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Issn: 2277–4998 IJBPAS, April, 2020, 9(4): 894-905 ISSN: 2277–4998 PHARMACOLOGICAL STUDIES ON ETHANOLIC EXTRACT OF COCCULUS PENDULUS COLLECTED FROM JHAL MAGSI DISTRICT, BALOCHISTAN, PAKISTAN KHAN NS1,2, MUHAMMAD S1*, JABBAR A1, AHMAD T2, HUSSAIN A2, ISHAQUE MMR2, ARSLAN M1 AND BALOCH S1 1: Department of Pharmacognosy, Faculty of Pharmacy & Health Sciences, University of Balochistan, Quetta, Pakistan 2: Department of Eastern Medicine, Faculty of Pharmacy & Health Sciences, City Campus, University of Balochistan, Quetta, Pakistan * Correspondence author: [email protected] th th th st Received 28 Oct. 2019; Revised 20 Nov. 2019; Accepted 27 Dec. 2019; Available online 1 April 2020 https://doi.org/10.31032/IJBPAS/2020/9.4.4848 ABSTRACT Balochistan is the largest province of the country and native home of many medicinal plants. Cocculus pendulus (Family: Menispermaceae) is a medicinal plant used in the crude form by the local people for digestive,pain reliving and depressive disorders. Current study was carried out to ethanolic extract of whole plant to determine phytochemical tests, acute toxicity, analgesic activity, forced swimming test and neuropharmacological activities. Ethanolic extract of C. pendulus showed presence of saponins, tannins and flavonoids in phytochemical tests, Inanalgesic activity significant (p<0.05) results in writhing test, tail immersion test, formalin test were observed. Ethanolic extract of C. pendulus also showed significant (p<0.05) CNS depressant results in force swimming test and neuropharmacological activities. Keywords: Analgesic, JhalMagsi, Cocculuspendulus 894 IJBPAS, April, 2020, 9(4) Khan NS et al Research Article INTRODUCTION Now a days the use of natural medicinal condition centrally acting muscle relaxant are product and supplements as accelerated use. Although these drug result in un wanted surprisingly over the previous three decade effect so therefore searching or trying to find which shows its enormous potential and an effective alternative as always being largely indicates the possibility of treating essential in this regard [3]. the disease [1]. There are a variety of herbal preparations Nervous disorders are becoming major recommended for pain relieving effect disorder and according to the Statistical data (analgesic) and reduces the anxiety (anti- 20- 40% of the general population, depressive drug). In search of new pain comprising more than 7% group of adult relieving effects and anti-depressive agent population, A prevalence figure show that from a large number of medicinally 14% of the population is suffering from significant plant, numerous research work nervous disease. There is another serious has been carried out, around the globe [4]. health problem of insomnia throughout the Present study was carried out to find out the both geriatric and teenage population, the neuropharmacological and analgesic recent information established the prevalence activities of the plant based on traditional associated with insomnia is equivalent to claim. major psychiatric disorder such as Cocculus pendulus depression. 88% of teenagers are suffering The genus Cocculus belongs to the family from chronic insomnia [2]. Menispermaceae. It comprises about 35 Skeletal muscle relaxant act peripherally at species, it occurs in Pakistan, India, Iran, the neuromuscular junction to reduce the Iraq, Saudi Arab. Its Anti-inflammatory, muscle tone. By acting on cerebrospinal axis wound healing and anti oxidant activities has neuromuscular blockers minimize muscle been reported [5]. firmness or initiate paralysis (cause MATERIAL AND METHOD dysfunction). For the purpose of surgery Plant specimen neuromuscular blocker are given which Plant was collected from Jhal Magsi District reduce the muscle tone and provide general of Balochistan, Pakistan, identified and aesthetic effect. Meanwhile for the pain full voucher specimen No NG-143 were muscle spasm and spastic neurological 895 IJBPAS, April, 2020, 9(4) Khan NS et al Research Article deposited at department of Pharmacognosy, mortality. The weight and possible University of Balochistan, Quetta. neurological lethality was observed[6]. Extraction Procedure Analgesic activity C. pendulus whole plant was macerated in Writhing test with acetic acid ethanol for 15 days, at reduce pressure the The pain releasingaction of C. Pendulus solvent was evaporated in rotary evaporator, ethanolic extract were anticipated by utilizing and dark green color semisolid residue was acetic acid writhing model in mice. The obtained. animal were grouped into four groups i.e. Test Animals Group I (control group), Group II & III Albino Mice weigh upalmost 25- 28 g were (ethanolic extract of C. pendulus 250 and utilized and housed in animal house of 500mg/kg treated group), Group