Brazilian Journal of Biological Sciences, 2017, v. 4, No. 7, p. 67-80. ISSN 2358-2731 https://dx.doi.org/10.21472/bjbs.040708 Antihyperlipidemic and hypolipidemic properties of Tacca leontopetaloides (L.) Kuntze (Dioscoreales: Dioscoreaceae) tuber’s aqueous extracts in the rats Doubla Sali Aïssatou¹, Josiane Thérèse Ngatchic Metsagang²,*, Celestin Dongmo Sokeng¹ and Nicolas Yanou Njintang¹ ¹Department of Biological Sciences, Faculty of Sciences, Ngaoundere University, P.O. Box 454, Ngaoundere, Cameroon. ²Department of Food Science and Nutrition, National School of Agro-Industrial Science, P.O. Box 455 Ngaoundere, Cameroon. *Email: [email protected]. Abstract. Tubers of Tacca leontopetaloides (L.) Kuntze (Dioscoreales: Dioscoreaceae) play an important role in food Received substitution during the periods of food shortage in occidental and February 23, 2017 central Africa. It is also used in traditional medicine for the treatment of the diarrhea, dysentery, stomach evil, viral hepatitis and Accepted the infection of guinea worn. This study attempted to evaluate the May 30, 2017 antihyperlipidemic and hypolipidemic properties of aqueous extracts of the tubers of T. leontopetaloides in rats. For this experimentation, Released two tests were done: antihyperlipidemic and hypolipidemic tests. Hyperlipidemia was induced in rats with High Fat Diet containing June 30, 2017 300 g of egg yolk, 2 g of cholesterol, 250 g of coconut oil and 50 g Open Acess of soya oil. The group of rats on which the antihyperlipidemic test Full Text Article was done was fed with High Fat Diet and supplemented with T. leontopetaloides aqueous extract at 27.34 mg/kg, and 54.68 mg/kg; after 21 days of experimentation, the different groups of rats were sacrificed. The lipid profile and some biochemical parameters were evaluated. Organs like kidneys, liver and stomach were taken for histopathological evaluation. For the hypolipidemic test, after three weeks of induction of hyperlipidemia, the diet was changed to normal diet and aqueous extract of tubers was given to rats during 21 days at doses of 3.41 mg/kg and 13.67 mg/kg. The animals were sacrificed after 21 days of experimentation. The lipid profile, some biochemical parameters and histopathology of organs were evaluated. The antihyperlipidemic and hypolipidemic tests of ORCID aqueous extract of the tubers of T. leontopetaloides induced 0000-0002-0775-2198 reduction in total cholesterol, triglyceride, LDL-cholesterol and an Doubla Sali Aïssatou increase in HDL-cholesterol significantly (p < 0.05); decreasing 0000-0002-5385-5518 activities of ALAT and ASAT enzymes, levels of creatinin remained Josiane Thérèse no changed. Histopathological study revealed that extracts did not Ngatchic Metsagang repair the destructions of liver cells and glomerules of kidneys 0000-0003-3713-059X caused by High Fat Diet. Aqueous extract of tubers of Celestin Dongmo Sokeng T. leontopetaloides exhibits antihyperlipidemic and hypolipidemic 0000-0002-8949-7150 activities; hence it could be important in the management of Nicolas Yanou Njintang cardiovascular diseases. ISSN 2358-2731/BJBS-2017-0005/4/7/8/67 Braz. J. Biol. Sci. http://revista.rebibio.net 68 Aïssatou et al. Key words: Tacca leontopetaloides, antihyperlipidemia, hypolipidemia, saponins. Introduction Materials and methods Hypercholesterolemia and Plant material and preparation of hypertriglyceridemia are two major risk the extract factors of cardiovascular diseases. They The tubers of T. leontopetaloides accelerate the development of coronary were collected in the locality of Karna- artery diseases and the progression of Manga in the Adamaoua Region of atherosclerosis (Ballantyne, 2007; Hamlat Cameroon and were transported at the et al., 2008). High levels of low-density laboratory of Biophysics, Food lipoprotein (LDL) accumulate in the Biochemistry and Nutrition of Ngaoundere extracellular and sub-endothelial space of University. The tubers were washed and arteries are highly atherogenic and toxic to rinsed with tap water.Thereafter, the tubers vascular cells there by leading to were peeled and sliced (sections of 7 mm atherosclerosis, hypertension, diabetes, Ø) using a chips slicer (Potato-chipper), functional depression in some organs 300 g of the sliced tubers were crushed with (Suresh, 2001; Saghir et al., 2012). Tacca 3 L of distilled water using Moulinex leontopetaloides is a wild plant belonging (model Microton MB 550, Kinematica AG, to the family of Taccaceae. It found in the Switzerland). Lastly, the crushed tubers was tropical mediums, particularly in Occidental centrifuged at 4,000 tours/min during and Central Africa (Kay, 1987). The tubers 20 min, at 25 °C. The supernatant obtained of T. leontopetaloides have an important was centrifuged again under the same energetic value and play a crucial role in conditions as previously described and the food substitution during the periods of food second supernatant constitutes aqueous shortage in Africa (Kay, 1987; Garine, extract. 