Study On Extraction And Purification Of Polysaccharides From Cichorium Intybus L. And Its Immunomodulatory Effect

Cichorium intybus L.is a valuable medicinal and food homologous plant,which has higher application value,research and development prospect in the fields of health food,medicine and natural feed additives.But Cichorium intybus L.has not been fully developed in recent years.Cichorium intybus L.in Xinjiang distributes widely,and the resource is abundant.It has great potential to be processed deeply.The process conditions of extracting and purifying chicory polysaccharides(CP)from the roots of Cichorium intybus L.were studied by single factor combined with orthogonal experiment.The immunomodulatory effects of chicory polysaccharides were studied respectively on normal and cyclophosphamide(Cy)-induced immunosuppressive mice,which laid a foundation for further study of Cichorium intybus L,thus promoted the deep development and effective utilization of Cichorium intybus L.The main contents are as follows:(1)Determination of content by UV-Vis spectrophotometry(UV),total chicory sugar was determined at 492.5 nm by phenol-sulfuric acid method,and achicory reducing sugar was measured at 475.5 nm by 3,5-dinitrosalicylic acid(DNS)chromogenic method.The content of chicory polysaccharide was calculated from the difference between the two results.Both methods are simple and easy to implement with good precision,repeatability and high accuracy.(2)The extraction process of chicory polysaccharide was initially investigated by single-factor test.Then,the optimum extraction process was determined by orthogonal test:using water extraction,the amount of distilled water was 5 times that of chicory roots,90? extraction 2 times,1 h each time.The results showed that 18.93 g of chicory crude polysaccharide could be extracted from every 100 g of chicory roots.(3)Concentration volume,volume ratio of chloroform to n-butanol,volume ratio of polysaccharide solution to Sevag reagent and times of deproteinization on protein removal were studied by single factor experiment.The optimum deproteinization method was determined by orthogonal experiment:chicory polysaccharide extract concentrated to 1/8 of the original volume,chloroform:n-butanol(v/v)4:1,polysaccharide concentrate:Sevag reagent(v/v)3:1,deproteinization 3 times.The results showed that the loss rate of polysaccharides was 9.36%,and the clearance rate of protein was 49.41%.(4)In order to study the immunomodulatory effects of chicory polysaccharide on normal mice,mice were randomly divided into five groups:blank control group,positive control lentinus(LNT)group 15 mg/kg,low dose of chicory polysaccharide(75 mg/kg),middle dose(150 mg/kg),high dose(300 mg/kg).Compared with the blank group,the spleen indexes of mice administered middle and high dose of chicory polysaccharide were significantly increased(P<0.01).Furthermore,the thymus and phagocytosis indexes of mice administered middle dose of chicory polysaccharide were increased(P<0.05).Meanwhile,in order to study the immunomodulatory effect of chicory polysaccharide on immunosuppressive mice,the immunosuppressive mice model was established with cyclophosphamide(Cy).The immunosuppressive mice were randomly divided into seven groups:normal control group,model group 80 mg/kg,LNT 15 mg/kg,levamisole hydrochloride(LMS)22.5 mg/kg,low dose of chicory polysaccharide(75 mg/kg),middle dose(150 mg/kg),high dose(300 mg/kg).As a result,the spleen,thymus index and phagocytosis indexes of the normal group were higher than those of the model group(P<0.01),which indicated the successful model.Compared with the model group,in the positive control group(LNT,LMS),chicory polysaccharide low,middle and high dose groups,the spleen indexes of immunosuppressed mice increased by 48.61%,68.42%,59.17%,77.98%,77.23%(P<0.01);the thymus index was increased by 23.06%(P<0.05),16.53%,15.45%,31.18%(P<0.01),24.58%(P<0.05);the phagocytosis index increased by 10.27%(P<0.05),5.02%,8.12%,10.36%,20.99%(P<0.01).The results showed that chicory polysaccharides have a certain effect on regulating immunity.