VI (standard Faculty of Pharmacy and Health Sciences drug treatment). The 0.7% acetic acid according to standard protocol. administered intra peritoneal after 30 min of Preliminary Qualitative Phytochemical orally administration of test samples, Screening. standard drug and control vehicle. The C. pendulus ethanolic extract of was writhing (constriction of abdomen, and subjected to confirm the presence of some extension of hind legs) was observed for important phytoorganic compounds such half-hour [7]. asflavonoids, alkaloids, polyphenols, Tail Immersion Test saponins, anthraquinones, tannins, terpenes In this test,mice were separated into 4 groups and sterols were determined by using (5mice in each group). Control group (treated standard protocols [6]. with distil water1ml/kg), Group II & III Acute toxicity study (ethanolic extract of C. pendulus 250 and According to standard guideline (as per 500mg/kg treated group), Group VI (group of OECD 423 guidelines) the procedure was standard drug treatment). performed (OECD/OCDE. 2002). The oral After administration of each treatment, dose of ethanolic extract of C.pendulus from (about 2-3cm) tail of the each mice was 50 to 2000 mg/kg body mass was immersed into a water bath (temperature administered. Animal were held under maintained at 50 ± 1°C) and the time observation for 14 days to enlist conceivable consumed for the mice to withdraw its tail from the warm water or flick it known as the 896 IJBPAS, April, 2020, 9(4) Khan NS et al Research Article pain reaction time (PRT) was recorded for all maintained at 25 ± 1°C) 20cm depth. Mice the mice. 15 seconds were cut off time [8]. were placed for 6 minutes. Mice were Formalin test observed immobile when remain suspended In the right hind paw of the mice a formalin without struggling and generated only solution (2.5 % in 0.9 % sterile saline; 20 slight movements required to uphold the μl/paw sub plantar) was injected. head above the water. Decrease in mobility Observation was the period of time spent on time reflects the CNS depressant activity licking and biting the injected paw which is [10]. an indication of pain. After the 5 minutes of Neuropharmacological activites formalin injection the first phase (neurogenic Open Field Activity Test (OFT) pain) starts, than after 15–30 min of formalin The open field apparatus was consist of white injection second phase (inflammatory Plexiglas and measured 72 × 72 cm with response) starts formalin injection were given 36 cm walls. Red lines were drawn on the 30 minutes after to oral treatment (n = 5 per floor with a marker and were clearly visible group) with saline (control.), ethanolic through the clear floor. Mice were extract of C. pendulus (250 and 500mg/kg) administered (orally) saline 2ml/kg for and Diclofenic sodium (50 mg/kg) treated control group, with C. pendulus ethanolic groups [9]. extract (250 and 500 mg/kg) and Diazepam Forced swimming test 2mg/kg. The test was performed 30 min after In this test mice were randomly selected and the administration of the test substance. The divided into 4 groups with five mice each. mice were placed in the open field box for Saline 2ml/kg for control group, along with 10 minutes, and their square crossed by the C. Pendulus ethanolic extract (250 and 500 mice were recorded [11]. mg/kg) and Diazepam 2mg/kg administered Hole-board test orally. The Mice are placed in an This test is mainly used for measuring inevitableclearchamber that is filled with exploratory behaviours. This apparatus is water and their escape related comprises of a wooden, grey box, measuring movementbehaviour is measured, mice were (40 cm × 40 cm). The walls were 25 cm high, individually put in a round bottom tank with sixteen equidistant holes (3 cm in (which was about 46 cm tall × 20 cm in diameter), the control group were breadth) filled up with tap water (temperature administered C. Pendulus ethanolic extract 897 IJBPAS, April, 2020, 9(4) Khan NS et al Research Article (250 and 500 mg/kg) and Diazepam 2mg/kg in the beaker and number of upwards treated group. The observation was to count movements were observed for 10 minutes. the all number of crosses through the hole Statistical Analysis from one chamber for 10 minutes [12]. Means ± standard deviation (SD) were used Stationary Rod Test for expression of data. Level of significance The Stationary Rod Test one of the most between the means of treated groups and commonly used tools for testing motor control group, was determined by one-way coordination, balance and learning ability. ANOVA. Differences were considered The test apparatus consisted of horizontal statistically significant at p< 0.05 [6]. stainless steel rods with two platforms at the RESULTS ends. The mice first trained to walk on the Phytochemical
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