Screening Of Immune Function Related Gene MHC B-LβⅡ And A Certain Purity Of Traditional Chinese Medicine Compound Polysaccharides On Immune Regulation In Chicken

Objective: In this study, the use of pure Chinese herbal compound polysaccharide, role in immune function related MHC B-L beta Ⅱ different gene type of chicken. Through the detection of different genotypes of chicken at different days of age the immune indexes to explore a certain purity compound polysaccharide of different genotypes of chicken immune regulating function determined on different genotypes of chicken, immune regulation effect of the optimum compound Chinese medicine polysaccharide concentration, for Chinese compound polysaccharide research in improving the immunity of chickens provide theoretical and experimental basis.Methods: 1 、 Water extraction and alcohol precipitation method, composed of angelica,Codonopsis, Radix Rehmanniae Preparata, Rhizoma Chuanxiong, hawthorn, Radix Polygoni Multiflori, 11 kinds of traditional Chinese medicine traditional Chinese medicine compound for the extraction of crude polysaccharide2、Using Sevage method after removal of the protein content of the crude polysaccharide.The polysaccharide by AB-8 macroporous adsorption resin adsorption and desorption, removal of pigment and small molecular weight impurity, coarse polysaccharide purification; by the phenol sulfuric acid method determination of purified polysaccharide content and purity of Chinese compound polysaccharide.3、Through the DNA extraction technology, PCR-SSCP technique and gene sequencing, on500 jute feather broiler chicks fed with genotype.4、Will a certain purity of Chinese compound polysaccharide, preparation of high dose(50mg / ml), middle dose(25 mg / ml), low dose(12.5 mg / ml) with three different concentrations;chicken of different genotypes, respectively, were vaccinated ND, grouping, medication,acquisition 21,42,56,70 four days of age of chicken peripheral blood, serum separation.5、ELISA method was used to detect the content of serum cytokines(IL-2, IL-4, IL-6,IFN-α,IFN- γ), the content of Newcastle disease antibody.6、ELISA method was used to detect the content of CD4, CD8 lymphocyte subsets, Ig G, Ig M content.7、The lymphocyte transformation rate was detected by MTT method, and the antibody titers of sheep red blood cells were detected by the method of coagulation and coagulation.Results: 1 、After extraction and purification, the traditional Chinese medicine compound polysaccharide with the content of 77.10% was obtained.2、A total of AA, AB, BB, AC, AD, five different genotypes.3、In the high dose group, AA genotype of the immune function indexes were higher than those of the other four genotypes; in middle dose group, genotype AB the immune function indexes were higher than other four genotypes; in the low dose group, genotype ab of the immune function related indicators were higher than other four genotypes.Conclusions: Analysis results show that the polymorphism of MHC B-L beta Ⅱ gene screening ofa compound traditional Chinese medicine polysaccharide optimum immune dose for enhancement of has a certain degree of influence, the optimal dose of different genotype chicken Chinese herbal compound polysaccharide different. At the same time, in three different concentrations of polysaccharide of Chinese drug, dose concentration to promote the best effect.