一定纯度中药复方多糖对不同MHC B-LβII 基因型鸡免疫调节作用及其作用机制研究

It is an urgent issue to be resolved of reseraching safe and effective enhancer. The typical function and the most prominent of chinese herbal medical polysaccharide (CHMP) is the role of immune enhancement and the advantages of little side effect. Now, the study of CHMP function is mainly focus on single CHMP, but not compound CHMP (cCHMP) which is rarely reported at home and abroad. Meanwhile, Polysaccharides can be processed and presented by MHC class II molecules, MHC class II gene polymorphism might affect the screening of CHMP optimum immune dose. In this project, we used a certain purity cCHMP, which has been shown to have significant immune-enhancing effect on preliminary studies, and applicated PCR-SSCP technique, cell culture, ELISA, RT-PCR, MTT and other research methods to study the relationship between MHC B-LβII gene polymorphism and the immunity of chicken, effects and mechanism of certain purity cCHMP on immunomodulatory in different MHC B-LβII genotype chickens. All of that will help to determine the optimal dosage of certain purity cCHMP in different MHC B-LβII genotype chickens, specific the pattern-recognition receptor of certain purity cCHMP on immunoregulation, elucidate its cellular signal pathway on immunomodulatory. Thus will provide theoretical ecidence for developing certain purity cCHMP as immunostimulants.

研究安全有效的免疫增强剂是函待解决的问题。中药多糖最突出的作用和优点是免疫增强作用和毒副作用小。中药多糖免疫活性研究主要集中在单味中药多糖，对中药复方多糖系统研究国内外很少报道。此外，多糖能被MHC classⅡ分子处理和呈递，MHC classⅡ基因多态性可能影响中药多糖免疫调节剂量的筛选。因此，本项目采用前期研究得到的免疫增强效果显著的一定纯度中药复方多糖，应用PCR-SSCP、细胞培养、ELISA、RT-PCR、MTT等研究手段，研究MHC B-LβII基因多态性与鸡免疫功能关系、一定纯度中药复方多糖对不同MHC B-LβII基因型鸡免疫调节作用及其作用机制，确定一定纯度中药复方多糖免疫调节剂量筛选是否受MHC B-LβII基因多态性影响，明确一定纯度中药复方多糖免疫调节作用受体，阐明其发挥免疫调节作用的细胞信号通路。为该中药复方多糖开发成免疫增强剂提供理论依据。

研究安全有效的免疫增强剂是函待解决的问题。基因多态性可能影响中药多糖免疫调节剂量的筛选。本项目采用PCR-SSCP、ELISA等方法，研究黄麻肉鸡MHC B-LβII基因多态性与免疫功能的关系，cCHMPS对不同MHC B-LβII基因型黄麻肉鸡和京红1号蛋鸡的免疫调节作用及机制。结果表明，黄麻肉鸡与免疫功能相关的基因型是AA、AB、BB、AC、AD型；各剂量cCHMPS均能提高各基因型黄麻肉鸡ND抗体效价、SRBC抗体滴度，IgG、IgM、淋巴细胞亚群及细胞因子含量，提高各基因型京红1号蛋鸡免疫器官指数、白细胞数，IBDV、H5、H9亚型抗体效价，且增强不同基因型鸡免疫力的最佳cCHMPS剂量不同；cCHMPS可能通过与TLR2或TLR4结合，激活MYD88和TRIF信号转导途径，活化NF-κB，调控细胞因子表达和分泌，改变细胞信号分子含量来调节机体免疫能力。本研究揭示了MHC B-LβII基因多态性可能会影响cCHMPS最佳免疫调节剂量的筛选，初步阐明了cCHMPS发挥免疫调节作用的受体和其信号通路。