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Evaluation of the Anti-Gastric Ulcer Effect of Methanolic Extract of Pennisetum Purpureum (Schumach) in Male Wistar Rats

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Evaluation of the Anti-Gastric Ulcer Effect of Methanolic Extract of Pennisetum Purpureum (Schumach) in Male Wistar Rats Int.J.Curr.Microbiol.App.Sci (2015) 4(10): 466-474 ISSN: 2319-7706 Volume 4 Number 10 (2015) pp. 466-474 http://www.ijcmas.com Original Research Article Evaluation of the Anti-Gastric Ulcer Effect of Methanolic Extract of Pennisetum purpureum (Schumach) in Male Wistar Rats Onwuchekwa Chinedu* and Aliyu Buhari Department of Physiology, Faculty of Basic Medical Sciences, College of Health Sciences, Usmanu Danfodiyo University, Sokoto Nigeria *Corresponding author A B S T R A C T The fresh leaf samples of Pennisetum purpureum has been shown to be a potential source of dietary protein and antioxidants. The proximate composition and selected K e y w o r d s micronutrients of young leaf sheaths with their edible inner whorled leafy bundles collected from Napier grass in Nigeria (achara Ibeku variety, with purple sheaths) Pennisetum has been determined from previous studies. There is dearth of information on purpureum, possible anti-ulcerogenic effect of methanolic extract of edible young leaves of Gastric ulcer, Napier grass (Pennisetum purpureum). In this study, the anti-gastric acid effect of Mucus, fresh leaf samples of achara Ibeku was determined by inducing ulcer with ethanol. Malonialdehyde The gastric mucus secretion, mucus cells count and malonialdehyde concentration were also determined. The study showed that the methanolic extract of Napier grass (Pennisetum purpureum) ameliorated gastric ulcer effect. Introduction Pennisetum purpureum (Schumach) that are respectively known in local Igbo commonly known as elephant grass or dialects as Achara Ibeku and Achara Ngwa Napier grass, is a tall grassy perennial plant cultivars. The naturally sheathed tender in Poaceae family (Obi et al., 2008; Cowell, young leaves of Napier grass which is 2009; Feedipedia, 2015). Though a native of cherished as a soup vegetable especially sub- tropical Africa, it now exists in most amongst Ngwa and Umuahia-Ibeku clans of tropical and sub-tropical countries of the Igbo people of Abia State, Nigeria world as a weed or planted crop that is (Okaraonye and Ikewuchi, 2009) and some largely produced and used for livestock feed of their surrounding neighbors is also known (Obi et al., 2008). The apical ends of culms as Achara in the local Igbo dialects. The or stems of Napier grass plants usually have ethnic soup that is prepared with this leafy whorled tender young leaves before vegetable is called Ofe Achara flowering (Cowell, 2009). The common two (Okaraonye and Ikewuchi, 2009). The Igbo varieties of the plant found in Nigeria are people consume this soup with those with purple stem and light green stem; complimentary ethnic viscous starchy pastes 466 Int.J.Curr.Microbiol.App.Sci (2015) 4(10): 466-474 such as yam fufu and eba (Ukpabi and 6o 14 N, and between longitude 7o 10 Ndimele, 1990; Ukpabi and Oti, 2010). and 8o E). These botanic shoot samples were Presently, this perishable leafy vegetable authenticated as materials from Napier grass from Napier grass is being marketed in (Pennisetium purpureum) by Botany Unit, Nigeria. Furthermore, the nutritional data on Department of Plant Science and this leafy vegetable did not indicate varietal Biotechnology, Michael Okpara University effect on its nutritional composition of Agriculture, Umudike, Abia State, (Okaraonye and Ikewuchi, 2009) and Nigeria. The exterior parts of the sheaths another study indicated that the were hand peeled or rolled off to get the soft experimental fresh leaf samples of Achara edible part or matrices also known as Ibeku variety are potential sources of dietary Achara . 1000g of these were placed in protein and antioxidants. The dearth of Soxhlet extractor and extracted using information on possible gastrointestinal methanol. The extract was concentrated with effect viz-a-viz gastric ulcer of this local the aid of rotary evaporator (Heidolph, food and feed stuff led to this study. model number 4011, USA) to give a yield of 20g of dry extract. Materials and Methods Acute toxicity study of the extract Chemicals Adult albino mice (25- 30g) were divided Histamine acid phosphate (Sigma- Aldrich, into five groups each containing 10 mice. St Louis MO), Ketamine Hydrochloride The mice were fasted for 6 h with only (Rotexmedica, Trittau, Germany), access to water ad libitum before the Carbamylcholine chloride (Carbacol: experimental study. Groups I, II, III and IV Sigma- Aldrich MO), Pentagastrin (Sigma animals were administered various doses of Aldrich), Trichloroacetic acid, Magnesium methanolic extract of Pennisetum chloride (Sigma- St Louis, MO), Sodium purpureum (500, 800, 1000 and 1500 chloride (Sigma- St Louis, MO), mg/kg) orally and Group V received Tween Hydrochloric acid (Sigma- St Louis, MO), 