A Comparison Of The Immunostimulatory Of The Polysaccharide From Tetraploid And Diploid Echinacea Purpurea

Echinacea is one of important medicinal herbs with dual effect of immunopotentiating agents and anti-inflammation,therefore it has good prospects of application as a raw material of medicinal plant.Although Echinacea has been introduced and cultivated in many countries and regions around the world,and literature about polyploid study are rarely reported.We used biotechnological method to obtain the character of biological stableness of tetraploid E.Purpurea.Previous studied in our laboratory showed the total fresh root weight and total dry root weight of tetraploid Echinacea were higher than those of diploid plants and also revealed that tetraploid E.purpurea plants had higher contents of alkamides and caffeic acid derivatives than diploid E.purpurea plants.Regrettably,the effects of Echinacea ploidy levels on polysaccharide content and immune modulation remain unknown.In this study,we compared the content and composition of polysaccharide of diploid and tetraplo id E.purpurea.We investigated the methods of MTT and ELISA to detect lymphocyte proliferation and measure the levels of IL-2,IFN-? and TNF-?in normal mice and immunosuppressed mice model that were made by cyclophosphamide in vitro and in vivo,Comparison of the immunomodulatory effects of crude diploidE.purpurea Polysaccharides and crude tetraploid E.purpurea Polysaccharides.Main results and conclusions are summarized as follows:1.The total polysaccharide content of the medicinal ingredients in crude tetraploid E.purpurea polysaccharidewas higher than that in crude diploid E.purpureapolysaccharide.The polysaccharide content of the extraction was measured with the optimized way of coloration of phenol-sulfuric acid method,results show that the content of polysaccharide in diploid and tetraploid E.purpureais 44.65% and 85.51% respectively.2.The results of 1H-NMR spectra and GPC chromatogram showed that there were no differences between the diploid and tetraploid E.purpurea polysaccharides in the overall structure and molecular weight.All the results indicated that the chromosome doubling did not lead to the synthesis of new E.purpurea polysaccharides,but increased the amount of the original accumulation of polysaccharides.3.In vitro,results showed that crude diploid E.purpurea polysaccharide(EP2)and crude tetraploid E.purpurea polysaccharide(EP4)could promote the proliferation and secretion of IL-2 and IFN-? of normal mouse spleen lymphocytes stimulated by ConA.On the other hand,the EP4 at low concentrations(0.0312 mg/mL)were able to maximally stimulate lymphocyte proliferation and secretion of IL-2 and IFN-? of normal mouse spleen lymphocytes stimulated by ConA.While mouse spleen lymphocyte stimulation and secretion of IL-2 and IFN-? were the best when the EP2 concentration was 0.125 mg/mL.4.In vivo,results showed that EP4 and EP2 can significantly improve the activation and proliferation of lymphocytes and secreted IL-2,IFN-? and TNF-? in immune function mice induced by cyclophosphamide.EP4 was more effective in promoting the proliferation of lymphocyte and secreted IL-2,IFN-? and TNF-? in mice with immunosuppressed by cyclophosphamide than EP2 in the same concentration.Taken together,these data demonstrated that polysaccharide content increased with the doubling in chromosomal number,but the chromosome doubling did not lead to the synthesis of new E.purpurea polysaccharides.EP2 and EP4 was more effective in promot ing the proliferation of lymphocyte and secretion of IL-2,IFN-? and IFN-? in normal mice and mice with immunosuppressed by cyclophosphamide,EP4 effect of immunization was significantly higher than EP2,tetraploid E.purpureahad stronger immune activities.