ABSTRACT. Extracts containing gymnemic acids, which were extracted from the leaves of Gymnema sylvestre (GS) as nine fractions, were evaluated for their effects on a high K + -induced contraction of guinea-pig ileal longitudinal muscles, on glucose transport mediated by the difference of glucose-evoked transmural potential difference (∆PD) in the inverted intestine of guinea-pig and rat, and on blood glucose in rat. Among nine fractions obtained by high performance liquid chromatography from the extract, f-2 and f-4 strongly suppressed the high K + -induced contraction of the ileal muscle, f-3 and f-5 did so moderately, and f-8 and f-9 did so weakly, whereas the other fractions did not affect it. The degree of suppression of high K + -induced contraction by f-2 at 74% was almost the same as that of f-4 at 67%, at concentrations of 0.1 mg/ml. The suppressed contraction by f-2 or f-4 was recovered by adding 5.5 mM pyruvate. The ∆PD increased by 5.5 mM glucose in the inverted intestines of guinea-pig and rat were equally suppressed by 0.1 mg/ml of f-2 or f-4 to 40%. In a rat sucrose tolerance test, f-2 and f-4 suppressed the elevation of blood glucose level. Both f-2 and f-4 suppressed the contraction of guineapig ileal longitudinal muscle, interfered with the increase in ∆PD induced by glucose in the inverted intestines of guinea-pig and rat, and inhibited the elevation of blood glucose level. In conclusion, it is suggested that some of the extracts containing gymnemic acids from GS leaves suppress the elevation of blood glucose level by inhibiting glucose uptake in the intestine. -KEY WORDS: blood glucose, gymnema extract, intestine, muscle contraction, potential difference.J. Vet. Med. Sci. 59(4): 245-251, 1997
MATERIALS AND METHODSMethod of extraction and refinement from Gymnema sylvestre leaves: Gymnema sylvestre (GS), growing naturally in South-east Asia, was extracted by a method for ordinary glycosides. GS leaves were dried and crushed, and then treated with citric acid. The treated leaves were extracted with 75% ethanol, and then the extract was evaporated. The dried extract was mixed with n-butanol and water (2:1), then the layer of n-butanol was evaporated under vacuum. The residue was washed with petroleum ether to remove fatty components, then extracted with methanol. After filtration, the methanol solution was concentrated under vacuum. The concentrated extract in methanol was separated as nine fractions by high performance liquid chromatography (HPLC) (Fig.1). The yield of the extracts from the dried leaves of GS was 0.6 g/kg.Measurement of muscle tension: Male Hartley guineapigs (Funabashi Farm, Funabashi) weighing 350 to 450g were killed by a blow to the head and exsanguinated. After exsanguination, the abdomen was opened, and the ileum was removed. The lower part of the ileum was placed in an incubation medium, and food residues were carefully removed with physiological salt solution (PSS). PSS employed was a modified Tyrode solution of the following composition (mM): NaCl, 136.8; KCl, 5.4; CaCl 2...