Study Of Chuanmingshen Violaceum Polysaccharides Extraction Process And Its Immune Activity

1. Chuanmingshen violaceum polysaccharides extraction process optimization In order to get the best extraction process of Chuanmingshen violaceum polysaccharides and establish a reliable purification drying methods, the next two methods were used. The one, the polysaccharides which content in the extraction water as index, and the phenol-sulfuric acid method combined with DNS method were used to determination it; the other, single factor and orthogonal experiment were used to determin the best best technology of Chuanmingshen violaceum polysaccharides extraction. The best extraction process of Chuanmingshen violaceum polysaccharides were showed as following:the extraction temperature was 100℃, the extraction time was 4h, the the expected solid-liquid ratio was 1:30 and its had been Crushed into 80 mesh; the established method was a efficient, simple, low cost, and the purity of polysaccharides obtained by this method can reach 93.85%. The results may serve as basis of the extraction in large scale or for its further research.2. The effect of Chuanmingshen Violaceum Polysaccharides and its solfated derivatives on the proliferation of mice spleen lymphocytes in vitro In oder to investigate the effect of Chuanmingshen Violaceum Polysaccharides(CVP) and solfated Chuanmingshen Violaceum Polysaccharides(SCVP) on the proliferation of mice spleen lymphocytes in vitro. The Treated with ConA, the B cells treated with LPS;the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltet-razolium bromide, MTT) method was used to detect the proliferation of spleen lymphocytes of each group in vitro. When the cells were treated with ConA in the present of CVP at the concentrations of 15-375μg/mL and SCVP at each tested concentration, the lymphocytes proliferation rate were significantly (P< 0.01) higher than that of the control group; When the cells were treated with LPS in the present of SCVP at the concentrations of 15-375μg/mL,the lymphocytes proliferation rate were significantly (P< 0.01) higher than that of the control group; the lymphocytes proliferation rate was not significantly compared with the control group when the CVP were added to the resting cells alone or the cells were treated with LPS; the lymphocytes proliferation rate were significantly (P< 0.01) higher than that of the control group when the SCVP were added to the resting cells at the concentrations of 75-375μg/mL. The results showed that the CVP and SCVP can promote the proliferation of mice spleen lymphocytes in vitro, Suggesting that CVP and SCVP have the immune-enhancement activity, and can irritant the spleen lymphocytes of mice as a mitogen.3. Effect of Chuanmingshen violaceum polysaccharides and its sulfated derivatives on immunosuppression induced by cyclophosphamide in mice In oder to investigate the effect of Chuanmingshen Violaceum Polysaccharides (CVP) and solfated Chuanmingshen violaceum polysaccharides (SCVP) on immunosuppression induced by cyclophosphamide (CY) in mice. CY were used to induce immunosuppression in mice; Spleen and thymus indexes were used to evaluate the immune organs indexes; the (3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltet-razolium bromide, MTT) method was used to detect the proliferation of spleen lymphocytes of each group; the concentrations of IFN-y and IL-2 were assayed by ELISA kit. SCVP and CVP could resist immunosuppression by promoting lymphocyte proliferation, increasing the contents of IFN-y and IL-2, promoting immune organs development in immunosuppressive mice induced by CY. SCVP and CVP exhibited the potential to used as immunopotentiator.4. Effects of Polysaccharide from Chuanminshen violaceum on Immune Response of Newcastle Disease Vaccine in Chicken In oder to investigate the effect of Chuanmingshen Violaceum Polysaccharides (CVP) on Immune Response of Newcastle Disease Vaccine in Chicken.180 three-yellow chickens at one day of age were randomly divided into four groups and reared in separated pens. On 7 days old, the average maternal serum hemaglutination inhibition (HI) antibody titer was less than 3 log2, all chickens of each group (the average body weight (BW) was 120 g) were vaccinated with ND live vaccine through nasal drip and eye-drop. At the same time, the chickens in three CVP groups (high, medium and low doses of CVP) were orally administered with 0.5 mL of CVP at concentrations of 100 mg/kg BW,50 mg/kg BW,25 mg/kg BW respectively, once a day for fi ve successive day, in negative control group (NC), with 0.5 mL of physiological saline, once a day for fi ve successive day. On days 14,21 and 28, the serum antibody titer, erythrocyte-C3b receptor rosette rate (E-C3bRR), erythrocyte-C3b immune complex rosette rate (E-ICRR), peripheral lymphocyte proliferation and peripheral CD4+/CD8+ratio were measured. The results showed that the antibody titer, E-C3bRR, elimination rate of immune complex and peripheral lymphocyte proliferation in three CVP groups and peripheral CD4+/CD8+ ratio in medium dosage of CVP group were signifi cantly higher (P< 0.05) than those in control group throughout the process of whole experiment almost. The CVP not only improved the E-C3bRR and accelerated the elimination rate of CIC, but also induced higher antibody titer, peripheral lymphocyte proliferation and peripheral CD4+ /CD8+ ratio in chickens vaccinated against ND live vaccine. This indicated that CVP possessed immune-enhancement effi cacy of ND live vaccine and might be expected as a candidate of new-type adjuvant.