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IJBCP International Journal of Basic & Clinical Pharmacology Evaluation Print ISSN: 2319-2003 | Online ISSN: 2279-0780 IJBCP International Journal of Basic & Clinical Pharmacology DOI: http://dx.doi.org/10.18203/2319-2003.ijbcp20194127 Original Research Article Evaluation of dose dependent analgesic response by extracts of Myristica fragrans on albino wistar rats: an experimental study Imran Zaheer1*, Syed Ziaur Rahman2, Rahat Ali Khan2, Mehtab Parveen3, Priyamvada Sharma1 Department of Pharmacology, ABSTRACT 1F.H. Medical College and Background: The objective of the study was to evaluate analgesic activity of Hospital, Etmadpur, Agra, 2Jawaharlal Nehru Medical ethanolic extract, methanol and benzene fraction of Myristica fragrans on College, 3Department of wistar albino rats. Methods: The present study was carried out in the department of pharmacology Chemistry F/O Science, Aligarh Muslim University, Aligarh, JNMC AMU and F.H. Medical College, Agra. The analgesic activity was Uttar Pradesh, India evaluated by employing the Eddy’s hot plate method and tail flick response method. In both the tests, Rats of either sex weighing 150-200 g were used. The Received: 13 July 2019 total number of animals n=36 were allocated to six groups. Each group consist of six animals each. The response noted in animals that were tested by hot plate Accepted: 16 August 2019 method was reaction time for licking/biting of both the paws before and after administration of control & test drugs. However in Tail flick test, the pain *Correspondence to: threshold response was recorded before and after administration of control & Dr. Imran Zaheer, Email: imran82zaheer test drugs. The statistical analysis was done by using one-way ANOVA. The data is expressed as Mean±SEM. P<0.05 was considered to be statistically @gmail.com significant. Copyright: © the author(s), Results: Ethanolic extracts and methanol fraction of M. fragrans showed publisher and licensee Medip statistically significant (p<0.001) increase in reaction time for licking/biting in hot plate method. On the contrary a significant increase in pain threshold was Academy. This is an open- access article distributed under also recorded in tail flick response test. It is interesting to note that no the terms of the Creative significant degree of analgesia related to any dose of benzene fraction was observed. Commons Attribution Non- Commercial License, which Conclusions: The present study reveals the dose dependent significant permits unrestricted non- analgesic activity of the extracts of M. fragrans i.e. ethanolic extracts and methanol fraction in both the test. However, the degree of analgesia was commercial use, distribution, and reproduction in any recorded significantly higher in groups received higher doses of extracts of M. medium, provided the original fragrans. work is properly cited. Keywords: Analgesic activity, Myristica fragrans, Albino wistar rats INTRODUCTION gravity of adverse effects of NSAIDs it is necessary to search for new drugs with less adverse effects. In line Pain is a common subjective phenomenon which brings a with this many traditional medicinal plants have been patient to physician. It is associated with a number of used time to time for the development of new drugs with 4 diseases and is estimated that 80–100% of the population comparatively less ADRs as compare to NSAIDs. experience back pain at least once in the life time.1 Non- Recently, many natural medicines derived from steroidal anti-inflammatory drugs (NSAIDs) are the main medicinal plants, were considered as the effective and stay of treatment of pain.2 It is known fact that the risk of safer for the treatment of various diseases including 5 gastrointestinal bleeding and other side effects are inflammation and pain. One such medicinal plant associated with acute and chronic use of non-steroidal namely Myristica fragrans is claimed for it is valuable anti-inflammatory drugs (NSAIDs).3 Keeping in view the role in reducing pain and inflammation. Hence, it was www.ijbcp.com International Journal of Basic & Clinical Pharmacology | September 2019 | Vol 8 | Issue 9 Page 2134 Zaheer I et al. Int J Basic Clin Pharmacol. 2019 Sep;8(9):2134-2139 found worthwhile to evaluate the extracts of M. fragrans Central Animal House and were housed in polypropylene for their role in reducing pain in experimental animals. cages bedded with husk. They were provided with M. fragrans (Nutmeg) belongs to the family standard pellet diet (Ashirwad Industries, Chandigarh) myristicaceae, is one of the important spices used in and water ad libitum. The animal room was well- indigenous system of medicine in India. Its usefulness is ventilated and maintained under standard environmental reported in inflammations, cephalgia, helminthiasis, conditions throughout the experiment (temperature 18- halitosis, dyspepsia, flatulence, nausea, vomiting, 29⁰C, humidity 30-70%, 12 hour light/dark cycle). Rats diarrhoea, dysentery, colic, asthma, catarrh, neuralgia, were