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Maerua Angolensis Extract Reduces Allodynia and Hyperalgesia in a Mouse Model of Vincristine-Induced Peripheral Neuropathy

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Maerua Angolensis Extract Reduces Allodynia and Hyperalgesia in a Mouse Model of Vincristine-Induced Peripheral Neuropathy Journal of Applied Pharmaceutical Science Vol. 6 (05), pp. 124-130, May, 2016 Available online at http://www.japsonline.com DOI: 10.7324/JAPS.2016.60519 ISSN 2231-3354 Maerua angolensis Extract Reduces Allodynia and Hyperalgesia in a Mouse Model of Vincristine-Induced Peripheral Neuropathy Hosea Azi Iliya1,2, Wonder Kofi Mensah Abotsi2, Charles Benneh2, Eric Woode2* 1Department of Pharmacology, University of Jos, Jos, Nigeria. 2Department of Pharmacology, Faculty of Pharmacy and Pharmaceutical Sciences, College of Health Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana. ABSTRACT ARTICLE INFO Article history: Allodynia and hyperalgesia comprise the main and frequent symptoms suffered by patients with neuropathic Received on: 30/12/2015 pain, which responds poorly to therapy. An earlier study reported that stem bark extracts of Maerua angolensis Revised on: 15/01/2016 exhibited dose-dependent anti-nociceptive effect against the neurogenic and inflammatory phases of the Accepted on: 20/02/2016 formalin test. The current study evaluated the effect of petroleum ether/ethyl acetate stem bark extract of Available online: 28/05/2016 Maerua angolensis on vincristine-induced neuropathy. Neuropathic pain was induced by intraperitoneal injection of vincristine (0.1 mg/kg/day) using a 5-day-on, 2-day-off schedule over 12 days. On day 15, baseline Key words: responses were measured in the Randall-Selitto mechanical hyperalgesia test and paw withdrawal tests (using Maerua angolensis, Von Frey filaments and cold water at 4.5 °C) and mice that developed allodynia/hyperalgesia were randomly Neuropathic pain, Von Frey assigned into 7 groups. Normal saline (i.p.), pregabalin (10, 30, and 100 mg/kg, p.o.) and extract (3, 10, and 20 filaments, allodynia, mg/kg, p.o.) were administered to the individual groups. Allodynia/hyperalgesia was measured hourly for 5 hyperalgesia. hours post treatment. The extract produced significant (P<0.05) and dose-dependent inhibition of vincristine- induced mechanical hyperalgesia, tactile and cold allodynia responses. In all, the study shows that oral administration of Maerua angolensis stem bark extract inhibits vincristine-induced neuropathy in mice suggesting that it may exert analgesic effect in cancer patients with vincristine-induced neuropathic pain. INTRODUCTION Thus, the search for new chemical entities that can act as promising molecules to treat chemotherapy-induced neuropathic Neuropathic pain is a common problem that presents a pain has emerged. Medicinal plants are potential sources of major challenge to health-care providers owing to its complex commercial drugs and lead compounds in drug development natural history, uncertain aetiology and poor response towards (Zhang, 2004) forming important sources of new chemical therapy (Quintans et al., 2014). It is a chronic pain condition that substances with potential therapeutic effects (Ebadi, 2007). arises from a disease or injury to the central nervous system Maerua angolensis DC (Family: Capparidaceae) is a medicinal (CNS) or the peripheral nervous system (PNS) leading to its plant used traditionally to relieve pain (Mothana et al., 2009; Meda damage or abnormal function. Common symptoms of neuropathic et al., 2013). pain include sensory abnormalities such as burning sensations, The petroleum ether/ethyl acetate stem bark extract of hyperalgesia, allodynia, hyperesthesia and dysesthesia (Schim, this plant has been demonstrated to possess anti-nociceptive effect 2009). The current analgesics are unable to treat cancer in chemical, mechanical and thermal behavioural models of chemotherapy-induced neuropathic pain which is severe enough nociception in rodents (Iliya et al., 2014b). In another study (Iliya for patients to discontinue their cancer chemotherapy treatment et al., 2014a), the authors speculated that the extracts could be and worsens the quality of their life (Lynch et al., 2005; Wolf et effective against neuropathic pain since the second phase of the al., 2008; Park et al., 2012). formalin test is predictive of activity in neuropathic pain models (Alessandri-Haber et al., 2004; Vissers et al., 2006). As a follow- * Corresponding Author up to confirm that assertion, the current study investigated the anti- Eric Woode, Department of Pharmacology, Faculty of Pharmacy and nociceptive effect of the petroleum ether/ethyl acetate stem bark Pharmaceutical Sciences, College of Health Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana extract of Maerua angolensis in the vincristine-induced Email: [email protected] neuropathic pain model. © 2016 Hosea Azi Iliya et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial- ShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/). Iliya et al. / Journal of Applied Pharmaceutical Science 6 (05); 2016: 124-130 125 MATERIALS AND METHODS following behavioural testing for mechanical, tactile and