Internationally indexed journal

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1 Internationally indexed journal Indexed in Chemical Abstract Services (USA), Index coppernicus, Ulrichs Directory of Periodicals, Google scholar, CABI,DOAJ, PSOAR, EBSCO, Open J gate, Proquest, SCOPUS, EMBASE,etc. Rapid and Easy Publishing The International Journal of Pharma and Bio Sciences (IJPBS) is an international journal in English published quarterly. The aim of IJPBS is to publish. peer reviewed research and review articles rapidly without delay in the developing field of pharmaceutical and biological sciences Indexed in Elsevier Bibliographic Database (Scopus and EMBASE) SCImago Journal Rank Impact factor 2.958*..

2 ISSN Chemical Abstracts Service ( CODEN IJPBJ2 Elsevier Bibliographic databases (Scopus& Embase) SNIP value 0.77 SJR IPP SNIP Source normalised impact per paper SJR SCImago Journal rank IPP Impact per publication Source (Powered by scopus (ELSEVIER) And indexed/catalogued in many more university *Instruction to Authors visit For any Queries, visit contact of

3 Research Article Pharmacology International Journal of Pharma and Bio Sciences ISSN EVALUATION OF LOCAL ANAESTHETIC AND ANTIPYRETIC ACTIVITY OF TERMINALIA CHEBULA RETZ IN EXPERIMENTAL ANIMAL MODELS RAJYALAKSHMI.N 1, NAGESH RAJU.G 2 AND RAMANATH.B 2* 1 Department of Pharmacology, Rajiv Gandhi Institute of Medical Sciences, Srikakulam, Andhra Pradesh, India. 2 Department of Pharmacology, Basaveshwara Medical College, Hospital and Research Center, Chitradurga, Karnataka, India. ABSTRACT Dry fruit of Terminalia chebula is used in leprosy, anemia, narcosis, piles, chronic, heart disease, diarrhea, anorexia, cough and excessive secretion of mucus. The present study was undertaken to evaluate the local anesthetic and antipyretic activity of T.chebula in experimental animal models. Ethanolic extract of T.chebula (EETC) was tested in local anesthetic action by (i) intracutaneous wheal in guinea pigs and (ii) plexus anesthesia in frogs. In both the models, 2% xylocaine was used as the standard drug. Brewer s east induced pyrexia in rats before and after treatment of EETC compared with paracetamol. The test drug in concentrations of 10% and 20% produced 70.56% and % anesthesia respectively by the intracutaneous wheal compared to 97.64% anesthetic effect produced by 2% xylocaine (P<0.001). The mean onset of anesthesia with the test drug was min compared to min (P<0.001) for the standard drug in the plexus anesthesia model. In the anti-pyretic model, EETC in doses of 400 and 600 mg produced dose-dependent reduction in mean temperature at various hours of observation. The present study shows that EETC has significant local anesthetic and antipyretic activities. KEY WORDS: local anaesthetic, Antipyretics, Terminalia chebula. RAMANATH.B Department of Pharmacology, Basaveshwara Medical College, Hospital and Research Center, Chitradurga, Karnataka, India. *Corresponding author P - 219

