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Anxiolytic Activity of Aqueous Extract of Bridelia Micrantha (Hochst) Baill (Euphorbiaceae) in Mice

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Anxiolytic Activity of Aqueous Extract of Bridelia Micrantha (Hochst) Baill (Euphorbiaceae) in Mice WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES Minkoulou et al. World Journal of Pharmacy and Pharmaceutical Sciences SJIF Impact Factor 7.421 Volume 8, Issue 2, 01-18 Research Article ISSN 2278 – 4357 ANXIOLYTIC ACTIVITY OF AQUEOUS EXTRACT OF BRIDELIA MICRANTHA (HOCHST) BAILL (EUPHORBIACEAE) IN MICE EXPOSED TO CHRONIC IMMOBILIZATION STRESS Delphine Mireille Ze Minkoulou1*, Jean Pierre Omam Omam2, Antoine Kavaye Kandeda1, Florence Ngueguim Tsofack1, Elisabeth Ngo Bum3, Théophile Dimo1 1Department of Animal Biology and Physiology, Faculty of Science, University of Yaoundé I, P.O. Box 812 Yaoundé, Cameroon. 2Higher Teachers’ Training College, University of Yaoundé I, P.O. Box 47, Yaoundé, Cameroon. 3Department of Biological Sciences, Faculty of Science, University of Ngaoundéré, P.O. Box 454, Ngaoundéré, Cameroon. ABSTRACT Article Received on 21 Nov. 2018, Anxiety disorders are a major public health concerning worldwide. The Revised on 11 Dec. 2018, current anxiolytics are ineffectives and associated with major side Accepted on 02 Jan. 2019 DOI: 10.20959/wjpps20192-13021 effects. Thus, medicinal plants with better effects constitute source of new pharmaceutical drugs. This work aimed to assess the anxiolytic- like properties of aqueous bark extract of Bridelia micrantha on *Corresponding Author behavioral and neurochemical characteristics, using male mice Delphine Mireille Ze Minkoulou subjected to chronic immobilization stress. The anxiolytic properties of Department of Animal B. micrantha were assessed in mice after repeated stress by Biology and Physiology, immobilization. Mice received distilled water (10 ml/kg, p.o), Faculty of Science, diazepam (2 mg/kg, i.p) or B. micrantha extract (76, 152 or 305 mg/kg, University of Yaoundé I, p.o) one hour prior to daily exposure to stress by immobilization in the P.O. Box 812 Yaoundé, Cameroon. adapted plastic cone (3h/day) for 14 days. The behavioral parameters were evaluated using Elevated Plus Maze (EPM), Open Field (OF) and Hole-Board (HB) tests. The plasma levels of serotonin and corticosterone were determined by ELISA technique. Like diazepam, the extract increased (p0.001) the number of open arms entries and the time spent in open arms of the EPM. The extract elicited the increase of crossing and grooming number, and the time spent in centre of the of. On the HB, the extract increased (p< 0.001) the number of head dipping and decreased the number of www.wjpps.com Vol 8, Issue 2, 2019. 1 Minkoulou et al. World Journal of Pharmacy and Pharmaceutical Sciences crossing. The extract increased (p<0.001) plasma serotonin levels and decreased corticosterone levels. The chronic restraint stress-induced behavioral and neurotransmitters levels abnormalities were attenuated by the extract. These overall results suggest that aqueous bark extract of B. micrantha might have a promising therapeutic potential for stress-related disorders. KEYWORDS: Chronic Immobilization Stress, Anxiety, B. Micrantha, Serotonin, Corticosterone. INTRODUCTION Acute and chronic stresses are characterized by the physiological changes that occur in response to novel or threatening stimuli. Chronic stress has been linked to the pathophysiology of mood including anxiety disorders and depression.[1] Mental Stress is defined as the non-specific response of the body to any demand imposed upon it.[2] Stress is known to alter the physiological homeostasis of the organism and complex mechanisms contribute to the breakdown in adaptation processes resulting in various visceral, endocrinal, and behavioral changes.[3] The neuroendocrine damages in response to acute and chronic stresses are mediated by both the sympathetic nerve system and the hypothalamus-pituitary- adrenal (HPA) axis, which lead to the regulation of the release of dopamine, norepinephrine, serotonin, glutamate, and corticotropin-releasing and adrenocorticotropic hormones in central nervous system (CNS) and the secretion of glucocorticoids in plasma.[4] Cortisol and corticosterone are thus often used as biomarkers for stress, depressive and anxiety disorders.[5] Stress plays the main role in pathogenesis of mental disorders.[6,7] A host of chronic psychiatric disease states like melancholic depression, anorexia nervosa, panic disorders, anxiety disorders and cognitive dysfunction have been reported to involve abnormality of stress axis.[8] Restraint stress can induce a series of dysfunctions of central nervous system, such as cognitive impairment, anxiety, depression, amnesia, and insomnia. A number of chronic stress models, including electric footshock (EF) stress, forced swimming, noise stimulus and immobilization, have been employed to induce anxiety disorders in the past decades.