| Citrus changshan-huyou is the unique citrus specie of China with special flavor and high nutrition. It was said that the diabetes who consumed a certain quantity of Citrus changshan-huyou didn’t elevate their blood sugar level, but there was no scientific evidence. In this research, the balancing blood sugar effect of Citrus changshan-huyou and action principle were preliminarily studied.The sugar and flavonoids content in Citrus changshan-huyou pulp were determined by high performance liquid chromatography assay (dry weight):total sugar520.07±23.70mg/g, glucose94.05±5.28mg/g, sucrose308.38±10.02mg/g, fructose116.80±4.12mg/g, total flavonoids11.03±0.13mg/g, hesperidin2.23±0.09mg/g, naringin2.79±0.02mg/g. The result showed that Citrus changshan-huyou contained a lot of sugar which would elevate the blood sugar and was rich in flavonoids.The anti-diabetic effect of Citrus changshan-huyou was evaluated by animal experiments. The effect of different dose of Citrus changshan-huyou pulp on fasting blood sugar, glycogen content, the oxidation index of liver (superoxide dismutase (SOD) activity, glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) content), as well as the growth and organ index of diabetic mice were studied. The results showed that though Citrus changshan-huyou contained a lot of sugar, low and median dose (<4g/kg per day) of Citrus changshan-huyou reduced fasting blood sugar and improved the impaired oral glucose tolerance in diabetic mice. All Citrus changshan-huyou groups (<8g/kg per day) increased glycogen levels and decreased liver/body weight ratio and the accumulation of fat in the liver of diabetic mice. All Citrus changshan-huyou groups(<8g/kg per day) increased SOD and GSH-Px activity, low and median dose groups(<4g/kg per day) returned the SOD activity to normal level, low dose group(<2g/kg per day) returned the GSH-Px activity to normal level and reduced MDA content. Animal experiments results showed a moderate amount of Citrus changshan-huyou could balance blood sugar, increase glycogen and improve the oxidative stress of liver in diabetic mice.Citrus changshan-huyou pulp was partitioned into five fractions as petroleum ether, methylene dichloride, ethyl acetate, n-butanol and water fraction. The influence of the five fractions on a-glucosidase activity was studied by a-glucosidase inhibitor model. The results showed that the n-butanol fraction inhibited the activity of a-glucosidase at the same concentration. Kinetic analysis showed that the inhibitory mechanism of n-butanol fraction on a-glucosidase was competitive inhibition, and the half-inhibition concentration (IC50) was25.42±0.45mg/ml.The glucose uptake assay of3T3-L1cells showed that methylene dichloride, water fractions (>10mg/L) and ethyl acetate fraction (100mg/L) increased the glucose uptake in3T3-L1adipocytes. The n-butanol, water fraction, hesperidin (>1mg/L), methylene dichloride and ethyl acetate fraction (>10mg/L) significantly reduced the interleukin-6(IL-6) level in3T3-L1adipocytes. The methylene dichloride, ethyl acetate, water fraction (>10mg/L) and naringin (100mg/L) significantly reduced the free fatty acid (FFA) level in3T3-L1adipocytes. The results showed that there were some components could promote glucose uptake and reduce inflammatory cytokines level in adipocytes.In this research, the balancing blood sugar and increasing glycogen storage and improving liver oxidative stress effect of Citrus changshan-huyou on diabtetic mice was confirmed. The diabetic can eat Citrus changshan-huyou in a right amount(one to two for an adult per day).This effect of Citrus changshan-huyou may be associated with that it can promote glucose uptake in adipocytes and reduce the IL-6and FFA level in adipocytes and so on. |
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