The Extraction、Purification And The Function Of Anti-hyperlipidemia Of Polysaccharide From Enteromorpha

OBJECT： Study the way to purify and separate polysaccharide from enteromorpha. Comparethe difference between the traditional way and the high efficiency ways. To observe the effectsof EP, which was extracted by two different ways. Adjusting the lipids metabolism ofhypolipidemic Rats and discuss its possible mechanisms.METHOD:1.Take the enteromorpha which is produced in Fujian province as the research object. DEAE-cellulose and Sephadex-100will be used to purify and separate the polysaccharide ofenteromorpha(EP). IR will be used to determine the structural features of EP and GC will be usedto study the composition of monosaccharide of EP.2.Using spectrophotometer to compare the polysaccharide、 glycuronate、 sulfateradical.3. Rats model of Hypolipidemic was established by a high sugar and fat diet and thengiven various doses and kinds of Ep for40days. Observe the generalsituation,measured the daily weight; At the end of the experiment, evaluated thelevels of TC,TG,HDL,LDL,SOD and GSH-Px. Observed the liver organizationstructure by HE staining; detected the expression of LactmRNA、HmgcrmRNA andAbcA1mRNA in liver tissue by realtime-PCR.RESULT:1.The horizontal comparative among the different ways to extract EPThe proportion of active group in polysaccharide from EP is almost the same. In thesign way, enzyme can increase the extraction of volume of EP by most. The two-waysmethod can increase the extraction compare the sign method.. The three-ways methodcan increase the extraction compare the two-ways method.2. Purify、 separate and analysis of Polysaccharide From Enteromorpha EP wasseparated to4components, EP-1,EP-2,EP-3,EP-4by DEAE-cellulose. EP2takes upthe largest part,81%. EP-2shows a single symmetric peak when it flow throughSephadex-100. IR shows that EP-2is a kind of carbohydrate and contents thiol esters.GS shows that EP-2was made up of rhamnose, glucuronic acid, arabinose, fucose, xylose and glucuronic. The proportion of each monosaccharide is5.12：1.32：3.38：1.62：1：1.033. General situationDuring the experiment the general situation of all the groups are good, but at the endof the experiment the reaction and pelage of model group got worse. Compare withthe model group the situation of control group and the EP groups are better.4. blood lipid indexCompared with blank control group, TC、TG、LDL of model group were declinedsignificantly, HDL increased obviously(P<0.05);Compared with model group, TC andTG of EHEP、EMEP、ELEP、UMEP、ULEP groups were declined significantly(P<0.05); Compared with model group，HDL of ELEP and ULEP were increasedsignificantly(P<0.05); Compared with control group，TG of each EP group weredeclined significantly.5antioxidant indexCompared with blank control group, SOD and GSH-Px of model group was lowersignificantly (P<0.05); Compared with model group, SOD of each EP group andcontrol group were higher (P<0.05); Compared with model group, GSH-Px of eachEP group were higher (P<0.05)6. HE staining of liverHE staining of liver: The hepatic lobule of blank control group is normal. The modelgroup shows obscure structure of hepatic lobule and cords. Compared with modelgroup and all the EP groups, the hepatic cords of control group shows a bettercondition. There is no significant difference between the same dose EP of the twokinds of the same dose.7expression level of mRNACompared with model group, the expression of Lact in EHEP groups were higher(P<0.05),other groups have no significant different but the control group shows arising trend. There is no significant difference between each group in the expression of Hmgar on statistics, but the trend of the expression of the groups which are treatedby EP is rising. There is no significant difference between each group in theexpression of AbcA1on statistics and trend.Conclusion:1.The high efficiency way can increase the extraction of polysaccharide but have alittle efficiency in improving the extraction of glycuronate、sulfate radical.2.The major constitution of EP is EP-2.EP-2is a kind of polysaccharide whichcontents thiol esters.3. The bioactive of EP3.1EP can play a certain role in adjusting lipid metabolism in hyperlipidemia rats,enhance antioxidant effect, restrain oxidative damage, repair the damaged of liver ata suitable dose.3.2EP can make the expression of Lact and Hmgcr to a proper level to get thefunction of adjusting TC、TG then prevent hyperlipidemia.3.3The biological activity of reducing blood lipids of EP which is extracted byenzyme is a little better than the EP extracted by ultrasonic. Both EP has their ownadvantage in antioxidant.