Abstract
1,5-Anhydro-D-fructose (AF) is a mono-saccharide directly formed from starch and glycogen by the action of α-1,4-glucan lyase (EC 4.2.2.13). Our previous study has indicated that AF increases glucose tolerance and insulin secretion in NMRI mice after administration through a gastric gavage in a single dose at 150 mg per mouse. In this study, we used high-fat feeding of C57BL/6J mice to examine the influence of long-term administration of AF on glucose-stimulated insulin secretion in vivo and in vitro. We found that 8-weeks of high-fat feeding increased body weight, fasting blood glucose and insulin levels in C57BL/6J mice when compared to mice fed normal diet. Impaired glucose tolerance was also observed in mice receiving 8-weeks of high-fat diet. In contrast, AF (1.5 g/kg/day), administered through drinking water for 8-weeks, did not affect body weight or food and water intake in mice fed either the high-fat or normal diet. There was no difference in basal blood glucose or insulin levels between AF-treated and control group. Oral glucose tolerance test (OGTT) showed that AF did not affect glucose-stimulated insulin secretion in mice. In in vitro studies with isolated islets, AF did not influence glucose-stimulated insulin secretion in mice receiving either high-fat or normal diet. We therefore conclude that when given through drinking water for 8 weeks at 1.5 g/kg/day, AF has no effect on glucose-stimulated insulin secretion in C57BL/6J mice challenged with a high-fat diet.
Highlights
1,5-Anhydro-D-fructose (AF) is a mono-saccharide directly formed from starch and glycogen by the action of a-1,4glucan lyase (EC 4.2.2.13)
We have previously shown that when given through a gastric gavage (150 mg) together with glucose (150 mg/mouse), AF induces glucose tolerance, insulin secretion and increases in plasma levels of glucagon-like peptide-1 (GLP-1) [13]
AF treatment for 8 weeks As seen in Table 1, 8 weeks of high-fat feeding increased body weight and blood glucose compared with the mice fed normal diet
Summary
Animal Four-week old female C57BL/6J mice weighing 15 g were obtained from Bomholtgaard Breeding and Research Center, Denmark. Oral glucose tolerance test After 8 weeks of AF treatment, blood was drawn from the intra-orbital bullar plexus of all mice for the measurement of basal glucose and insulin levels. For oral glucose tolerance test (OGTT), mice fasting overnight were given glucose (150 mg/mouse) orally and their blood was collected at times 0, 15, 30, 60, 90 and 120 min following glucose administration. After washing the incubated islets for three times with HBSS, they were handpicked under a stereomicroscope and incubated overnight in RPMI 1640 medium supplemented with 10% fetal bovine serum, 2.05 mmol/l L-glutamine, 2.5 μg/ml amphotericin B, 100 IU/ml penicillin and 100 μg/ ml streptomycin at 37°C in humidified air equilibrated with 5% CO2. The islets were washed three times and pre-incubated for 60 min at 37°C in a Hepes medium (pH 7.4 supplemented with 0.1% human serum albumin (Sigma) and 3.3 mmol/l glucose. Statistical comparisons for differences between AF and control were performed by unpaired Student t test
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