acclimatized to the laboratory condition for 1 week lumbago, stangury, amenorrhoea, menorrhagia, prior to experimental use. The study followed ARRIVE dysmenorrhoea, ulcers, liver and spleenic disorders, eye guidelines and was approved by the Institutional Animal diseases, impotency, skin diseases, freckles, cracks in Ethics Committee. feet, insomnia, delirium tremens, hyperdypsia, cardiac 6 disorders, fever and general debility. Earlier studies Acute toxicity study showed that ethanolic extract of M. fragrans have 7 analgesic activity. The present study is done to validate No toxicity of ethanolic extract of M. fragrans seeds was the earlier study and to screen additionally the effect of done as ethanolic extract of plant material is claimed to its methanol & benzene fraction. be safe. However, the limit toxicity of methanol and benzene fraction of M. fragrans in addition to acute METHODS toxicity in accordance to Organization for Economic Cooperation and Development (OECD) Guidelines 425” Plant materials was done on healthy adult female rats (100-150 g). While following the limit toxicity a dose of 2000 mg/kg was Seeds of M. fragrans were obtained from the local administered to a group of five animals for calculation of market. These were identified and authenticated by Chief LD50 of methanol fraction and they were observed for 14 Scientist, Raw Material Herbarium and Museum, days. However, no mortality related above limit toxicity National Institute of Science Communication and dose was noted.8 Information Resources (NISCAIR), New Delhi. A sample specimen of plant material was deposited in the Drugs NISCAIR bearing voucher number “NISCAIR/RHMD/2013/2345-125-2”. Pentazocine lactate (Inj. Fortwin, Ranbaxy Lab. Ltd., India). Preparation of extract Experimental protocol M. fragrans seeds were shade-dried. Plant material (100g) was powdered by using electrical grinder (REMI Analgesic activity by Eddy’s hot plate method auto-mix blender, Vasai, India). The powdered plant material was extracted with 300 ml of absolute alcohol by The present study was performed by using the Eddy’s hot employing Soxhlet apparatus. The plant extract was plate method (Orchid Scientifics, India). The hot plate is filtered, evaporated and dry at 40˚C on a water bath. Thus an electrically heated aluminium plate with a temperature the extract material (semisolid mass) was weighed to ranging between 55˚ to 56˚C. calculate its yield in percentage. The final yield of ethanolic extract of M. fragrans was 17.94%. Rats of either sex (150-200 g) were used. The response noted was reaction time of licking/biting of both paws. Preparation of fraction They were similarly screened and those responding in <6 sec were chosen. The paws of rats are very sensitive to One kg powder was made after crushing the dried roots heat at temperatures compared to skin. of M. fragrans. The air-dried powdered were exhausted with 95% ethanol and the solvent was isolated by steam The selected animals were placed on hot plate to record distillation. Under reduced pressure the extract was the response. The reaction time was measured at the concentrated to dark gummy mass. The residue so interval of 30, 60, 90,120,150,180, 210 and 240 minutes obtained was fractionated by refluxing in the consecutive after the administration of control and test drugs. order with benzene, ethyl acetate and methanol. The yield Propylene glycol 0.3 ml/100g p.o. served as control of methanol and benzene fraction of M. fragrans was whereas Pentazocine 30 mg/kg i.p. was administered as 4.17% and 5.31% respectively. standard drug. The cut-off time for response reaction was 30 seconds. The plate was wiped clean every time with Animals used saline if urination or defecation is found. Wistar albino rats weighing between 150-200 g of either The test was done in all 8 groups as shown in Table 1. sex were used. The animals were procured from the International Journal of Basic & Clinical Pharmacology | September 2019 | Vol 8 | Issue 9 Page 2135 Zaheer I et al. Int J Basic Clin Pharmacol. 2019 Sep;8(9):2134-2139 Table 1: Experimental design. maximum cut-off time of 15 sec adopted to prevent undue tissue damage. Tail flick latency was tested at 30 Groups Medication min interval for 4 hours. The test was done in 8 groups as Group I (Normal Propylene glycol 0.3 ml/100 g shown in Table 1. control) p.o. Group II Statistical analysis Pentazocine 30 mg/kg i.p (Standard control) Group III Ethanolic extract of M. fragrans The statistical analysis was done by using one-way (EEMF200) seed 200 mg/kg p.o. ANOVA. The data is expressed as Mean ± SEM. P<0.05 Group IV Ethanolic extract of M. fragrans was considered to be statistically significant. (EEMF400) seed 400 mg/kg p.o Group V Methanol fraction of M. fragrans RESULTS ((MFMF200) seed 200 mg/kg p.o. Group VI Methanol fraction of M. fragrans Effect of ethanolic extract of M. fragrans seed on (MFMF400) seed 400 mg/kg p.o. reaction time in rats on Eddy’s hot plate Group VII Benzene fraction of M.
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