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Conocarpus Lancifolius Malik Saadullah, Bashir Ahmad Chaudary, Muhammad Uzair and Khurram Afzal

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Conocarpus Lancifolius Malik Saadullah, Bashir Ahmad Chaudary, Muhammad Uzair and Khurram Afzal A Journal of the Bangladesh Pharmacological Society (BDPS); www.bdps.info Bangladesh J Pharmacol 2014; 9: 244-249 Journal homepage: www.banglajol.info Abstracted/indexed in Academic Search Complete, Agroforestry Abstracts, Asia Journals Online, Bangladesh Journals Online, Biological Abstracts, BIOSIS Previews, CAB Abstracts, Current Abstracts, Directory of Open Access Journals, EMBASE/Excerpta Medica, Google Scholar, HINARI (WHO), International Pharmaceutical Abstracts, Open J-gate, Science Citation Index Expanded, SCOPUS and Social Sciences Citation Index ISSN: 1991-0088; DOI: 10.3329/bjp.v9i2.18556 Antidiabetic potential of Conocarpus lancifolius Malik Saadullah, Bashir Ahmad Chaudary, Muhammad Uzair and Khurram Afzal Faculty of Pharmacy, Bahauddin Zakariya University, Multan, Pakistan. Article Info Abstract Received: 9 April 2014 The antidiabetic activity of Conocarpus lancifolius extract was investigated in Accepted: 6 May 2014 vitro , as alpha glucosidase inhibition and in vivo as alloxan induced diabetic Available Online: 7 June 2014 rabbits with other biochemical parameters (LDL, HDL, SGPT, SGOT, cretinine, Keywords: urea and triglyceride). Alpha-glucosidase inhibition activity was performed by Alloxan using acorbose as standred. Methanolic extract show alpha-glucosidase Alpha glycosidase inhibition activity. The dose of 200 mg/kg body weight significantly (p<0.05) Antidiabetic decreases the blood glucose level, plasma total cholesterol, triglycerides and Conocarpus lancifolius LDL in treated rabbits as compared to diabetic rabbits. This dose significantly Number of Tables: 6 increased the level of HDL in treated group. The activity of SGOT and SGPT Number of Refs: 35 also significantly (p<0.05) decreased in treated diabetic rabbits. Phytochemical Correspondence: MS studies show the presence of glycosides, tannins, saponins and terpenoids. e-mail: [email protected] The antidiabetic potential is may be due to its saponin contents. Introduction Materials and Methods World Health Organization estimated that for health Collection and extraction of plant material: The plant care needs, four-fifth of the total populations till confi- material was collected from surroundings of Lahore dence on the plant medicine ( Farnsworth et al., 1985 ). (Pakistan). The plant was identified as C. lancifolius Among the plants, Carica papaya (Sadeque and Begum, Engl . by Professor Dr. Altaf Ahmad Dasti, Taxanomist 2010 ), Carissa spinarum (Hegde and Joshi, 2010 ), Cestrum Institute of pure and applied Biology, Bahauddin nocturnum (Qadir et al., 2014 ), Chenopodium mura- Zakariya University, Multan. le (Saleem et al., 2014 ), Cocculus hirsutus (Thakare et al., For the purpose of effective extraction, whole plant of 2009 ), Convolvulus arvensis (Ali et al., 2013 ), Dodonaea vis the C. lancifolius was dried under the shade for 17 days. -cosa (Khan et al., 2013 ), Khamira Gaozaban Ambri Jad- The plant material was coarse grounded and then war Ood Saleeb Wala ( Akhtar et al., 2013 ), OfIpomoea grounded to fine powder. This powder was subjected to staphylina (Bag and Mumtaz, 2013 ), Suaeda fruticosa extraction through the simple maceration method at (Rehman et al., 2013 ), Trianthema decandra (Balamuru- room temperature occasionally shaking for 24 hours. gan and Muthusamy, 2008 ) and Trichodesma sedgwickia- The weighted amount (1 kg) of plant material was kept num (Saboo et al., 2013 ) showed hepatoprotective effect. in large size well caped containers and added 1200 mL Conocarpus lancifolius Engl ., an ornamental tree, is native of Methanol as solvent, to get the maximum extraction to coastal and riverine areas of East Africa ( Baroon et al., results; the flask having mixture was placed in 2012 ). The leaves are glossy in appearance with relatively ultrasonic bath for about half hour daily. Filtration was fewer trichomes on both surfaces ( Redha et al., 2011 ). done after the addition of solvent. Same process was The antidiabetic capacities of C. lancifolius consumed carried out 3 times to get the maximum extracted locally in Pakistan have not been presented. However, volume. The methanol (18.3 g) extract was collected in up to now a little phytochemical or pharmacological separate sample bottles and designated with code as work was done. CLM. This work is licensed under a Creative Commons Attribution 3.0 License. You are free to copy, distribute and perform the work. You must attribute the work in the manner specified by the author or licensor. Bangladesh J Pharmacol 2014; 9: 244-249 245 Phytochemical screening: The presence of alkaloids, another 20 min. Then the reaction was stopped by