Öz
Suppressing the production of glucose by inhibiting α-amylase / α-glucosidase activity is one of the therapeutic approaches for decreasing postprandial hyperglycaemia and a strategy for evaluating antidiabetic activity. We investigated leaves of Spondias mombin because our previous ethnobotanical survey showed that it is used by traditional healers to manage diabetes in South West Nigeria. We report a bioactivity-guided study of S. mombin using glucose loading (1g/kg) alloxan-induced diabetic rats and inhibition of α-amylase as basis for isolation of active constituents. Hyperglycaemia was induced in albino rats and blood glucose levels monitored for 180 mins using a glucometer. Powdered leaves were macerated with 80% Methanol. The active extract was fractionated on column chromatography packed with silica gel G6OA eluting with gradient mixtures of pet. ether and ethylacetate. The most active αamylase inhibiting fraction was purified on thin layer chromatography (TLC) and pure compound identified by spectroscopy. Peak decrease in blood glucose of 41.4% (p< 0.05) was recorded after 60 mins. This activity-guided study produced an active TLC band (69.8% amylase inhibition, p < 0.05) from which β-sitosterol was characterized as the main inhibitor. This is first report of hypoglycaemic and amylase inhibitory activities of S. mombin. The role of phytosterols in control of diabetes mellitus is discussed. This study justifies the ethnopharmacological use of this species in recipes for management of diabetes mellitus.
Keywords: Spondias mombin, α-amylase inhibition, alloxan-induced diabetes, hypoglycaemic activity, hyperglycaemia
Résumé
La production du glucose en inhibant l’activité de l’enzyme α amylase / α glucosidase est l un des approche thérapeutique pour réduire l’hyperglycémie post prandiale et une stratégie pour évaluer
l’activité antidiabétique. Nous investiguons les feuilles de spondias mombin parce que notre étude ethnobotanique antérieure montrait que il était utilise par les guérisseurs traditionnels pour ménager le diabète au Sud-ouest du Nigeria. Nous rapportons l’étude de la bio activité guidée du S. mombin utilisant la concentration du glucose (1g/kg) d’alloxan induit diabète aux rats et inhibition de l’amylase comme base de l’isolation des ingrédients actif. L’hyperglycémie était induite aux rats et l’évaluation du taux du glucose pour 180 mins avec un glucomètre. Les poudres des feuilles étaient macérés avec le méthanol de 80%. L’extrait actif était fractionné a l’aide de la chromatographie a colonne rempli du gel de silicaG60A avec un gradient de mélange au pet éther et l’acide ethylacetique. La fraction du & amylase la plus inhibée était purifié la chromatographie a couche légère et le compose pur identifier par la spectroscopie. La réduction du taux du glucose était a 41.4% (P<0.05%) enregistre après 60 mins dont le beta sitostérol était le inhibiteur principal. C’est le premier rapport d’un hypoglycémiant et des activités inhibitrices de l’amylase du S. mombin. Le rôle des phytosteroides dans le contrôle du diabète mellite est discute. Cette étude justifie l’usage de l’ethnopharmacologie de ces recipes pour les soins du diabète mellite.
Correspondence: Dr. K.A. Abo, Department of Pharmacognosy, Faculty of Pharmacy, University of Ibadan, Ibadan, Nigeria. E-mail: kioabo@yahoo.com
Referanslar
Aminot-Gilchrist DV and Anderson HD. Insulin resistance-associated cardiovascular disease: potential benefits of conjugaged linoleic acid. Am. J. Clin. Nutr. 2004; 79: 1159-1163.
Wild S, Rolic G and King H. Global burden of diabetes. Diabetes Care 2004; 27(5): 1047-1053.
Tiwari A and Rao M. Diabetes mellitus and multiple therapeutic approaches of phytochemicals: Present status and future prospects. Curr. Sci. 2002; 83: 30-38.
Keay RWJ. Trees of Nigeria. Oxford: Clarendon press. 1989
Abo KA, Ogunleye VO and Ashidi JS. Antimicrobial potential of Spondias mombin, Croton zambesicus and Zygotritonia crocea. Phytother. Res. 1999; 13: 494-497.
Ashidi JS, Gible ZO and Ayodele AE. Ethnobotanical studies of anti-tuberculosis plants in Egbado, Ogun State, Nigeria Nig. J.Sci. 1997; 31.
Abo KA and Fred-Jaiyesimi AA. Ethnobotanical studies of medicinal plants used in the management of diabetes mellitus in South West Nigeria. J. Ethnopharmacol. 2008; 115(1): 67-71.