2002). It is used in traditional medicine for the treatment of the diarrhea, dysentery and Chemical composition evils of stomach, viral hepatitis and the determination infection of Guinea worm (Kay, 1987; Total phenol content of extract was Garine, 2002). Studies of Ndouyang et al. determined as gallic acid equivalents (2009) showed that the tubers of according to the method described by T. leontopetaloides were rich in saponins Makkar et al. (1993) with some (10.78%). Several studies have shown modifications. After extraction with 70% hypolipidemia properties of saponins (v/v) methanol, in the same extract, total (Francis et al., 2002; Reynolds et al., 2006; tannin content was determined by the Afrose et al., 2010). In spite of the fact that precipitation method using T. leontopetaloides tubers content saponins, polyvinylpyrrolidone (PVPP) as described no work has been interest on their effect on by Makkar et al. (1993) with some lipidemia. modifications. PVPP in extract binds to Thus this study aim to evaluate the tannins and make them inert. Briefly in antihyperlipidemic and hypolipidemic 100 mg PVPP, 1.0 mL of distilled water properties of aqueous extracts of tubers of and 1.0 mL of sample extract were added. T. leontopetaloides. Braz. J. Biol. Sci., 2017, v. 4, No. 7, p. 67-80. Antihyperlipidemic and hypolipidemic properties of Tacca leontopetaloides 69 The blend was vortexed and kept at 4 °C for of aqueous extract having been treated. The 15 min, vortexed once more and alkaloid content was calculated by the centrifuged at 3,000 trs/min for 10 min at following formula: 25 °C. The supernatant composed of only simple phenolics other than tannins were collected. The phenolics content of the supernatant were determined as mentioned above and the content of non-tannin phenolics expressed. The tannin content of Animal experimentation and the sample was calculated as difference induction of hyperlipidemia between total phenolics and non-tannin Male adult Wistar albinos rats (200- phenolics. The flavonoids were determined 250 g) were obtained from the animal house by the method describe by Adom et al. of National School of Agro-Industrial (2003). A volume of 0.1 mL of methalonic Sciences of the University of Ngaoundere. extract was introduced in solution prepared The animals were used for both tests, with 5% sodium nitrate and 10% antihyperlipidemic and hypolipidemic. hexahydrated aluminium chlorite Hyperlipidemia was induced by High Fat ACL36H2O. The DO was read at 510 nm. Diet (HFD) containing 300 g of egg yolk, 2 Saponins were evaluated according g of cholesterol, 250 g of coconut oil and to the method of Hiai et al. (1976) with 50 g of soya oil, as described by Hamlat et some modifications. 5 mL of aqueous al. (2008) with some modifications. Table 1 extract of tubers were added to 5 mL of shows the different formulations. methanol (80%). The mixture was homogenized and centrifuged at 3000 Antihyperlipidemic test trs/min, for 10 min at 25 °C. The Thirty Wistar albinos’ male rats supernatant was recovered and evaporated weighing 200-250 g rats were randomly at 50 °C in rotary evaporator until divided into 5 groups of 6 rats each. The evaporation of methanol. To 0.1 mL of rats were housed in cages in a room where supernatant obtained, 0.25 mL of methanol the temperature was 37 °C and 12 h light (80%) were added, 0.25 mL of vanillin and dark cycles were maintained and water (8%) and 2.5 mL of sulphuric acid (72%) was given ad libitum. The first group fed were added slowly on the internal wall of with HFD (negative control, GI), the second the tube. The content of the tube was group fed with HFD and atorvastatin at 10 homogenized carried at 60 °C in the bain- mg/kg (positive control, GII), the third marie. After 10 min of incubation, the tubes group fed with HFD and aqueous extract of were cooled in tap water during 3-4 min, tuber of T. leontopetaloides at the dose of and read at 544 nm against white tube. 27.34 mg/kg (GIII), the fourth group was The alkaloid was determined fed with HFD and aqueous extract of tuber according to Harborne (1991) methods with of T. leontopetaloides at the dose of 54.68 some modifications. A volume v of 20 mL mg/kg (GIV) and the five group was fed of aqueous extract was centrifuged and with standard diet (normal control, GV). filtered, to the aqueous filtrate; ammonia The animals were weighed each week, the (40%) solution was added until obtain pH quantity of feed intake was obtained by 8-9. Then this volume was extracted with subtracting the remaining feed from the 20 mL of chloroform and the two phases quantity administered the previous days. were separated using a decant ampoule. Feces were collected daily for the Thus the chloroform phase obtained was evaluation of the excreted lipids. After three filtered in a becher and the weight was weeks, the animals were sacrificed by taken (M0). The bechers containing the anesthesia using diethyl ether, and the samples were left in the drying oven 103 °C blood samples were collected from the to enable chloroform evaporation.