Garcinia Kola Mitigates 3, 4
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Curr Neurobiol 2020; 11(2): 37-47 ISSN 0975-9042 Garcinia kola Mitigates 3, 4-methylenedioxymethamphetamine- induced Derangement in Cellular and Molecular Characterization in the Cerebellum of Wistar rat Owolabi Joshua Oladele, Okoro Iheanyichukwu, Adelodun Stephen Taiye, Olatunji Sunday Yinka, Olanrewaju John Afeez, Fabiyi Oluseyi Department of Anatomy, Ben Carson School of Medicine, Babcock University, Ilisan Remo, Ogun State Nigeria Abstract Background: 3, 4-methylenedioxymethamphetamine (MDMA) causes neurological disorders. Garcinia kola (GK) has neuroprotective property. We investigated the neuroprotective roles of GK on MDMA-induced cerebellar damage. Method: Sixty male Wistar rats (120 - 160g), 5 months old were grouped into six (n =10). Rats were fed with normal chow and water ad libitum. Group A was given distilled water, B and C received 100mg/kg and 200mg/kg GK respectively while D received 20mg/kg MDMA. Groups E and F received 100mg/kg and 200mg/kg GK respectively (pre-treatment), followed by intermittent 20mg/kg MDMA administration. All administrations were via oral route for 21days. Food, water intake, body temperature and weight were recorded. Rats were sacrificed and the brains were excised and used for assays. One-way ANOVA was used to analyze data followed by Student Neuman-Keauls test. Graph pad Prism 5 was used for analysis, p-values ≤ 0.05 were considered statistically significant. Results: Purkinje cells reduced in number and glial GFAP expression was increased. Serotonin, dopamine and glutamate levels showed no differences. MDMA treatment increased cytochrome c oxidase and glucose-6-phosphate dehydrogenase levels. Distance travelled was reduced while number of turns and foot slips were increased (p < 0.05). Weight loss and increase relative brain weight were observed (p < 0.05). It also increased the water intake and body temperature significantly. Conclusion: GK protected the cerebellum from MDMA toxicity in a dose-dependent manner through a conglomeration of evidences which include: Purkinje cells preservation, normalizing erratic gaits and acting as anti-diuretics. GK does not possess thermoregulatory function. Keywords: cerebellar damage, garcinia kola, MDMA, neurotoxicity, stereotypic movement, wistar rats. Introduction which is manifested as muscular tension, jaw clenching, tooth grinding (bruxism) and constant restless movement of the legs MDMA, otherwise known as Ecstasy, Adam, Lover’s speed, [8-10]. The increased muscle activity, together with a direct Superman etc. is one of the most commonly abused synthetic action of the drug on the thermoregulatory system in the brain drugs presently. The prevalence of its use in the recent time [11], leads to hyperthermia. Stiffness and pain in the lower-back among university students has been reported to be as high as and limb muscles are very common complaints during the first 13 to 39% in the U.S and United Kingdom [1-2]. Before now, 2-3 days after the use of MDMA. Users of MDMA from the mortalities and major adverse events due to recreational use of onset take the drug in pills or powder forms but there is now MDMA alone were uncommon and rarely reported [3]. However, a shift from those forms to the crystalline form due to its high the number of death and serious adverse effects of MDMA on purity. For example, the Ecstasy and Related Drugs Reporting users is on the increase in the recent time [4-5]. It has been System (EDRS) found that about 60 per cent of the users took postulated that life-threatening serotonin [5-hydroxytryptamine ecstasy in a high-purity crystal form. And they hypothesized (5-HT)] syndrome might contribute to mortalities and major those crystal MDMA users present riskier patterns of drug use adverse outcomes [6]. Ecstasy exposure has adverse effects on and lower health outcomes compared to non-crystal users. This new shift is the main reason for our choice of crystal forms of many physical functions even when taken in moderate doses for ecstasy for current this investigation [12]. the recreational purposes just as amphetamines do [7]. Because the basic action of the amphetamines involves increased arousal Cerebellum helps in the control of movement especially the and alertness, it is usually accompanied by an increase in tension, limbs. Although this structure is not necessary for the initiation 37 Curr Neurobiol 2020 Volume 11 Issue 2 Garcinia kola Mitigates 3, 4-methylenedioxymethamphetamine-induced Derangement in Cellular and Molecular Characterization in the Cerebellum of Wistar rat of motion, movements become erratic and distorted during known weight (250 g) of the powder was extracted in 1000 ml cerebellar damage due to toxicity insults on the movement of ethanol for 72 