4 INTRODUCTION Medicinal plants are part and parcel of human society to combat diseases, from the dawn of civilization. Herbal medicines are in great demand in the developed as well as developing countries for primary healthcare because of their wide biological and medicinal activities, higher safety margins and lesser costs. 1 Terminalia chebula is a plant species belonging to the genus Terminalia, family Combretaceae. 1 It is commonly known as Chebulic myrobalan (Harra in Hindi). 2 The Sanskrit name Haritaki is rich with meaning, referring to the yellowish dye (harita) that contains the god Siva (Hari, i.e. the Himalayas) and that it cures (harayet) all the diseases. 3 It is moderate to large sized tree found throughout India, chiefly in deciduous forests and areas with light rain fall, but occasionally found in moist forest up to an altitude of 1500 meters. 4 Terminalia chebula has been extensively used in ayurveda, It is a component of the classic Ayurvedic combination called Triphala (three fruits). 1 The dried ripe fruits have traditionally been used in the treatment of asthma, sore throat, vomiting, hiccup, bleeding piles, gout, heart and bladder diseases. Its paste with water is found to be anti-inflammatory, analgesic and having purifying and healing capacity for wounds. 5 It is given as adjuvant herb in chronic fever. 1 It has been used to treat various ailments like hemorrhoids, dental caries, bleeding gums and oral ulcers, diarrhoea, gastroenteritis, malabsorption syndrome, vesicular and renal calculi, neuropathy, paralysis, memory loss, epilepsy, depression, diabetes, tumors, skin diseases, as well as intermittent fever, rheumatism, arthritis, gout, etc. 1,5 The plant is reported to have antibacterial, antifungal, antiviral, antioxidant, hepatoprotective, cardioprotective, antidiabetic, hypolipidemic, antispasmodic, and various other activities. 1 A survey of literature revealed that no scientific study on the local anaesthetic and antipyretic activities has been reported on the fruits of the plant. The present study was designed to evaluate the local anaesthetic and antipyretic activities of the fruit extracts of the Terminalia chebula Retz. MATERIALS AND METHODS Plant Collection The fruits of Terminalia chebula were purchased from a noted and authenticated Ayurvedic shop during the month of febraury Preparation of extract The fruits of Terminalia chebula were shade dried and reduced to coarse powder in a mechanical grinder. The powdered materialobtained was then subjected to successive extraction by hot percolation method using ethanol solvents in a soxhlet extractor. The extract obtained was evaporated at 45 C to get a semisolid mass. The extracts thus obtained were subjected to phytochemical analysis. The percentage yield of alcoholic extract was found to be 36.50%w/w and the extract was used for further studies. The study was performed between the months of february2014 and March 2014, in the department of pharmacology, Basaveshwara Medical College and Hospital, Chitradurga, Karnataka. Phytochemical studies Phytochemical studies revealed the presence of tannins like chebulic acid, chebulagic acid, chebulinic acid, corilagin, gallic acid, gallotannins and ellagic acid; fructose, amino acids, succinic acid, ascorbic acid, flavonol glycosides, alkylamide, triterpenoids, coumarin, betasitosterol, resin and anthroquinone. 6 Animals Fully grown male guinea pigs (300 to 400 g), frogs, and albino rats ( g) were procured from the central animal house of the institute. The animals were housed at controlled room temperature (242 0 C; relative humidity 60-70%) in a 12 h light-dark cycle. They were given standard laboratory diet and water ad libitum. The experimental protocol was approved by the Institutional Animal Ethics Committee (Reg.no.1284/ac/09/CPCSEA). P - 220

5 Drugs The following chemicals were used: xylocaine (AstraZeneca), sodium chloride (Triveni chemicals), Ethyl alcoholic extract of fruit of Terminalia Chebula (EETC) Local Anesthetic Activity 1. Intracutaneous wheal in guinea pigs The animals were divided into four groups [Table 1]. On the day prior to the study, the hair on the back of guinea pigs near the midline (four different areas of 4 cm each) were clipped and removed. The drugs were injected intracutaneously in equal volumes of 0.2 ml into the shaved areas and wheals were marked with ink and the time of injection noted. The normal responses of the animals were observed first by applying pin pricks in the midline. Six pin pricks were then given uniformly every five minutes at an interval of four seconds on the wheal areas. The responses were recorded up to 30 min. A localized skin twitch, usually accompanied by a squeak, was considered as the normal response to pin prick. When the animal failed to respond either by twitching of the muscle or squeaking following a pin prick, a negative response was recorded. 7, 8 {Table-1} Tabel 1 Local anaesthetic activity of Ethanolic extract of Terminalia chebula on intracutaneous wheal in guinea pigs. Group Drug No. of Negative response Mean % failure of response Control 0.9% Normal Saline Test 10% EETC * 70.56* Test 20% EETC * 87.52* Standard 2% Xylocaine * 97.64* * P<0.001 when compared to control; n=6 in each group. GRAPH 1 P - 221

6 2. Plexus anesthesia in frogs The frogs were divided into three groups [Table 2]. They were damaged brain and spinal cords with the help of pithing needle. The abdominal viscera were excised and removed through a transverse incision made just below the sternum thereby forming a pouch. The lumbar plexus was exposed carefully without damaging it. The frogs were pinned to vertical boards with their legs hanging down. The drugs were administered into the abdominal pouch in sufficient volumes to submerge the lumbar plexus. The left and right limbs of the frogs were immersed every minute for a maximum period of 10 s in beakers containing 0.1(N) HCL and normal saline, respectively. Afterwards the feet were rinsed in water. The time taken by the animals failing to withdraw their feet was recorded as 9, 10, 11, 12, the "onset of local anesthetic action." 13 {Table-2} Tabel 2 Local anaesthetic activity of Ethanolic extract of Terminalia chebula on plexus anesthesia in frogs. Group Drug Onset of local anaesthetic action (meansem) (min.) Control 0.9% Saline Test 20% EETC * Standard 2% Xylocaine * * P<0.001 when compared to control; n=6 in each group. GRAPH 2 3. Anti-pyretic activity Brewer s yeast-induced pyrexia Rats were divided into four groups of six animals each. Fever was induced in rats by subcutaneous injection of 20 mg/kg of 20% suspension of Brewer's yeast in normal saline below the nape of the neck.10 Initial rectal temperature was recorded. After 18h, animals that showed an increase of C in rectal temperature were selected. The control group (Group I) rats received saline solution (0.9% w/v, NaCl) 2 ml/kg p.o., standard group (Group II) received Paracetamol 150 mg/kg p.o. 10, 11, Group III and IV received 400 mg/kg and 600mg/kg of EETC p.o. respectively. Antipyretic activity of EETC was assessed by measuring the rectal temperature with thermometer at 0, 30, 60, 120 and 180 minutes following drug administration. 14 P - 222