glycosides, tannins, flavonoids, anthraquinones and adding 160 µL of 0.2 M NaCO 3 into each well, and saponins were screened according to the method of absorbance readings (A) were recorded at 405 nm by (Trease and Evan, 1998 ). micro-plate reader and compared to a control which had 60 µL of buffer solution in place of the extract. For Alkaloids: about 5 g of powdered drug was boiled in blank incubation (to allow for absorbance produced by dilute hydrochloric acid. It was filtered and made the extract), enzyme solution was replaced by buffer alkaline by the addition of dilute ammonia solution. solution and absorbance recorded. The concentrations This alkalinized solution was extracted with 5 mL of of test compounds which inhibited the hydrolysis of chloroform. The chloroform layer was then extracted substrates (butyrylthiocholine) by 50% (IC 50 ) were with 10 mL dilute acetic acid. To the acetic acid extract, determined by monitoring the effect of increasing few drops of Dragendorff’s reagent was added (Orange concentrations of these compounds in the assays on the precipitate or turbidity indicate the presence of inhibition values ( Rehman et al., 2013 ). The IC 50 values alkaloids). were then calculated using the EZ-Fit Enzyme Kinetics Braemer’s test for tannins: 5 g of drug was extracted program (Perrella Scientific Inc., Amherst, USA). with methanol. To methanolic extract, 10%alcoholic ferric chloride solution was added. Dark blue or In vivo antidiabetic study greenish grey coloration of the solution indicates the Animal selection and treatment: 30 male rabbits each of presence of tannins in the drug. weighing 1-1.5 kg were used and all rabbits were Saponins: 5 g of powdered drug was vigorously shaken checked for evidence of any infections. All the rabbits with water and noted for production of froth (persistent were housed in the animal house of the Department of froth for 20 min indicates the presence of saponins). Pharmacy B.Z.U Multan. They were housed in the steel cages under standard laboratory condition (light period Borntrager’s test for free anthraquinones: 5 g of 8:00 am to 8:00 pm 21± 2 °C, relative humidity 55%, powdered drug was extracted with hot water and green fodder and water was available as labatum). The filtered while hot. On cooling, it was extracted with protocol was approved by Ethical Committee of carbon tetrachloride. The carbon tetrachloride was Department of Pharmacy Bahauddin Zakariya Univer- separated and washed with water and shaken with sity Multan. Groups were control (non-diabetic), control dilute ammonia solution (pink to cherry-red color in group (received vehicle only, diabetic), alloxan induced ammonia layer indicate the presence of free anthrax- diabetic rabbits treated with C. lancifolius 100 mg/kg, quinones). alloxan induced diabetic rabbits treated with C. Modified Borntrager’s test for bound anthraquinones: 5 lancifolius 200 mg/kg, and alloxan-induced diabetic g of powdered drug was extracted with ferric chloride rabbits treated with diamicron 80 mg/kg ( Hader et al., solution and hydrochloric acid. It was heated on water 1994 ). bath for 10 min and filtered while hot. On cooling, it Induction of diabetes: Animals were devided into five was extracted with carbon tetrachloride. The carbon experimental groups (control, diabetic, diabetic treated tetrachloride was separated and washed with water with 100 mg/kg body weight C. lancifolius diabetic and shaken with dilute ammonia solution color (intense treated with 200 mg/kg body weight, C. lancifolius and pink to cherry-red coloration of ammonia layer indicate diabetic treated with diamicron 80 mg/kg) Each group the presence of anthraquinones glycosides). contained six rabbits. Before starting experiment the Shinoda test for flavonoids: 5 g of drug was extracted animals in latter four groups were injected. 150 mg/kg with methanol. To methanol extract, a piece of of alloxan (sigma chemical co,) dissolved in 10% magnesium ribbon and 1 ml of concentrated isotonic saline to induce diabetes ( Issa et al., 2000 ). The hydrochloric acid were added. (Pink red or red alloxan is selective beta-cells cytotoxic drug. It coloration of the solution indicate the presence of produced all signs and symptoms of diabetes. i.e. flavonoids in the drug). hyperglycemia, glycosuria, polydipsia, polyurea, etc. The control group was injected the same volume of In vitro α-glucosidase inhibition assay: The α-glucosidase isotonic solution as that of diabetic received. The blood inhibitory activity was assessed by the standard glucose level of surviving rabbits was checked after method ( Dong et al., 2012 ) with slight modifications. three days of Alloxan injection and rabbits with blood Briefly, a volume of 60 µL of sample solution and 50 µL glucose level more than 250 mg/dL on 12 hours fasting of 0.1 M phosphate buffer (pH 6.8) containing α- were considered as diabetics ( Fridewald et
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