Filtration On The Extraction Process And The Immunomodulation Effect Of The Prinsepia Utilis Royle Flavonoids In Chicken

OBJECTIVE:Filtration on the Extraction proces and the immunomodulation effect of the Prinsepia Utilis Royle flavonoids in chicken.EXPERIMENTATION 1:Filtration on the Extraction process of the Prinsepia Utilis Royle flavonoids.Methods:Choose the extrac method of Prinsepia Utilis Royl flavonoids from ethanol extraction,hot water extraction,alkali dissoltin and acid depositon.The content of ethanolsoution,temperature,volune of ethanol solution and time as four indic factor to analyse,then they were optimized by the method of L₉(3⁴)orthogonal experiment.Color reaction(HC1-Mg reaction:from yellow to red;NaBH₄ reaction:maqenta;A1Cl₃ and MgAC₂ reaction:qlaucous fluorescence under 365nm ultraviolet radiation;Zr- citric acid reaction:bright yellow,the color fade after addition 2% carbinol of citric acid;H₃BO₃ reaction:kelly and trifling fluorescence)were used for identification and the content of flavonoids was determined by the method of ultraviolet spectrophotometry method.Results:The best extraction process was like this:ethanol concentration 60%,the ration of solid to liquid 1:20 and extracting temperature 80℃for extracting time 3h.The crude extract is flavonoids component was identified by the color reaction.The content of flavonoids in the Prinsepia Utilis Royle was 5.23%.EXPERIMENTATION 2:The immunomodulation effect of the Prinsepia Utilis Royle flavonoids in chickenMethods:Sixty 1-day-old chicken were divided into three groups randomly.Groupâ… feeded with basal diet.Groupâ…¡and groupâ…¢was supplemented with 200 mg/kg and 400 mg/kg Prinsepia Utilis Royle flavonoids in the basal diet respectively.After 14d,21d and 28d the group which effect of weighting is best was picked out to blood and immune organize sampling.Blood be use to mensurate antibody level,blood Biochemical Indicator,E-C3bRR,E-ICR,Et,Ea,percent of T lymphocytes.Immune organize be used to mensurate the index of immune organ,the quantity of T lymphocyte and macrophage.And then we detected change of structure and mensurated diameter of lymph follicle after making splices and HE staining.Otherwise ultrastructure of spleen was be observed under electron microscope.Results:The exhibition which influence of Prinsepia Utilis Royle flavonoids for blood serum antibody and blood biochemical indicator in chicken was like this:compared with the contrast group the content of IgM in trial group increased distinctly at 21d(p＜0.01),the content of IgG and IgM in trial group increased significantly at 28d(p＜0.05).Others haven't significant change(p＞0.05).Each Blood Biochemical Indicator have not significant change(p＞0.05)and all change in normal range.The exhibition which influence of Prinsepia Utilis Royle flavonoids for immunity of RBC and lymphocyte in chicken was like this:compared with the contrast group E-C3bRR and E-ICR in trial group increased appreciably at all days(p＞0.05);Ea in trial group increased distinctly at all days(p＜0.01);Et in trial group increased distinctly at 21d(p＜0.01).The percent of T lymphocyte in trial group increased distinctly at 21,28d(p＜0.01).The exhibition which influence of Prinsepia Utilis Royle flavonoids for immune organ and immunocompetent cell(ICC)in chicken was like this:compared with the contrast group the index of bursa of Fabricius and thymus in trial group increased significantly at 21d(p＜0.05).The index of spleen and bursa of Fabricius in trial group increased significantly at 28d(p＜0.05);The index of thymus increased distinctly at 28d(p＜0.01).The percent of T lymphocyte in trial group increased significantly at 14,21,28d.The percent of macrophage in trial group increased significantly at 28d(p<0.05).The exhibition which influence of Prinsepia Utilis Royle flavonoids for histology of immune organ in chicken was like this:be observed by light microscope,all organization of immune was immature at 7d.Red pulp and white pulp were undivided in spleen;Rugged mucosa and sparse cell were observed in bursa of Fabricius;There were large or medium sized lymphocyte in thymus. Compared with the contrast group at 14,red pulp and white pulp were divided in spleen and more lymphocyte round central artery;The cortex and medulla begin to separate and comparatively smooth mucosa and medium sized follicle were appeared in bursa of Fabricius;There was clear borderline between cortex and medulla in thymus and more lymphocyte arranged dose at there. Compared with the contrast group at 21,There was clear borderline between red pulp and white pulp.More germinal center and larger area of lymphocyte theca were be found in spleen;There is clear borderline between cortex and medulla and mucosa is smooth in bursa of Fabricius;The major cell was small lymphocyte in thymus and thymic corpuscle was discovered at there. Compared with the contrast group at 28,The borderline between red pulp and white pulp was clear comparatively in spleen and a lot of lymphocyte theca was formed round germinal center; Comparatively smooth mucosa and dense lymphocyte were observed in bursa of Fabricius;The major cell is small lymphocyte in thymus and the number of thymic corpuscle was became more. Be observed by electron microscope,compared with contrast group the change of ultrastructure is that there are more nucleolus of lymphocyte in trial of spleen.The unclear clromatin move to border of cell.CONCLUSION The optimal extraction process of Prinsepia Utilis Royle Flavonoids was build by orhtogonal test plan.The best extraction process was like this:ethanol concentration 60%,the ration of solid to liquid 1:20 and extracting temperature 80℃for extracting time 3h.It was identified by color reaction and the method that the mensuration of content of Prinsepia Utilis Royle Flavonoids was build up.The result showed that the method was simpie and rapid and accurate which has been applied successfully to qualitative analysis and quantitative analysis of in Prinsepia Utilis Royle flavonoidsThe test result of immunoregulation of Prinsepia Utilis Royle flavonoids in chicken showed that:Prinsepia Utilis Royle flavonoids advanced immunity of body by heighten the level blood serum antibody and it haven't bad affection on blood biochemical indicator.In addition,it advanced growth of immune organ and increased the quantity of T lymphocytes and macrophage and be beneficial to the early growth of bursa of fabricius and spleen and thymus.In conclusion, Prinsepia Utilis Royle flavonoids had advanced effect for immune response of body.