Extraction And Purification Of Schisandrae Polysaccharide And Its Effect On The Intestinal Immune Function

In recent years,a large number of studies have shown that polysaccharide has a variety ofphysiological health functions, but polysaccharide on intestinal immune function is not yet clear.In thisstudy, the dried fruits of Schisandra are used as the main raw material, using the UF method to preparehigh purity of Schisandra fine polysaccharide.The study is about Schisandra fine polysaccharide-inducedapoptosis of tumor cells and on the intestinal immune regulation of mice.The main findings are asfollows:1.Determination of the optimal preparation of Schisandra fine polysaccharides. Schisandra powderhas been in hot water at55℃for2h after defatting. Use ultrafiltration membrane to separate and purifythe extracted liquid through100ku,The best ultrafiltration pressure is0.207Mpa,the operatingtemperature is20°C,the ultrafiltration time is30min.The ultrafiltrate remove the protein,then pasteurizeand freeze-dry the ultrafiltrate,the average molecular weight of the Schisandra fine polysaccharide is210ku.2.The refined polysaccharide prepared from Schisandra was dissolved in culture medium so that thefinal concentrations were0,0.05,0.1,0.25,0.5,0.75,1.0,1.5,2mg/L and role in SMMC7721hepatoma cells,the results show that in a short time of treatment (24to48hours),Schisandra polysaccharide has nosignificant apoptosis (P>0.05) on liver cancer cells;when in24hours,the higher dose even promoteproliferation partly,in longer processing time (72h), the performance of polysaccharide has significanteffect on hepatoma cell growth inhibition and promotion of apoptosis (P<0.05),the dose size and cellviability is negatively correlated. Acridine orange,ethidium bromide staining polysaccharides processhepatoma cells for72h,blank group of cells almost rendered green, form morphological integrity of thecells and a complete adherent growth;in the group of the polysaccharide concentration0.05,0.5,2.0mg/L,the number of orange dead cells increase obviously,cell morphology and size aredisorganized,the growth of the cells attached to the edge become blurred and they appear thephenomenon of apoptosis with a positive correlation between dose,this further illustrates that theSchisandra fine polysaccharides can induce SMMC7721hepatocellular carcinoma cell apoptosis.3.Use Schisandra powdered polysaccharides of high-dose (500mg/kg d), middle-dose (200mg/kg d), low-dose (50mg/kg d) to feed BALB/c mice for15days.The SIgA in the intestinal fluid ofintestinal and the main cells factor levels of the small intestine tissue are basically sTab.le after thefluctuations of about the first7days.Immune response peak of the first seven days, IL-2, IFN-γ, IL-4,IL-10and other cytokines have been a peak successively, the intestinal mucosal immune system of themice has a cellular immune dominant,a humoral immune dominant in succession,then the immuneresponse slow down gradually.The results of long-term gavage for10~15days showed thatschisandra fine polysaccharide can promote the secretion of intestinal SIgA, SIgA content and the dosewas positively correlated.Flow cytometry analysis mesenteric lymph nodes (MLN) T-lymphocytephenotype showed that high dose and middle dose group could significantly increase the CD3+CD4+ and CD3+CD8+ratio.Combination of the cytokines and SIgA content changes before,inferred thathigh and middle doses of polysaccharide increase the proportion by increasing the number of cells inCD3+CD4+the phenotype of helper T cells of different subsets.Thereinto,the high dose group canincrease the number and activity of Th2cell subsets,the middle dose group increased the activity andnumber of Th1, Th2subsets cell,CD3+CD4+and CD3+CD8+proportion of the low-dose group wasnot significant compared with the control group (P>0.05),but they can increase the Th1, Th2cellactivity.4.Middle-dose (200mg/kg d), low-dose (50mg/kg d) Polysaccharide gavage can promoteintestinal mucosal immunity levels,improve both humoral and cellular immunity, maintain intestinalimmune balance and resist to the foreign matter infection,.so that it is favorable for the body intestinalhealth; high-dose group (500mg/kg d)can increase the level of intestinal humoral immune,but causedintestinal cell-mediated immunity is suppressed, the body is not sensitive to foreign matter,and it is notconducive for our body’s intestinal health...