Effect of Aqueous Extract of Khaya Grandifoliola C. DC. Stem Bark on Some Disaccharidases Activity in Iron-Deficient Weanling Rats
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Asian Journal of Research in Biochemistry 7(3): 1-7, 2020; Article no.AJRB.60874 ISSN: 2582-0516 Effect of Aqueous Extract of Khaya grandifoliola C. DC. Stem Bark on some Disaccharidases Activity in Iron-Deficient Weanling Rats Emmauel Gboyega Ajiboye1* and Temidayo Adenike Oladiji1 1Department of Biochemistry, University of Ilorin, Ilorin, Nigeria. Authors’ contributions This work was carried out in collaboration between both authors. Author EGA designed the study, performed the statistical analysis, wrote the protocol and wrote the first draft of the manuscript. Authors EGA and TAO managed the analyses of the study. Author EGA managed the literature searches. Both authors read and approved the final manuscript. Article Information DOI: 10.9734/AJRB/2020/v7i330138 Editor(s): (1) Dr. Mohamed Fawzy Ramadan Hassanien, Zagazig University, Egypt. Reviewers: (1) Eugenia Henriquez-D’Aquino, University of Chile, Chile. (2) B. Kumudhaveni, Madras Medical College, India. Complete Peer review History: http://www.sdiarticle4.com/review-history/60874 Received 27 June 2020 Original Research Article Accepted 03 September 2020 Published 08 September 2020 ABSTRACT This study was aimed at investigating the effect of the aqueous extracts of Khaya grandifoliola stem bark on some disaccharidases (lactase and sucrase) in diet-induced anemia in weanling rats. Weanling rats of 21 days old were maintained on iron-deficient diets for four weeks to induce anemia before treatment. A total of 35 weanling rats were used, grouped into five rats per group of iron-deficient diet/distilled water, iron-sufficient diet/distilled water, change of iron-deficient diet after four weeks to iron-sufficient diet, iron-deficient diet/ Standard drug, iron-deficient diet/25 mg/kg body weight of aqueous plant extract, iron-deficient diet/50 mg/kg body weight of aqueous plant extract, and iron-deficient diet/100 mg/kg body weight of aqueous plant extract. Phytochemical screening of the extract revealed the presence of alkaloids, saponins, cardiac glycosides, tannins, anthraquinones and flavonoids. The extract administered orally produced significant increase in the activity of intestinal mucosa sucrase and lactase (P < .05). The change of diet from iron deficient diet to iron sufficient diet increased disaccharidases activities in the intestinal mucosa. However, the aqueous extract of Khaya grandifoliola showed a higher disaccharidases activity when compared to the group of rats that were fed iron-sufficient diet. This study revealed that plant extract administered _____________________________________________________________________________________________________ *Corresponding author: E-mail: [email protected]; Ajiboye and Oladiji; AJRB, 7(3): 1-7, 2020; Article no.AJRB.60874 increased disaccharidases activity in the intestinal mucosa in diet-induced anemic group of weanling rats and thus lends credence to Khaya grandifoliola use in folklore medicine in the management of anemia. Keywords: Khaya grandifoliola; iron-deficient; lactase; sucrase. 1. INTRODUCTION 2. MATERIALS AND METHODS Iron deficiency anemia (IDA) is the most common 2.1 Experimental Animals type of anemia that results from an inadequate iron supply to aid the production of healthy red Forty-five weanling albino rats of both sexes blood cells [1]. Iron deficiency is usually a result (Rattus norvegicus) with a mean weight of 40.0 g of the depletion of the body's iron stores; hence, ±3.0 g were obtained from the Animal Holding a restricted supply of iron to various tissues Unit of the Department of Biochemistry, becomes apparent. This may result in a University of Ilorin, Nigeria. reduction in the synthesis of red blood cells and iron-dependent intra-cellular enzymes 2.2 Feed Components participating in many metabolic pathways [2]. Iron deficiency has been reported to reduce the Yellow maize (Zea mays) and locust bean activity of some disaccharidases [3,4]. (Parkia biglobosa A. Jacq) seeds were obtained Although there are various drugs used for the locally from Baboko Market, Ilorin, Nigeria, while treatment of anemia [5,6,7], they are not the soybean oil used was a product of Grand affordable to many poor people in the developing Cereals and Oil Mills Limited, Bukuru, Jos, countries such as Nigeria. Furthermore, the rural Nigeria. The vitamin mix was a product of BASF populations in various parts of the world do not Aktiengesellschaft, Germany Pantex, Netherland. have appreciable access to high-quality drugs for Component chemicals of the mineral mix used the treatment of anemia, hence, the reliance on were products of Sigma-Aldrich Chemical herbal products for the treatment of diseases and Limited, London, UK. anemia. Some of the plants with ethnobotanical and scientific claims being used for the treatment 2.3 Reagents of anemia include Waltheria indica, Sorghum bicolor, Khaya grandifoliola, and Mangifera indica The reagents used were of analytical grade and [8,9,10,11,12]. were prepared