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Parklane Enugu State. Nigeria December 30, 2018 Archives • 2018 • vol.3 • 98-104 ANXIOGENIC AND NEURODEGENERATIVE EFFECTS OF AKAKI EXTRACT ON THE CEREBELLUM OF ADULT WISTAR RATS. Elizabeth Finbarrs-Bello1*; Oluwatomilayo Patience Ojo2; Egwu Eni Ogbonnia3 1 Enugu State University of Science and Technology, [Department of Anatomy, Faculty of Basic Medical Sciences] Parklane Enugu State. Nigeria. 2 University of Ilorin, [Department of Anatomy, Faculty of Basic Medical Sciences], Ilorin, Nigeria 3Ebonyi State University, [Department of Anatomy, Faculty of Basic Medical Sciences] Abakaliki, Ebonyi State, Nigeria. [email protected] Abstract The combination of Costus afer, Sarcocephalus Latifolius and Culcasia Scandens (akaki extract) is used as treatment for various mental disorders in some communities in South-East Nigeria. This study was designed to investigate the effect of akaki extract on the anxiety and histology of the cerebellum of Wistar rats. Phytochemical analysis for the presences of bioactive agents was carried out using standard chemical methods. Twenty rats of average weight 200grams were randomized into four groups (n=5); control group, received feed and water, while the rats in the experimental groups were treated daily with 3mg/kg, 6mg/kg and 9mg/kg of the akaki extract for seven days. On day 8, anxiety text was carried out using the elevated plus maze (EPM) apparatus while cerebellar histoarchtecture was demonstrated by routine haematoxylin and eosin method. Phytochemical analysis revealed presences of saponins, phenols, flavonoids, tannins and glycosides. The extract exhibited anxiogenic effect in the EPM and showed dose dependent neurodegenerative changes (atrophy of neurons) in the experimental groups compared to the control. In conclusion, akaki extract induces neurotoxic and anxiogenic effects on the cerebellum. Keywords: Akaki Extract, Phytochemicals, Neurotoxic, Anxiogenic, Cerebellum http://pharmacologyonline.silae.it ISSN: 1827-8620 PhOL Elizabeth, et al. 99 (pag 98-104) Introduction African peach, Guinea peach, Negro peach [11]. In Anxiety disorder is characterized by a variety of Nigeria, it has various ethnic names; Egbesi in neuroendocrine, neurotransmitter, and Yoruba and Ulefum, Urevum, Ubulu-inu or Obarailu neuroanatomical disruptions [1]. The in Igbo [12]. It also has a wide range of medicinal neurotransmitters in the brain constitute excitatory application in the management of epilepsy and and inhibitory neuronal networks that maintain a depression [13]. It also has anticonvulsant, anxiolytic finely tuned balance of activity critical for normal and sedative properties [14]. functioning. These include the glutamate and The Culcasia scandens belongs to the order gamma-aminobutyric acid (GABA) [2]. In the healthy Arales, family Araceae and sub-family brain, gamma-aminobutyric acid (GABA) is regulated Philodendroideae of the genus Culcasia [15]. It is an by neurons and glia located in different areas of the epiphytic climbing herb with slender wiry stems, of brain such as the hippocampus, amygdala and the fringing forest and savanna in West Africa [7, cerebellum. In the cerebellum formed the 15]. The anti-rheumatic, anti-emetic and anti- GABAnergic neuron and their receptors which are abortifacient actions as well as possessing analgesic heterogeneously distributed in cerebellar cortex. properties of C. scandens have been documented Increases in anxiety are accompanied by GABAergic [16, 17]. system deregulation which has a large impact on the brain [3]. Notably, to date no available study has In Nkanu community of Enugu State, South-east examined the impact of anxiety on the cerebellum Nigeria, the decoction of the three herbs (Costus in brain diseases characterized by GABA afer, Sarcocephalus latifolius and Culcasia scandens) dysregulation including anxiety disorders. known as Akaki in this study is used as treatment for various mental disorders. Our previous studies Studies have discovered anxigenic and anxiolytic reported adverse effect of the three herbal properties of many interesting properties of plants combinations on the frontal and temporal lobes of based on the knowledge of their traditional the cerebral cortices while investigating the applicability in the treatment of certain disease behavioural effects as well [8,9]. We opined in the condition [4, 5]. The massive use of herbs in the present study that the alterations in GABAergic treatment of diseases is encouraged by their neurons which played a major role in changing the efficiency, their availability and low cost [6]. Akaki is neuronal activity observed in anxiety in the areas of a combination of three medicinal plants Costus afer, the cerebral cortex would likely involve the Sarcocephalus latifolius and Culcasia scandens used cerebellum in the holistic view. Thus providing a in folklore medicine in the management of central better understanding of the impact of anxiety on nervous system disorders. cerebellum and how it may contribute to anxiety Costus afer is a genus of perennial tropical from this scientific perspective. Hence, the study herbaceous