villagers in kumaragiri hills of salem district of Tamil nadu. Afr J Trad CAM. 2009;6(3):222-7. 2. Bhangare NK, Pansare TA, Ghongane BB,Nesari TM. Screening for Anti-Inflammatory and Anti- Allergic activity of Bharangi {Clerodendrum Serrratum (Linn.) Moon} in Animals. Int J Pharm Bio Sci. 2012; 3(4):245 254. 3. Castillo JC, De-Beer EJ. The Tracheal Chain-l - A Preparation for the Study of Antispasmodics with Particular Reference to Bronchodilator Drugs. J. Pharmacol. Exp. Ther., 90, 1947: 104-109. 4. Chaudhari KN, Lahiri SC. Role of Goat Trachea for an Isolated Tracheal Chain Preparation. Indian J. Pharmacology, 6(3), 1974:149-151. 5. EhrightT, Chua S, Lim D J, Pulmonary eosinophilia syndromes, Ann. J. Allergy, 1989; (62):277-283. 6. Kokate CK.Practical Pharmacognosy, 4thedn, Vallabha Prakashan, New Delhi,1999;149-156. 7. Nesamony S, Medicinal Plants, State Institue of languages, Trivandrun, 1998:(2)52. 8. Qureshi R Floristic and ethnobotanical study of Desert-Nara Region, Sindh. Shah Abdul-latif University, Khairpur, Sindh, Pakistan, 2004; 227-231. 9. Rajani GP, Patil UA, Deb J Anti asthmatics -an overview. Indian drugs. 2006;43(10):781-9. 10. Rajani GP, Ranjan KM, Mahesh NK, Thimmaiah NV, Faiyaz A, Rajamma AJ. Asthma, an overview, herbs to treat asthma. J Pharm Res. 2009;2(7):1188-91. 11. Singh S et.al, Flavonoids from Echinops echinatus, Journal of Asian Natural Products Research, 2006:8(3);197-200. 12. Sofia Eram et.al.,experimental evaluation of Echinops echinatus as an effective hepatoprotective, Academic journals, 2013;(8)39:1919-1923. 13. Vasconcelos JE, Teixeira MM et.al, The triterpenoid lupeol attenuates allergic airway inflammation in a murine model, International Immunopharmacology, 2008:8(9);1216-1221. 145