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Possible Oligodendrogenic Effect of Omega 3 and Green Tea R.A

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Possible Oligodendrogenic Effect of Omega 3 and Green Tea R.A View metadata, citation and similar papers at core.ac.uk brought to you by CORE Foliaprovided Morphol. by Via Medica Journals Vol. 78, No. 3, pp. 564–574 DOI: 10.5603/FM.a2018.0105 O R I G I N A L A R T I C L E Copyright © 2019 Via Medica ISSN 0015–5659 journals.viamedica.pl Acrylamide-induced adverse cerebellar changes in rats: possible oligodendrogenic effect of omega 3 and green tea R.A. Imam, H.N. Gadallah Department of Anatomy and Embryology, Faculty of Medicine, Cairo University, Cairo, Egypt [Received: 13 September 2018; Accepted: 10 December 2018] Background: Humans are widely exposed to acrylamide (ACR) and its neurotoxicity is a significant public health issue attracting wide attention. The aim of the study was to investigate ACR-induced adverse cerebellar changes in rats and study the possible oligodendrogenic effect of omega 3 and green tea. Materials and methods: Twenty-four adult albino rats weighing 150–200 g were randomly divided into four equal groups (6 rats each): control group (Group I), the rats that received ACR 45 mg/kg/day (Group II), the rats that received ACR concomitant with omega 3 at a dosage of 200 mg/kg/day (Group III), the rats that received ACR concomitant with green tea dissolved in drinking water at a dosage of 5 g/L (Group IV). The rats were euthanized after 8 weeks of the experiment. Malondialdehyde (MDA) and glutathione (GSH) were measured in cerebellar homogenates. Sections of 5 µm thickness from specimens from the cerebellum were stained with haematoxylin and eosin, silver stain and immunohistochemical stains: platelet-derived growth factor alpha (PDGFα; for oligodendrocytes), glial fibrillary acidic protein (GFAP; for astrocytes) and BCL2 (antiapoptotic). Results: Omega 3 and green tea had improved MDA and GSH as compared to the ACR group. Histologically, the ACR group showed variable degrees of cellular degeneration. Omega 3 had induced oligodendrogenesis in Group III. The optical density of silver stain was significantly (p < 0.05) increased in Groups III and IV as compared to the ACR group. Area per cent of positive PDGFα was significantly increased in the ACR + omega 3 group as compared to the ACR group. Area per cent of positive GFAP was significantly decreased in Groups III and IV as compared to the ACR group. Area per cent of positive BCL2 was significantly increased in the omega 3-trated group as compared to the ACR group. Conclusions: Concomitant administration of omega 3 or green tea with ACR might mitigate the adverse cerebellar changes caused by ACR thanks to an oli- godendrogenic effect of omega 3. (Folia Morphol 2019; 78, 3: 564–574) Key words: acrylamide, cerebellum, omega 3, green tea, rats, oligodendrocytes Address for correspondence: Dr. R.A. Imam, Anatomy and Embryology Department, Faculty of Medicine, Cairo University, Cairo, Egypt, tel: 01006114696, e-mail: [email protected] 564 R.A. Imam, H.N. Gadallah, Acrylamide-induced adverse cerebellar changes in rats INTRODUCTION lead-treated rats, which resulted from its inhibitory Acrylamide (ACR) is an industrial neurotoxic chem- effect on free radical chain reactions generated during ical that has been found in carbohydrate-rich foods oxidative stress caused by lead and from an increase cooked at high temperatures such as potato chips in antioxidant enzyme capacity [23]. Over the last two [15]. Therefore, humans are widely exposed to ACR decades polyphenols, one of green tea constituents, and its neurotoxicity in humans is a significant public have drawn attention as promising natural dietary health issue attracting wide attention [19]. It was molecules for the prevention of ageing and neuro- also associated with carcinogenicity and reproductive degenerative diseases [22]. Recently, dietary polyphe- toxicity [6]. The neurotoxicity of ACR has been known nols, endowed of antioxidant and anti-inflammatory to affect nerve terminal and cysteine residues on the properties, had been reported to extend brain health functionally important presynaptic proteins, resulting span as they were actively investigated as potential in inhibition of neurotransmitter release and eventual adjuvants to support proliferation and survival of process degeneration [25, 26]. Cerebellum controls neural progenitors, and counteract age-dependent the maintenance of equilibrium (balance), influenc- neurogenic decline [36]. es posture and muscle tone as well as coordinating The present study deals with the influence of movement [8]. Cerebellar ataxia may be acquired from green tea extract and omega 3 fatty acids in ACR-in- exposure to many toxic materials [39]. Myelin is an duced adverse cerebellar changes in rats. electrical insulator that increases conduction velocity of nerve fibres and is the physical basis for rapid saltatory MATERIALS AND METHODS conduction (in which impulses jump from one node Animals of Ranvier to another) [29]. Inflammatory damage to The experiment was ethically approved by the the oligodendroglia