Hepatoprotective Activity of Hyphaene Thebaica Extract Against Mercuric
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Progress in Nutrition 2017; Vol. 19, Supplement 1: 142-149 DOI: 10.23751/pn.v19i1-S.5083 © Mattioli 1885 Original Article Hepatoprotective activity of Hyphaene thebaica extract against mercuric chloride-induced hepatotoxicity in adult male albino rats Safia Mohamed Shehata1, Eman Ali Abd El-Ghffar2 1 Clinical Pathology Department, Ain Shams University Hospitals, Cairo, Egypt; 2Zoology Department, Faculty of Science, Ain Shams University, Khalifa El-Maamon st., Abbasiya sq., 11566, Cairo, Egypt E-mail address: [email protected] Summary. Doum, Hyphaene thebaica (H. thebaica), known as a famine food, is an African palm tree common in Upper Egypt. The present study was performed to evaluate the protective effect of H. thebaica (the fruits of doum) extract on liver/kidney functions in mercuric chloride (HgCl2) treated rats. Rats were randomly clas- sified into healthy control, and 5 treated groups which were H. thebaica extract alone with either 500 or 1000 mg/kg b.w, HgCl2 alone, H. thebaica extract (low dose) plus HgCl2 and H. thebaica extract (high dose) plus HgCl2 groups. Results clearly revealed that consumption of H. thebaica alone (especially high dose) showed improvement of healthy status. The pre-treatments of H. thebaica extract with high dose which had the best modulatory results as increasing hepatic non-enzymic/enzymic antioxidant system as well as decreasing pro- inflammatory cytokines compared with healthy control group with decreasing lipid peroxidation. Our results indicate that consumption of H. thebaica extract has a beneficial role against hepatotoxicity induced by HgCl2 in male albino rats. Key words: Doum, HgCl2, hepatotoxicity, rats Introduction carp and the outer coat of the endocarp of the fruits of doum are inedible, while the mesocarp and kernel Heavy metal such as mercury (Hg) are considered flesh of the fruits are edible. Research on the fruit pulp as the most dangerous substances causing many health of H. thebaica showed that it contains nutritional trace hazards to human and animals through progressive ir- minerals, proteins and fatty acids, in particular the nu- reversible accumulation in their bodies. Hg is capable tritionally essential linoleic acid (4). The identification of damaging the organism in several ways due to its of compounds, by thin-layer chromatography, showed high affinity to various tissues such as liver, kidney, that the fruit pulp of H. thebaica contains significant brain, reproductive organs, lung, immune system and amounts of saponins, coumarins, hydroxycinnamates, its tendency to accumulates (1,2). Recent evidences essential oils, flavonoids, phenolics acids and oxylip- also show that Hg causes severe oxidative changes (3), ids. In addition, 17 compounds were identified and thus Hg proved to be a potential oxidant is the catego- quantified from the fruit of H. thebaica, including 2 ry of environmental factories. Therefore, there is a need cinnamic acid derivatives, 5 flavonoids, 6 fatty acids, to provide protection against Hg induced toxicity. 2 sphingolipids, a lignan, and a stilbene. The plant’s Hyphaene (H.) thebaica is one of the plants used sugar composition was characterized and quantified by in ethno-medicine and belongs to the family Palm and 1H-nuclear magnetic resonance, with sucrose detected subfamily Borassoideae. It grows commonly in both as its major component at a level of 219 mg/g (5, 6). Sahel and Sahara regions in Upper Africa. The peri- H. thebaica also contains crude protein and lipids (7, 8). Hepatoprotective activity of Hyphaene thebaica extract against mercuric chloride-induced hepatotoxicity in adult male albino rats 143 Also, the aqueous extract of H. thebaica fruits showed polypropylene cages with wood dust and given stand- an antioxidant activity; this is due to the substantial ard rodent food pellets (Agricultural-Industrial Inte- amount of their water-soluble phenolic contents of fla- gration Company, Giza, Egypt) and tap water ad libi- vonoids within it, which represent conjugates of o-gly- tum. The animals were procured, maintained and used cosides and include quercetin, chrysoeriol, luteolin, and in accordance with WHO guideline for animal care isorhamnetin H (9-11). Also, H. thebaica is thought to and the study design was approved by the Ain Shams possess anti-inflammatory property due to its ability to University Research Ethics Committee. inhibit cyclooxygenase (12). Locally, various extracts and decoction of fruit pulp of H. thebaica are used in Experimental design. After one week of acclima- the treatment of bilharzia, haematuria, hypertension tization, all animals were randomly divided into six and as a haematinic agent in human and in animal groups of five rats each: models (4,13). Also, it has a cooling and soothing ef- Group I served as healthy control and orally re- fect on the digestive system