7 Table 3 Effect of Ethanolic extract of Terminalia chebula fruit on Brewer s yeast-induced pyrexia in Rats. Groups Drugs I Control II Paracetamol (p.o) III IV EETC (p.o) EETC (p.o) Rectal Temperature ( 0 C ) Dose mg/kg 0 min 30 min 60 min 120 min 180 min Values are mean S.E.M. (n=6) Significance vs. control group: **p<0.05. GRAPH 3 ** ** **37.86 ** ** ** ** ** **37.37 Blue = Control, Red = Test (Paracetamol), Green = EETC 400mg/kg, Violet = EETC 600mg/kg Statistical Analysis The results were analyzed for statistical significance by one-way ANOVA followed by Dunnet's't' test. A 'P' value of < 0.05 was considered significant. RESULTS In an intracutaneous wheal model in guinea pigs, the test drug in a concentration of 10% and 20% produced 70.56% and 87.56% anesthesia compared to 97.64% anesthetic effect produced by 2% xylocaine. The P - 223

8 negative responses of the standard and test groups showed a highly significant increase when compared to the control group. An increased concentration of the test drug produced an increase in local anesthetic activity (r=1) [Table 1]. The mean onset of anesthesia with the test drug was min compared to min for the standard drug. The onset of local anesthetic activity in the test and standard groups was significantly different from the control group [Table 2]. The anesthetic action of the standard and test drugs continued till 30 min of our observation period. In the antipyretic model, EETC in doses of 400, 600, mg/kg reduced the temperature of pyretic rats significantly from the first hour to third hour, respectively (P<0.05). Paracetamol lowered the temperature significantly throughout the observation period [Tabel-3]. DISCUSSION The local anesthetic activity of EETC was studied by the methods described by Burn et al., with slight modifications on (i) intracutaneous wheal in guinea pigs and (ii) plexus anesthesia in frogs. In the present study, 2% xylocaine was used as the standard drug in both the models, whereas the above workers used nupercaine as the standard in the intracutaneous wheal model and cocaine in the plexus anesthesia model. The wheal model is suitable for estimating the degree of anesthesia and its duration simultaneously, whereas the plexus anesthesia determines the onset of anesthesia. 13 The mean onset of local anesthetic action with EETC in concentration of 20% was min (P<0.001). The anesthetic action continued till 30 min of the observation period. The findings suggest that EETC possesses a significant local anesthetic property. The antipyretic activity was studied by a yeast-induced method of Brownlee. For the production of pyrexia, yeast is widely used. Various workers used different concentrations and different doses of yeast. The mean initial basal rectal temperature in this study was to 38.56º C. The rise in temperature after 3 h of induction was to 37.37º C. In this study, EETC reduced temperature of pyretic rats significantly from first hour to the third hour, respectively. Preliminary phytochemical studies showed the presence of flavonoids. There are reports that some flavonoids are predominant inhibitors of either cyclooxygenase or lipo-oxygenase. 6 Flavonoids are hydrolyzed by saliva to deliver aglycones that have protective effect in the oral cavity. Some workers have reported that T.chebula contains alkylamides. 15 The local anesthetic property of EETC observed in the study could be due to the presence of alkylamides. The antipyretic activity of T.chebula demonstrated in the present study could be due to the presence of flavonoids. CONCLUSION The present study concludes that the ethanolic extract of Terminalia chebula (EETC) has local anaesthetic and antipyretic activity in guinea pig 10%, 20%; frogs 20%, and rats at the doses of 400mg/kg and 600 mg/kg. However, this is a preliminary study and further study needs to be carried out for knowing the possible mechanism of actions. REFERENCES 1. Chattopadhyay RR, Bhattacharyya SK. Terminalia chebula: An update, Pharmacognosy Reviews, 1(1): (2007). 2. Verma N, Singh AP, Amresh G, Sahu PK, Singh A, Mishra N. Review on wonderful and miraculous Triphala. Journal of Pharmacy Research, 4(3): (2011). 3. Singh MP, Sharma CS. Wound healing activity of Terminalia Chebula in experimentally induced diabetic rats. Int.J. PharmTech Res. 3(2): (2008). P - 224

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