80 only. The animals were observed for 72 h Thiobarbituric acid (Sigma- St Louis, MO), for mortality (Ravichandran et al., 2007). Alcian blue, Sodium acetate(Sigma- St There was no death during and after the test. Louis, MO), Diethyl ether (Sigma- St Louis, MO), Sodium hydroxide (Sigma- St Louis, Ethanol induced ulcer model MO). Chemicals and reagents were of analytical grade. The ulcer was induced by administering ethanol. All the animals were divided into Source of materials and extraction of four groups each consisting of five rats. plant material Group I received distilled water orally and served as control. Groups II and III rats Samples of the experimental young shoots received Pennisetum purpureum extract of Achara Ibeku variety (with purple colour (EPP) dissolved in Tween 80 at doses of 200 leaf sheaths) leafy materials covered with and 400 mg/kg respectively. Rats in Group tough leaf sheaths were purchased at Orie IV were administered with omeprazole (20 Ugba market, Umuahia North local mg/kg) as a standard reference drug. Rats in government area in Abia state of Nigeria Group IV were administered with (approximately within latitude 4o 40 and omeprazole (20 mg/kg) as a standard 467 Int.J.Curr.Microbiol.App.Sci (2015) 4(10): 466-474 reference drug. The gastric ulcers were weight of the stomach, to give the weight of induced in the rats by administering mucus secreted. 1ml/200g of absolute ethanol (90%) after 45 minutes of EPP and omeprazole treatment. Thus, gastric mucus secretion (mg/g tissue) They were kept in modified cages to prevent Weight of dye (mg) coprophagy during and after the experiment. = The animals were anaesthetized 1 h later Weight of stomach (g) with ketamine (0.2 ml/100g body weight). The stomach was removed and incised along Effect of EPP on gastric mucus cell count the greater curvature and ulceration scored using the method of Desai et al. (1999). The rat were divided into four sub-groups of five rats each and treated for 14 days orally Effect of EPP on gastric mucus secretion with EPP. Group 1 was the control and was treated with distilled water. Groups II and III The rats were divided into four sub-groups were given EPP at doses 200 mg/kg and 400 of five rats and treated for 14 days orally mg/kg respectively, while Group IV was with extract of EPP. Group 1 was the control administered omeprazole (20 mg/kg). The treated with distilled water and Group IV rats were sacrificed by cervical dislocation was administered omeprazole (20 mg/kg). and the stomachs removed. The glandular Groups II and III were given EPP at doses portion of the each stomach was opened 200 mg/kg and 400 mg/kg respectively. The along the lesser curvature. Gastric mucus rats were sacrificed by cervical dislocation cell count was done by counting the number and their stomachs removed and weighed. of gastric mucus cells that stained with The glandular portion of each stomach was haematoxylin and eosin. The stained slide of opened along the lesser curvature. The the stomach mucosa viewed were indicated everted stomachs were soaked for two hours as blue patches. These were counted using in 0.1% Alcian blue dissolved in 0.16M calibrated microscope in five randomly sucrose buffered with 0.05M sodium selected area of the gastric mucosal tissue. acetate, adjusted to pH 5.8 with hydrochloric Five cubic boxes each with an area of 1 mm2 acid. were assessed. This method is an improvement over the earlier described Uncomplexed dye was removed by two approach for counting by Li et al. (2002). successive washes at 15 and 45 minutes in 0.25M sucrose. Dye complexed with mucus Effect of EPP on Malondialdehyde was diluted by immersion in 10ml aliquots Concentration (MDA) of 0.5M Magnesium Chloride for 2 hours. The resulting blue solutions were shaken The rat were divided into four sub-groups of briefly with equal volume of diethyl ether five rats each and treated for 14 days orally and absorbance of aqueous phase was with EPP. Group 1 was the control and was measured at 605 nm using treated with distilled water. Groups 2 and 3 spectrophotometer (Corney et al., 1974). were given EPP at doses 200 mg/kg and 400 The absorbance of each solution was then mg/kg respectively, while Group 4 was used to calculate the various concentrations administered omeprazole (20 mg/kg). The of dye and the weight of dye (expressed in rats were sacrificed by cervical dislocation mg) deduced, using a standard curve. The and the stomachs removed. The stomach weight of dye was then expressed over the was opened along the lesser curvature, 468 Int.J.Curr.Microbiol.App.Sci (2015) 4(10): 466-474 washed and homogenized to be used for mg/kg (3.38 ± 0.16) and omeprazole 20 lipid peroxidation. Briefly, lipid mg/kg (2.25±0.19) compared to the control peroxidation was assessed by measuring (6.25 ± 0.43). It shows a graded and dose- thiobarbituric acid reactive substances dependent decrease in the mean ulcer scores (TBARS) produced according to the method with increases in dose of EPP.
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