cold allodynia and/or hyperalgesia, the mice were administered Plant materials vincristine (0.1 mg/kg/day, i.p.) or normal saline (10 ml/kg/day, Fresh stem barks of Maerua angolensis were collected in i.p) in two cycles of five consecutive days with two days off August 2012 from the Samaru campus of Ahmadu Bello between the cycles (days 1-5 and days 8-12) to induce neuropathic University, Zaria, Nigeria and were identified by Dr. Kofi Annan pain On day 15, baseline responses were measured in the Von Frey of the Department of Herbal Medicine, Faculty of Pharmacy and filaments (4, 8 and 15 g), Randall-Selitto mechanical hyperalgesia Pharmaceutical Sciences, Kwame Nkrumah University of Science and cold allodynia (cold water at 4.5 °C) tests. Mice that and Technology (KNUST), Kumasi, Ghana. A voucher specimen developed allodynia and/or hyperalgesia were randomly assigned (KNUST/FP/12/051) was kept at the herbarium of the Faculty. into 7 groups and subsequently treated with MAE (3, 10, and 20 mg/kg, p.o.), pregabalin (10, 30, and 100 mg/kg, p.o.) or normal Extraction saline (10 ml/kg, i.p.). Four kilograms of the dried powdered stem bark was Allodynia and/or hyperalgesia was measured at hourly sequentially extracted with petroleum ether (10 L) and ethyl intervals for 5 hours post-treatment using three sets of experiments acetate (10 L) over a 4-day period by cold maceration. The liquid namely Von Frey (tactile allodynia, intermediate and mechanical extracts were filtered and each concentrated separately in a rotary hyperalgesia) test, Randall-Selitto mechanical hyperalgesia test evaporator under reduced temperature and pressure to give and cold allodynia (cold water at 4.5 °C) test to assess the effect of yellowish syrupy mass. MAE on vincristine-induced neuropathic pain. The extracts were further dried in an oven at 50 °C to obtain yields of 2.23 % (w/w) (petroleum ether) and 3.87% (w/w) Assessment of tactile allodynia, intermediate and mechanical (ethyl acetate). Since thin-layer chromatography (TLC) analysis of hyperalgesia the two extracts revealed similar composition of phytochemical Tactile allodynia was evaluated by means of Von Frey constituents, the petroleum ether and the ethyl acetate extracts filaments (IITC Life Science Inc. Model 2888, Woodland Hills, were combined. The combination is subsequently referred to as CA, USA) with bending forces of 4 g. Intermediate and petroleum ether/ethyl acetate stem bark extract of Maerua mechanical hyperalgesia were evaluated with Von Frey filaments angolensis or MAE in this study. of bending forces of 8 and 15 g respectively. Responses to 15 g are best explained as hyperalgesia (exaggerated pain response from a Animals normally noxious stimulus) as normal mice withdraw from this Male Swiss albino mice (20-25 g) were used after stimulus 5-10 % of the time; while the responses to 8 g are approval of protocol for the study by the local ethical committee intermediate. for animal handling and experimental procedure. All animals were In ascending order of force, each filament was applied to housed in groups of five in stainless steel cages (34 × 47 × 18 cm) the mid-plantar area (dodging the base of the tori) of each hind with softwood shavings as bedding in the animal facility of the paw five times, with each application held for 5 s. Withdrawal Department of Pharmacology, KNUST, with free access to food responses to the Von Frey filaments from both hind paws were and water and were maintained under normal laboratory conditions counted and expressed as an overall percentage (Flatters et al., of humidity, temperature (25 ± 1 °C) and a 12 h/12 h day/night 2004; Siau et al., 2006). cycle. Each animal was used only once. The investigation conforms to the Guide for the Care and Use of Laboratory Animal Assessment of mechanical hyperalgesia in the Randall-Selitto published by the US National Institutes of Health (NIH No. 85-23, revised 1996). In all the experimental studies each group consisted test of 5 animals. Mechanical nociception was determined with the IITC Life Science Model 2888 (Woodland Hills, CA, USA). The mouse’s hind paw was placed into the pressure applicator, and a Drugs and chemicals Pregabalin (Pfizer Pharmaceuticals, Arzneimittelwerk constantly increasing pressure stimulus (maximum cut-off of 250 Godecke, Freiburg, Germany) and vincristine sulphate (Celon g) was applied to the dorsal surface of the paw until withdrawal or Laboratory Ltd, Gajularamaram, India) were used. vocalization happened, at which weight the nociceptive threshold value was documented. For each mouse, two readings were Phytochemical screening documented for each hind paw, and the results reported as Preliminary phytochemical tests were performed on the the mean value of readings from both hind paws (Woode et al., extract using methods described by Trease and Evans (1989). 2013). Vincristine-induced neuropathic pain Assessment of cold allodynia Vincristine sulphate was dissolved in normal saline and Cold allodynia was evaluated by immersion of the stored as a stock concentration of 1 mg/10 ml at 4 °C. Immediately animal’s hind paw into a water bath containing cold water (4.5 °C) 126 Iliya et al.
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