and antioxidant proper- ceived 1 ml double distilled water once daily for 28 ties which help flush toxins from the body (11,14). In days, then on the 28 day and after 2 hours from last addition, H. thebaica may be advised as a good choice dose of distilled water administration, rats were sub- that can delay diabetic complications in rats (6). This cutaneously injected with 0.5 ml normal physiological study was undertaken to study the protective effect of saline at once. fruit pulp of H. thebaica on hepatotoxicity induced by Group II served as control treated and orally re- HgCl2 in male rats and to find its probable mechanism ceived 1 ml of H. thebaica extract at dose of 500 mg/kg of action including its antioxidant potential. b.w once daily for 28 days. Group III served as control treated and orally re- ceived 1 ml of H. thebaica extract at dose of 1000 mg/ Materials and methods kg b.w once daily for 28 days. Group IV (Hg-treated) rats were given a single Plant material. The fruit of H. thebaica (doum) subcutaneously injection of Hg in the form of HgCl2 at was obtained from local markets (Cairo, Egypt) and dose of 5 mg/kg b.w dissolved in normal physiological was authenticated by Dr. U.K. Abdel-Hameed, a plant saline (15) on the 28 day of the experiment. taxonomist in the botany department, faculty of sci- Group V (H. thebaica + Hg-treated group) re- ence, Ain Shams University, Cairo, Egypt. A voucher ceived orally 1ml of H. thebaica extract (dissolved in specimen was deposited in the herbarium of the botany double distilled water) at dose of 500 mg/kg b.w/day department, faculty of science, Ain Shams University. (13) for 28 days, then, after 2 hours from last dose of extract administration, rats were treated with HgCl2 Preparation of suspension. The fruits of H. thebaica (dose same as that in the Hg-treated group). was cleaned and separated into pulp. The pulp was then Group VI (H. thebaica + Hg-treated group) re- dried and ground into fine powder using mortar and ceived orally 1ml of H. thebaica extract (dissolved in pestle. Subsequently, 5g of the powder was extracted double distilled water) at dose of 1000 mg/kg b.w/ by 70% ethanol on cold until exhaustion. In order to day for 28 days, then, after 2 hours from last dose of obtain the dried extracts, the solvent was distilled in extract administration, rats were treated with HgCl2 rotary evaporator at 55°C till dryness. Aqueous sus- (dose same as that in the Hg-treated group). pension was performed by suspending in double dis- At the end of the experimental period (day 29), tilled water for final administration. the rats were sacrificed; blood, liver and kidney sam- ples were collected for different analyses. Animals. Adult male albino rats (Rattus norvegi- cus), weighing about 130-140 g, were obtained from Measurements. Body weight (gain or loss) was cal- the Animal House Colony of the National Research culated by the following equation: body weight (gain Center (Dokki, Giza-Egypt). Animals were housed in or loss) = body weight at the end of the experiment ̶ 144 S. M. Shehata, E. A. Abd El-Ghffar body weight at the beginning of the experiment. The (w/v: 0.5 g tissue with 5 ml PBS, PH 7.4). Homogen- relative organs weight (liver and kidney) was calculat- ates were centrifuged at 10.000 rpm for 15 min at 4°C. ed according to the following equation: relative weight The resultant supernatant was used for determina- (g/100 g b.w) = [(organ weight at the end of the ex- tion of the activities of liver catalase (CAT) (21), glu- periment × 100) / (animal body weight at the end of tathione peroxidase (GSH-Px) (22) and glutathione- the experiment)]. S-transferase (GST) (23), as well as the levels of reduced glutathione (GSH) (24) and malondialdehyde Serum Preparation. Blood samples were taken by (MDA) (25) using Bio-diagnostic kits (Bio-diagnostic cutting the neck at the jugulars by a sharp razor blade Company, Giza, Egypt). after the rats were subjected to light diethyl ether an- aesthesia without anticoagulant to separate serum by Statistics. All numeric variables were presented as centrifugation in a cooling centrifuge (IEC centra- mean with ± standard error (SE). Statistical compari- 4R; International Equipment Co., Needham Heights, sons were performed using one-way ANOVA using MA, USA) for 30 min at 3000 rpm and 4°C. The serum Graph Pad Prism version 4.03 for Windows (Graph was separated and divided into samples and preserved Pad Software Inc., San Diego, CA, USA). The differ- at -80°C for the subsequent estimation of biochemical ence between means was assessed by Tukey’s multiple parameters. The appropriate kits (Bio-diagnostic kits) comparison test (26), in which P values of <0.05, <0.01 were used for determination of the activities of se- and <0.001 were considered statistically significant, rum alanine and aspartate aminotransferases (ALAT, highly significant and very highly significant, respec- ASAT) (16), total protein level (17), albumin level tively.