Premilinary Study On The Hypolipemic Mechanism And Affinity Chromatography Of Polysaccharides From Camellia Chrysantha (Hu) Tuyama

Camellia chrysantha(Hu)Tuyama is a kind of plant which is in theaceae ?camellia ? camellia genus.It is called “the queen of the camellia's world” and “the ailuropoda of the plant's domain” because it has flavous petal which is unique in the camellia.The polysaccharide from leaves of Camellia chrysantha(Hu)Tuyama is one of the active ingredients from Camellia chrysantha(Hu)Tuyama.It is reported that the polysaccharide from Camellia chrysantha(Hu)Tuyama has obvious hypolipidemia activity.But how the polysaccharide from Camellia chrysantha(Hu)Tuyama depressing blood-fat is not clear.In addition,though ion exchanged chromatographic column has been used to purify Camellia chrysantha polysaccharide,but it is not a labour-saving way.In this article,on the one hand,we preliminary researched the way of Camellia chrysantha polysaccharide depressing blood-fat;on the other hand,we researched the the agglutination reaction about the polysaccharide from leaves of Camellia chrysantha(Hu)Tuyama with ricinus communis agglutinin I and other lectins,found the appropriate lectin as the ligand of affinity absorbent,made the affinity absorbents and purified Camellia chrysantha polysaccharide,and preliminary researched the physicochemical property of purified part.The main contents and conclusions were listed as followed:1?Preliminary study on the mechanism of pressing blood-fat of Camellia chrysantha polysaccharide.The effect of Camellia chrysantha polysaccharide on the activity of3-hydroxy-3-methyl-glutaryl coenzyme A reductase(HMGCR)? fatty acid synthetase(FAS)? NADP-malate dehydrogenase(NADP-MDH)? glucose-6-phosphate dehydrogenase(G-6-PDH)was studied.The results showed that camellia chrysantha polysaccharide can,t significantly inhibit HMGCR activity;can significantly inhibit FAS activity;can significantly improve the activity of NADP-MDH and G-6-PDH.So it was speculated that Camellia chrysantha polysaccharide can't inhibit the synthesis of cholesterol by inhibiting HMGCR activity;may inhibit the synthesis of fatty acid by significantly inhibiting FAS activity;can't inhibit the synthesis of cholesterol and fatty acid by inhibiting NADP-MDH and G-6-PDH activity.2?Preliminary study on agglutination reaction characteristics of polysaccharide fromleaves of Camellia chrysantha(Hu)Tuyama.The agglutination reaction of the polysaccharide from leaves of Camellia chrysantha(Hu)Tuyama with ricinus communis agglutinin I,cristagalli Lectin,euonymus europaeus lectin,maackia amurensis lectin,concanavalin A and peanut lectin,were discussed respectively by colorimetric method.The results showed that the agglutination can react with all of the lectins,but the reaction degree were remarkable different.The degree of agglutination reaction with ricinus communis agglutinin I was the strongest,where galactose was the specific binding sugar residues.In contrast,the degree of agglutination with cristagalli Lectin,euonymus europaeus lectin,maackia amurensis lectin and peanut lectin,were rather weak,although their specific binding sugar residue were similar.The absorbance of the agglutination reaction of ricinus communis agglutinin I was 6~10 times related to that of other lectins,while the polysaccharide concent ration increasing to 6.59 mg/mL.3?The preparation of ricinus agglutinin affinity chromatography and the optimization of chromatographic conditions.Using ricinus agglutinin as ligand,affinity absorbent RCA-Sepharose 2B and RCA-Amino were mad by respectively coupling ricinus agglutinin with Sepharose-2B and AF-Amino-650 M,and their performance were compered.The results showed that the coupling ratio of lectin with AF-Amino-650 M and Sepharose 2B respectively were 91% and 71%;unde the same conditions,The affinity rates of Camellia chrysantha polysaccharide with RCA-Sepharose 2B and RCA-Amino respectively were 21.4 and 25.3%;in addition,the flow speed of mobile phase through the RCA-Amino affinity column drop rapidly with the increase of frequency of use.Through comprehensive consideration,RCA-Sepharose 2B was selected and used for purifying Camellia chrysantha polysaccharide.Optimization of the conditions of chromatography.The effect of adding sample quantity on the affinity rate of Camellia chrysantha polysaccharide with RCA-Sepharose2 B ? the reviving effect of RCA-Sepharose 2B by phosphate buffer with different concentration of sodium chloride and the volume of phosphate buffer were respectively researched.It showed that the affinity rate of Camellia chrysantha polysaccharide with RCA-Sepharose 2B was biggest(57%)when the adding sample quantity was 10.4 mg/g;the reviving effect was best when affinity column was washing by no less than 15 times of column volume of phosphate buffer with 0.2M of sodium chloride.4?Preliminary analysis of physicochemical of affinity separation polysaccharide.The molecular weight and constitute of polysaccharide were respectively studied by high performance gel permeation chromatography and gas chromatography.It showed that there are significant differences between chromatogram of crude camellia chrysanthapolysaccharide and its purified components.Though purified components was still mixture,but it is sure that crude polysaccharide was purified to a certain extent.Camellia chrysantha polysaccharide and bound fraction was at least composed of rhamnose?arabinose?xylose and galactose.The proportional of rhamnose?arabinose?xylose and other monosaccharide in Camellia chrysantha polysaccharide was 5.06%?44.36%?5.13%?45.5%.The proportional of rhamnose?arabinose?xylose and other monosaccharide in bound fraction was 5.78%?47.5%?7.80%?38.9%.