The Effect Of Cyclocarya Paliurus(Batal.) Iljinskaja Polysaccharide On Cell Proliferation And Lipid Metabolism In3T3-L1Adipocyte

This study was conducted to investigate the effect of cyclocarya paliuruspolysaccharide. The crude extraction was isolated and purified by D301-Rmacroporous resin and DEAE, and there were four different components. Then usedGel Permeation Chromatography and Multi-angle Laser Light Scattering to determinethe molecular weight.3T3-L1preadipocyte cells were used and treated with differentdoes of cyclocarya paliurus polysaccharide to detect its hypolipemic effect. ThemRNA expression of PPARγ、PPARα、PPARδ (peroxidase proliferation of activatedreceptors)、C/EBP-α (CCAAT/enhancer binding protein-α)、FAS (fatty acid synthase)and Glut-4(glucose transporter-4) on cells treated with different does of differentcomponent of cyclocarya paliurus polysaccharide were determined by Q-PCR, theprotein expression of related genes were investigated by western-blot. Theexperimental results showed that:(1) The crude extraction was isolated and purified by D301-R macroporousresin and DEAE, and there were four different components. Then use GelPermeation Chromatography and Multi-angle Laser Light Scattering todetermine the molecular weight.The molecular weight were39640Da,2098Da,64530Da,4236Da,1357Da,46570Da,4443Da,1334Da,3915Da,1479Da.(2) In the MTT experiment, the cyclocarya paliurus polysaccharide had littletoxicity on3T3-L1preadipocyte cells. The polysaccharide could reducethe consumption of glucose of3T3-L1preadipocyte cells, when treatedwith the component Ⅲ of cyclocarya paliurus polysaccharide at theconcentration of150μg/ml, the inhibition ratio was9.61%. When treatedwith the component Ⅲ of cyclocarya paliurus polysaccharide at theconcentration of120μg/ml, the extracellular glycerol content wasdebased by30.8%.(3) From the examineion of the effect of cyclocarya paliurus polysaccharideon the expression of specifical genes during adipocyte differentiation, astreated time prolong, the expression of PPARδ gene was notable ascenged,and the expression of PPARγ、PPARα、C/EBPα、Glut-4and FAS geneswere notable descenged, compared with the control group.(4) As the treated time prolong, the expression of PPARα protein of theexperimental group in3T3-L1adipocyte cells was increased after treatedwith component Ⅰa nd Ⅲ ofcyclocarya paliurus polysaccharide, whiletreated with componentⅡand Ⅳ of cyclocarya paliurus polysaccharide,the expression of PPARα protein was reduced.The expression of FASprotein was significantly improved after the3T3-L1adipocyte cells wastreated with component Ⅰ, Ⅱand Ⅳ of cyclocarya paliuruspolysaccharide, however, when treated with the component Ⅲ ofcyclocarya paliurus polysaccharide at the concentration of90μg/ml and120μg/ml, the expression of FAS protein were significantly improved orno statistical difference, and the expression of FAS protein weresignificantly dropped or no statistical difference when the concentrationwas150μg/ml. The expression of Glut-4protein was significantly reducedafter the3T3-L1adipocyte cells was treared componentⅠ, Ⅲ and Ⅳ ofcyclocarya paliurus polysaccharide,while when treated with the component Ⅱ of cyclocarya paliurus polysaccharide at the concentrationof30μg/ml and90μg/ml, the expression of FAS protein weresignificantly improved, and the expression of FAS protein weresignificantly dropped when the concentration was60μg/ml.