in all glass-distilled water. The reagents were stored in reagent bottles [17]. Khaya grandifoliola is a medicinal plant commonly found in Nigeria and many parts of the 2.4 Plant Identification and Preparation of plant are valuable to traditional medicine. Some Extract sicknesses like gastric pain, lumbago, stomach ache, worm infestation, cough, fever, and The stem barks of the plant were obtained from rheumatism are treated with concoctions made Ikaro in the Ose Local Government Area of Ondo from the bark of Khaya grandifoliola [13]. The State in the South West region of Nigeria. It was stem bark of this plant has also been previously authenticated in the Department of Plant Biology, reported to have anti-malarial, anti-ulcer, anti- University of Ilorin, Ilorin, Nigeria, where a hypoglycaemic, anti-hypoproteinaemia, and anti- voucher specimen was deposited in the hypocholesterolaemia activity [14,15,16]. Several herbarium with voucher number UILH/003/1066. research studies have lent scientific credence to the use of various parts of Khaya grandifoliola in The method described by Oladiji et al. [9] was the treatment of several diseases, however, used in the preparation of the plant extract. The there is still no adequate information on the effect stem barks of the plant were air-dried until a of the plant extract on lactase and sucrase constant weight was obtained. The dried pieces activity as an indicator for anti-anemic potentials were then pulverized using an electric blender, of Khaya grandifoliola. This study investigated and the powder obtained was stocked in a plastic the effect of aqueous extract of Khaya container. A known weight (15 g) of the powder grandifoliola stem bark on lactase and sucrase was poured into 100 ml of distilled water and activity in the intestinal mucosa of iron-deficient immediately boiled for 25 min. The resulting weanling rats. solution was left to cool and filtered with 2 Ajiboye and Oladiji; AJRB, 7(3): 1-7, 2020; Article no.AJRB.60874 Whatman filter paper. The filtrate was B – 35 weanling rats maintained on an iron- concentrated on a water bath (kept at 45°C) until deficient diet designated as IDF. a constant weight was obtained to give 2.32 g ± 0.03 of the residue (brownish-black slurry), which Animals in groups A and B were maintained on corresponded to percentage yield of 15.47%. their respective diets for four weeks. At the end The residues were pooled together and of the four weeks feeding period, five rats each reconstituted in distilled water to give the doses from ISF and IDF groups were sacrificed. The of 25, 50, and 100 mg/kg body weight. remaining weanling rats in groups B were further grouped into six with five rats in each group as The reconstituted aqueous extract was follows: administered orally using an oropharyngeal cannula to all the animals in different groups. B1- Iron deficient weanling rats fed with iron- deficient feed for two weeks (iron deficient 2.5 Diet Formulation feed all through) designated as IDF The composition of iron-deficient and iron- B2- Iron deficient weanling rats fed with iron sufficient diets per kg diet is described in Table 1. sufficient feed for two weeks (change of The components of the diets were thoroughly feed) designated as COF mixed and made into pellets to ensure homogeneity and proper handling by the B3- Iron deficient weanling rats orally animals. It was produced weekly and packed into administered daily for two weeks with iron air-tight polythene bags to prevent rancidity, supplement syrup (a standard iron auto-oxidation of the oil, and microbial supplement) designated as SD contamination. The proximate analysis of the compounded feeds was also carried out. B4- Iron deficient weanling rats orally administered daily for two weeks with 25 2.6 Animal Grouping and Extract mg/kg body weight of aqueous extract of Administration Khaya grandifoliola stem bark designated as KG 25 mg. The animals were kept in well-ventilated house conditions (temperature: 22±3°C; photoperiod: B5- Iron deficient weanling rats orally 12 h/12 h light/dark cycle; relative humidity: 45- administered daily for two weeks with 50 50 %). They were allowed free access to normal mg/kg body weight of aqueous extract of rat chow and distilled. The acclimatization was Khaya grandifoliola stem bark designated as done for seven days, and they were then fasted KG 50 mg. for 24 hours (without food but water) before the B6- Iron deficient rats orally administered commencement of the experiment. The animal daily for two weeks with 100 mg/kg body grouping consisted of an initial two groups: weight of aqueous extract of Khaya A – 10 weanling rats maintained on iron- grandifoliola stem bark designated as KG sufficient diet designated as ISF. 100 mg. Table 1. Feed components of iron sufficient and iron-deficient diets Feed components Iron-sufficient (g/kg) Iron-deficient (g/kg) Locust bean seed 500 500 Maize flour 315 315 Soybean oil 40 40 Sucrose 100 100 Methionine 5 5 Mineral mixb 30 30 Vitamin mixa 10 10 FeSO4·7H2O 157.36mg/Kg 38.216 mg/Kg Soybean oil: polyunsaturated fatty acids (58%), monounsaturated fatty acids (29%), saturated fatty acids (13%).