Abo KA and Jimoh TB. Anti-hyperglycaemic potential of stem bark of Bauhinia monandra Kurz in rats. Nig. J. Nat. Prod and Med. 2004; 8: 37-40.
Fred-Jaiyesimi A and Abo K. Antidiabetic activity of Gladiolus psitascinus in alloxan induced diabetic rats. Afri. J. Trad. CAM 2008; 5(2): 151-157.
Saleem R, Ahmad M, Hussain SA, et al. Hypotensive, hypoglycaemic and toxicological studies on the flavonol C-glycoside shamimin from Bombax ceiba Planta Medica 1999; 65(4): 331-334.
Harrison MA and Rae IF. General techniques of cell culture. Cambridge: University Pres, 1997, pg. 40.
Miller GL. Use of dinitrosalicylic acid reagent for the determination of reducing sugar. Anal. Chem. 1959; 31: 426.
Fred-Jaiyesimi AA. In vitro and in vivo antidiabetic activity of Parkia biglobosa and Spondias mombin. Ibadan: Ph.D Thesis, University of Ibadan. 2007, pg. 98-100.
Kim E and Goldberg M. Serum cholesterol assay using a stable Liebermann-Burchard reagent. Clin Chem. 1969; 15: 1171-1179.
Weichselbaum TE. An accurate and rapid method for the determination of protein in small amounts of blood, serum and plasma. Am. J. Clin. Path. 1946; 7: 40-49.
Reinhold JG. Standard methods of clinical chemistry. New York: Academic Press. 1953, 188.
Xin Z, Philippe G, Michel M, et al. Gram-scale chromatographic purification of β-sitosterol. Synthesis and characterization of β -sitosterol oxides. Steroids 2005; 70(13): 886-895.
Miyazawa M, Yagi N and Taguchi K. Inhibitory compounds of α-glucosidase activity from Arctium lappa L. J. Oleo Sci. 2005; 54(11): 589-594.
De Feo V and Senatore F. Medicinal plants and phytotherapy in the Amalifan coast, Salemo province, Southern Italy. J. Ethnopharmacol. 1993; 39(1): 39-51.
Adjanohoun E, Ahyi MRA, Ake AH, et al. Traditional medicine and Pharmacopoeia. Lagos: OAU/STRC. 1991.
Ashidi JS, Abo KA and Ayodele AE. Ethnobotanical survey of anti-diabetic plants in some local government areas in Ogun State, Nigeria. Afri. J. Pure & Appl. Sciences. 2005; 1(1): 6-9.
WHO, Diabetes mellitus. Second Technical Report series. Geneva: series 646.
Fathaiya J, Sihaila M and Lajis MN. Hypoglycaemic effect of stigmast– 4–en–3–one from Parkia speciosa empty pods. Food. Chem. 1995; 54: 9-13.
Kotowaroo MI, Mahomoodally MF, Gruib-Fakim A and Subratty AH. Screening of traditional anti-diabetic medicinal plants of Mauritius for possible alpha-amylase inhibitory effects in vitro. Phytother. Res. 2006; 20(3): 228-231.
Ambike SH and Rao MR. Studies on phytosterolin from the bark of Ficus religiosa. Ind. J. Pharm. 1967; 29: 91-92.
Sucrow W. Uber Stering glucoside und ein neues stigmastadienol aus Momordica charantia Tetrahedron Lett. 1965; 26: 2217.
Virdi J, Sivakami S, Shahani S, Suthar AC, Banavalika MM and Biyani MK. Antihyperglycaemic effects of three extracts from Momordica charantia. J. Ethnopharmacol. 2003; 88(1): 107-111.
Olaniyi AA, A neutral constituent of Momordica foetida. LLoydia 1975; 38: 361-362.
Marles RJ and Farnsworth NR. Antidiabetic plants and their active constituents. Phytomedicine 1995; 2(2): 137-189.
Fathaiya J, Suhaila M, Lajis MN. Hypoglycaemic effect of Parkia speciosa seeds due to the synergistic action of β-sitosterol and stigmasterol. Food chem. 1994; 49: 339-345.
Alexander-Lindo RL, Morrison EY and Nair MG. Hypoglycaemic effect of stigmast – 4- en – 3 – one and its corresponding alcohol from the bark of Anarcadium occidentale. Phytother. Res. 2004; 18(5): 403-407.
Puls W and Keup U. Influence of an alpha-amylase inhibitor (BAY d7791) on blood glucose, serum insulin and NEFA in starch loading tests in rats, dogs and man. Diabetologia 1973; 9: 97-101.