and white matter is involved in the Institutional Animal Care and Use Committee of Cai- pathogenesis of demyelinating diseases such as multi- ro University (CU-IACUC) under the number of CU/ ple sclerosis [5]. Oligodendrocyte is the cell responsible /III/F/48/18. The present study was carried out on for producing central nervous system (CNS) myelin by 24 male albino rats weighing 150–200 g. The animals the concentric layers of its plasma membrane [34]. It is were housed in cages, under standard laboratory located in both the grey and the white matter of the and environmental conditions with free access to CNS: interfascicular oligodendrocytes (white matter) food and water at a temperature of (20 ± 2ºC) with and satellite oligodendrocytes (grey matter only) [14]. a natural 12-h light/dark cycle and free access to The observation that Eskimos had a very low in- standard pellet chow and drinking water ad libitum. cidence of atherosclerosis and its complications, had Animals were obtained from Animal House, Faculty led to the first indication of a protective effect of of Medicine, Cairo University. The rats were set in the fish oil on atherosclerosis. Today, fish oil is one of laboratory for a period of 2 weeks for acclimatisation the most popular nutraceuticals available in health before carrying out the experiment. food stores [27]. Surprisingly, fish oil preparations are also anti-arrhythmic, particularly in patients who Chemicals have already suffered a myocardial infarction (MI) [9]. Acrylamide. The dose of ACR used was 45 mg/kg/ In addition to the treatment of hypertriglyceridae- /day [31]. Acrylamide (99% pure) was purchased from mia, a preparation of omega 3-acid ethyl esters is Sigma Chemical Company (St. Louis, Missouri, USA). licensed in the United Kingdom for the prevention It was dissolved in distilled water and administered of recurrent events after MI [32]. Recently, the com- every day by oral gavage. bined treatment of fish oil dietary supplement and Omega 3 plus. Omega 3 fatty acids (Sedico, omega 3 polyunsaturated fatty acids injections had Egypt) was given in a dosage of (200 mg/kg/day; been proved to promote post-traumatic brain injury DHA 100 mg/kg/day + EPA 100 mg/kg/day) which was (TBI) restorative processes in the brain, including administered every day by oral gavage [38]. generation of immature neurons, microvessels, and Green tea. Green tea. Royal Regime Tea, packed in oligodendrocytes, each of which was significantly Egypt by Royal Herbs, was dissolved in the drinking wa- correlated with the improved cognitive recovery [30]. ter at a concentration of 5 g/L. Tea was prepared freshly Green tea extract had been partially efficacious 3 times per week and stored at 48oC until use. The in preventing neurodegeneration in the brain of content of drinking vessels was renewed every day [23]. 565 Folia Morphol., 2019, Vol. 78, No. 3 Experimental design Dako autostainer instrument, EnVision Flex peroxi- The rats were randomly divided into four equal dase blocked. Immune reactions were evaluated with groups (6 rats each) as follows: various primary antibodies PDGFα (Genetex, USA), — Group I (control group): the rats received no med- BCL2 (Dako, Denmark), GFAP (DAKO, Denmark). Sec- ications; ondary antibodies used were Dako EnVision Flex/HRP. — Group II: the rats received ACR; Visualisation had been done with EnVision FLEX DAB — Group III: the rats received ACR concomitant with + chromogen. The prepared sections were examined omega 3; and photographed using a Canon digital camera — Group IV: the rats received ACR concomitant with (Canon, Japan) attached to the IBM computer system. green tea. All animals were sacrificed after 8 weeks (the end Histomorphometric study of the experiment) by decapitation using guillotines Image analysis was performed using the software to avoid brain injury [10]. Just before sacrification, the Leica Quin 500, Germany. It was used to measure body weight of all animals was measured. After eu- the areas per cent of positive PDGF, GFAP and BCL2 thanasia of animals, the cerebellum of all animals was immuno-staining reaction in a standard measuring immediately removed and underwent the following: frame using a magnification ×400 by light microscopy transferred to the monitor’s screen. These areas were Biochemical analysis masked by a green colour using the computer system. The cerebellum of all animals was immediately Area per cent values for each group were obtained washed in ice-cold glass slides, homogenised sepa- from 6 different fields from different slides. Values rately in 10 volumes (w/v) of 0.1 M phosphate buffer, were presented as a mean and standard deviation pH 7.4 using a polytron homogenizer for 1 min. The and statistically analysed. For optical density of silver homogenates were centrifuged at 4000 rpm for 20 min staining, the video signals from the video camera and refrigerated at 4°C. The supernatant was used for were transmitted to the image capture board which estimation of the quantitative activities of both malon- digitally converted light intensity to 256 possible grey dialdehyde (MDA) and the total glutathione (GSH).
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