[9] However, elevated plus-maze is a widely used test, which is based on the natural aversion of rodents to heights and open spaces, and which has been validated to be suitable for the assessment of anxiety disorders for both mice and rats.[10] As another characteristic behavior of the anxiety state of animals, the open field and hole-board tests are also used as a screening model for the detection of anxiolytics.[10,11] Anxiety disorders in particular, affect 1/8 th of total population worldwide and have become one of important www.wjpps.com Vol 8, Issue 2, 2019. 2 Minkoulou et al. World Journal of Pharmacy and Pharmaceutical Sciences research interest in psychopharmacology during this decade.[12] The hallmark of anxiety disorders is marked, persistent and excessive or unreasonable fear that is experienced to a degree that significantly interferes with everyday life.[13] Chronic stress and chronic stress- induced anxiety disorders have become increasingly more important public health concerns in recent years. However, the ranges of available pharmacotherapy like the benzodiazepines for the treatment of anxiety disorders induced by chronic stress are limited and suboptimal with regard to efficacy and tolerability.[14] A search for novel pharmacotherapy from medicinal plants for psychiatric illnesses has progressed significantly in the past decade. This is reflected in the large number of herbal preparations for which psychotherapeutic potential has been evaluated in a variety of animal models.[15] Several plants have proven anxiolytic- like effects in animal models.[16,17,18] Bridelia micrantha (Hochst.) Baill. is a small to medium sized tree belonging to the family Phyllanthaceae (formerly Euphorbiaceae), commonly known as mitzeerie or coastal golden leaf.[19] The following activities have been reported from B. micrantha: anthelmintic, antibacterial, anticonvulsant and sedative, antidiabetic, antidiarrhoeal, antifungial, anti-Helicobacter pylori, antimycobacterial, antinociceptive, antioxidant, antiplasmodial, antischistosomal, antiviral, hepatoprotective, insecticidal, β- lactamase inhibitor.[19] Thus it is not surprising that the bark, leaves and roots of B. micrantha are widely used as herbal medicines in tropical Africa.[19] The powdered bark is used for diseases of the central nervous system like epilepsy and insomnia in Cameroon.[19,20] Multiple classes of phytochemicals including alkaloids, anthocyanidin, anthraquinones, carbohydrates, cyanogenic glycoside, essential oil, ester, flavonoids, oxalate, phenolic compounds, saponins, sterols, tannins, terpenoids as well as several minerals have been isolated from the bark, fruits, leaves and roots of B. micrantha.[19,21] The aim of the present study was to evaluate the neuropharmacological activities of aqueous bark extract of B. micrantha, in an experimental model of chronic stress-induced anxiety disorders by investigating the behavioral parameters, using the elevated plus-maze, open field and hole-board tests, and determining its influences on the levels of serotonin and corticosterone in the plasma. MATERIAL AND METHODS Material Plant material: The barks of B. micrantha were collected in the immediate vicinity of Yaoundé (Nkombassi), Cameroon, during the dry season in July 2010.The plant materials were identified (voucher specimen Nº 9678/SRF/Cam) at the National Herbarium of Cameroon in Yaoundé. www.wjpps.com Vol 8, Issue 2, 2019. 3 Minkoulou et al. World Journal of Pharmacy and Pharmaceutical Sciences Animals Adult male Swiss albinos mice weighing 20-25g and about 2-3 months from our breeding stock (Animal house of the laboratory of Animal Physiology of the University of Yaoundé I) were used in this study. The animals were housed at 25±3°C with 12:12 h light and dark cycle. They had free access to food and water. The animals were acclimatized for a period of seven days before the study. All animal handling procedures were done in accordance with National Ethic Guidelines (FWA-IRB00001954), and the experiments designed to minimize the number of animals used and to minimize their suffering. Chemicals Diazepam was obtained from Roche (France). Serotonin and Corticosterone kits were purchased from IBL INTERNATIONAL GMBH (Hamburg, Germany). Methods Preparation of the aqueous extract The aqueous extract and doses of B. micrantha were obtained based on the traditional medicine protocol. The collected barks of the plant were dried under room temperature. The dried and powdered bark (100 g) of B. micrantha was macerated in 1l of distilled water for 1 hour. The mixture was boiled for 20 min. After cooling, the supernatant was collected and filtered using Whatman filter paper Nº1. In another experiment the filtrate was then evaporated to dryness using an oven at 45°C giving aqueous extract with a 6.1% yield. Experimental
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