The Protective Effect Of Kudingcha Polysaccharide On Liver Injury And Endothelial Dysfunction Triggered By High Fructose Diet

Polysaccharide is one of the most important component in plant, which has many biological activities, such as antioxidant, anti-inflammatory and anti-hyperlipidemia. Ilex kudingcha belongs to aquifoliaceae. It has been called gao Lou or Lou in ancient time. As we all know, it distributs widely in our courtry and has been cultivated mainly in hunan, hubei, guangdong and hainan province.This paper examined the clarified of polysaccharide from Ilex kudingcha and the antioxidant, anti-inflammatory and anti-hyperlipidemia effects, providing a theoretical foundation and technical application of Hex kudingcha polysaccharide.The polysaccharide was obtained with the water extraction and alcohol precipitation method. The total carbohydrate content and monosaccharide composition were measured by phenol-sulfuric acid method and 1-phenyl-3-methyl-5-pyrazolonepre-column derivatization HPLC method. The Ilex kudingcha polysaccharide extraction rate was 6.5% and the total carbohydrate content of Ilex kudingcha polysaccharide polysaccharide was 59.6%, which was composed of D-mannose, D-ribose, D-rhamnose, D-glucuronic acid, D-galactose acid, D-glucose, D- xylose and D- galactose with a relative molar percentage of 3.1%, 8.6%?4.1%?7%?3.4%?7.0%?31% and 24%. The results showed that Ilex kudingcha polysaccharide polysaccharide was a typical heteropolysaccharide.As compared to the model mice, the enzymatic activities of serum ALT and AST in the administration of Ilex kudingcha polysaccharide at 800 mg/kgbw decreased to 18.46 ± 0.57 U/L and 58.42 ± 1.06 U/L from 23.72 ± 0.94 (p?0.01) and 88.05 ± 1.24 U/L(p<0.01). MDA activities decreased to 0.88 ± 0.08 nmol/mgprot (p<0.01). SOD and GSH-Px activities were increased to 22.30± 0.91 U/mg prot (p<0.01) and 1188.44± 86.97 U/mgprot (p?0.01) respectively. The mice liver with H&E staining and oil red O staining reflect that high fructose-diet makes the model group mice liver injured. There were edema and necrosis and cell morphology was different. The nucleus was atrophic and the cell has more hollow. However, the administration of Ilex kudingcha polysaccharide at 200,400 and 800 mg/kgbw could effectively attenuate this phenomenon. The fact indicated that Ilex kudingcha polysaccharide could effectively exhibited strong protective effects against high fructose-induced liver injury in mice.Comparing to the model mice, the protective administration of Ilex kudingcha polysaccharide at 800 mg/kg·bw could effectively attenuate an increase in TC, TG and LDL-C levels and a decrease in HDL-C concentrations, respectively. As expected, TNF-a and TXA2 levels decreased to 13.64 ± 1.04 pg/mL (p<0.01) and 170.24 ± 9.24 mmol/L (p<0.01) and PGI2 levels increased to 51.64 ± 1.98 mmol/L (p<0.01) at 800 mg/kg·bw. The mice vascular tissues reflect that the concentration at 800 mg/kg·bw showed near normal appearance. This suggested that high dietary fructose clearly caused vascular endothelial injury in mice and Ilex kudingcha polysaccharide could effectively improve the dysfunction of vascular endothelial system.Hex kudingcha polysaccharide show three peaks area at 0.05,0.15 and 0.3 mol/L NaCl concentration after DEAE-52 cellulose elution. The samples of 0.15M were further purified by Sepharose G-100 and the elution peaks of two peaks were separated, called KD-1 and KD-2. Infrared spectroscopy showed KD-1 and KD-2 both had the characteristic absorption peak of polysaccharide, which have pyranoid ring in KD-1 and KD-2. The SEM electron microscope reflected that the properties of KD-1 are more brittle and the properties of KD-2 is net shape. HPLC analysis results that monosaccharide composition of KD-2 was D-mannose (7.6%), D-rhamnose (12.3%), D-galacturonic acid (2.7%), D-xylose (58.4%), D-galactose (3.9%) and D-arabinose (14.6%). The highest content was D-xylose (58.4%) and D-arabinos (14.6%).