plant in the family of Costaceae. Its investigated the phytochemical constituents, common names are: ginger lily, gingembre spiral or behavioural effect on anxiety and histology of the bush cane. In Nigeria it has various ethnic names; cerebellum Igbos calls it Opote, Okpoto or Okpeteohia; Methods Ukhueroha in Edo, Mbritem in Efik and Ibibio, Kakii- zuwaa in Hausa, Achikku in Tiv and Atare-egun in Plant material and extract preparation: Yoruba. It is often found in the forest belt The three plants (Costusafar, Sarcocephalus throughout tropical Africa [7-9]. The hot water latifolius and Culcasia scanden) were collected extract of its leaves and stem is widely used from Agbani village, Nkanu, Enugu State, Nigeria traditionally to treat minor epileptic attacks and (06°19′ 0″North; 07° 33′ 0″ East) during the month of insomnia [10]. July, 2017. The plants were authenticated at the Sarcocephalus latifolius belongs to genus of Department of Plant Science and Biotechnology, flowering plant in the Rubiaceae family, a shrubs or University of Nigeria Nsukka. The fresh leaves of tree native of tropical Africa. Common names are Costus afer, Sarcocephalus latifolus and Culcasia scandens were washed thoroughly with distilled http://pharmacologyonline.silae.it ISSN: 1827-8620 PhOL Elizabeth, et al. 100 (pag 98-104) water and room-dried for seven days and Test for phenols: a small portion (1ml) of the pulverised into fine powder. Two hundred (200) extract was added to 1ml of water and few drops of grams of each plant power was measured and 5% NAOH were added. No orange colouration was mixed together, and then the powdered mixture observed in the test, indicating the absence of was divided into two portions. A portion was taken phenol. to Project Development Institute (PRODA) Enugu Test for saponins: the presences of saponins were for phytochemical analysis, while the rest portion detected using frothing test. In the test 2ml of the was used for the preparation of the extract. extract was vigorously shaken for 2minutes. Three hundred (300) grams of the powder was Frothing observed in the extract indicated the submerged into 1250 ml of hot water and allowed to presences of saponins. boil for 45 minutes as to mimic the traditional or Test for steroids: presence of steroids was indigenous herbal method. After which it was investigated using Salkowski test in which 5 drops of allowed to stand for 24 hours and filtered using concentration (H2SO4) were added to 1ml of the muslin cloth. The filtrate was then concentrated into extracts. No red coloration was observed in the a green syrupy mass under reduced pressure at 60°C mixture, indicating the absence of steroids. using a rotary evaporator. It was further dried in a hot air oven at 50°C for a week and kept in a Test for tannins: exactly 2 ml of the extract was refrigerator until further use. The yield was 30.3%. boiled gently for 2 minutes and allowed to cool. This crude extract is subsequently referred to as Three drops of ferric chloride solution were added. akaki extract in this study. No green colouration was observed, which indicated the absence of tannins. Phytochemical screening: Experimental animals: The presence of saponins, alkaloids, flavonoids, steroids, phenols, glycosides, and tannins in the A total of twenty rats (Wistar rats ) 5 to 6 weeks combined leaves of Costus afer, Sarcocephalus old, and weighing 150g to 180g were obtained from latifolius and Culcasia scandens (Akaki) were tested Department of Biotechnology, Ebonyi State by their standard methods described by Elizabeth et University, Abakaliki. The rats were housed al. [1]. individually in a temperature-controlled (20 - 22°C) room under 12 h : h light/dark cycles and had ad Test for alkaloids: exactly one millilitre (1ml) of 1% libitum access to standard rodent chow food pellets Hydrochloric Acid (HCL) was added to 3ml of the and tap water for two weeks. After this habituation extract in a test tube. The mixture was heated for pe¬riod, at the age of 8 weeks, the rats were 20minutes in a water bath, allowed to cool, and randomized into 4 equal and weight-matched then filtered. Two drops of Wagner’s reagent were groups: a control group (controls, n = 5) and added to 1ml of the filtrate. A reddish brown experimental groups A, B and C (N = 5 in each precipitate observed indicated the presence of group) by generating random numbers using alkaloids. standard. All conditions and handling of the animals Test for flavonoids: 1ml of 10% sodium hydroxide were ap¬proved by the Ethics Committee of Faculty (NaOH) was added to 3ml of the extract. No yellow of Basic Medical Sciences, Ebonyi State University coloration was observed in the test indicating the Certificate Number: FBMEC 440 and conducted absence of flavonoids. according to the Declara¬tion of Helsinki Test for glycosides: 10ml of 50% tetraoxosulphate Experimental design: (VI) Acid (H2SO4) was added to 1 ml of the extract in The animals were divided into four groups (n=5); a test tube. The mixture was heated in boiling water the control group received 0.1 ml of saline, while the for 15minutes.
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