hours at room temperature. The extract was circuit [13]. Neurodegenerative diseases such as Alzheimer’s, filtered with Whatman No. 1 filter paper (Maidstone, UK) and Parkinson’s, Huntington’s, stroke and others are usually a the resulting filtrate concentrated in a Rotary Evaporator. The consequence of excess production of oxygen radicals from mixture was further transferred into steam bath where it was oxidative stress [14]. MDMA is known to induce neurotoxicity evaporated to give the required brownish-black residue. The in animal models, by the production of reactive species thus residue was stored in a desiccator until used. resulting in oxidative stress [15-16]. Chronic toxicity of MDMA Phytochemical screening: Basic phytochemical screening involves principally the brain, revealed through cerebral to detect the presence of alkaloids, saponins, tannins, flavonoids, imaging from previous studies [17-18] with consequences in steroids, anthraquinones, cardiac glycosides and carbohydrates memory loss especially short-term memory [19]. was carried out by adopting previous methods [27-31] (Table 1). Garcinia kola (GK) is a medium sized tree, ubiquitously Acute toxicity study (LD50) : The method described by found in West and Central African countries including Lorke [32] was adopted to determine the lethal dose (LD50). Nigeria, Ghana, Cameroon and Sierra Leone among others [20]. Almost every part of the plant contains phytochemicals Animal care and management that are reputed for their medicinal importance [21]. Among 5 months old sixty (60) male Wistar rats weighing between the phytochemicals found in GK seeds are tannins, saponins, 120 - 160 g were bred in the animal house of Babcock University alkaloids, cardiac glycosides and flavonoids (kolaflavone and Ilisan-Remo Ogun State, and used for this research. The rats 2 hydroxyflavonoids) [22]. Studies have shown that seeds of were grouped into six (A-F) n = 10. Rats were kept in a well GK possess anti-hepatotoxic, antioxidant [23], hypoglycaemic, ventilated standard housing conditions (temperature: 28-31°C; aphrodisiac, anti-cancer, anti-histamine, and antimicrobial photoperiod: 12 hours; humidity: 50-55%) in clean plastic cages properties among others [24]. Kolaviron (isolate of GK) has to acclimatise for one week. Ethical approval was obtained from specially been reported to elicit strong antioxidant activity, the Health Research Ethics Committee, Babcock University for in both in vivo and in vitro experimental models as seen in this research with a reference number (BUHREC-716/19). Rats its neuronal protection role against toxic substances [25]. A in all groups were fed with normal laboratory chow (Premier recent study has shown kolaviron to protect the neurons against Feed Mill Co. Ltd., Ibadan, Nigeria) and had access to water gamma radiation-induced oxidative stress in the brain of rats ad libitum. Group A served as control which was given distilled [26]. However, there is sparse report of the neuroprotective water. Groups B and C were given 100mg/kg GK and 200mg/ effect of GK on the cerebellum of Wistar rat against crystal kg GK respectively while group D received 20mg/kg of MDMA form of MDMA-induced neurotoxicity, hence, this study. This only. Groups E and F were also pre-treated with 100mg/kg GK research looked solely into the effect of crystal form of ecstasy and 200mg/kg GK respectively followed by 20mg/kg MDMA (at a dose of 20mg/kg given intermittently for the period of 21 administration every other day. All administrations were done days) on the cerebellum and the role of ethanolic extract of GK. through oral route for the period of 21days. The body weight, water-intake and food consumption of the rats were measured Materials and method throughout the period of treatment. Chemicals procurement MDMA stock preparation Crystal MDMA, ethanol (100%) as well as rabbit polyclonal 1g of crystalline MDMA was measured using sensitive anti-glial fibrillary acidic protein (GFAP) were purchased weighing balance (METTLER TOLEDO, China) in a cool room from Sigma Aldrich (St. Louis, MO, USA). Eosin 1% aqueous and dissolved in 100 ml of distilled water. This was kept inside was purchased from Biostain (Traralgon, Australia) while the fridge before use and the dosage for each rat was calculated Harris Hematoxylin was purchased from Harris Surgipath and administered using oral cannula. (Richmond, IL, USA) and Histofluid from Marien- feld (Lauda- Königshofen, Germany). Other chemicals were purchased from Table 1. Chemical compositions of GK. Preliminary ethanol screening of GK Sigma Aldrich excepted otherwise stated. reveals the following phytochemical constituents. Extraction of GK Phytochemical constituents Inference Alkaloids +++ GK seeds were procured from a local market at Ile-Ife, Flavonoids +++ Osun State,