硒化修饰提高中药多糖的增强免疫和抗氧化活性及其机理研究

On the basis of the previous and previous preliminary researchs which confirmed that sulfating modification could significantly enhance the antiviral and immune-enhancing activities and selenizing modification could significantly enhance the anti-oxidative and immune-enhancing activities of Chinese herbal medicinal polysaccharides, this project will carry out following researches:.Firstly, ten kinds of Chinese herbal medicinal polysaccharides will be selected as research object and the total and fractional polysaccharides of each kind of polysaccharide will be extracted respectively from the corresponding Chinese herbal medicines..Secondly, the active sites of every kind of polysaccharide will be traced taking the immune-enhancing and anti-oxidative activities as the index..Thirdly, selenizing modification methods for each kind of polysaccharide will be established and the active sites of each kind of polysaccharide will be modified, respectively..Fourthly, the activity changes of each kind of polysaccharide before and after modification will be compared systematically by means of modern technique on pharmacology, immunology, molecular biology and so on..Fifthly, the selenizing polysaccharides with strongest activity in every kind of polysaccharide will be picked out and their mechanisms of action will be claritied from molecular, cellular and clinical aspect..Finally, the total comparison will be carried out and the selenizing polysaccharide with strongest activity will be picked out..The object of this research is to discover polysaccharid active lead compound with immune-enhancing and anti-oxidative activity and to establish the basis for development and exploitation of the new type of immunopotentiators and feed additive with high efficacy, safety and independent intellectual property right..This research possesses not only the important strategical significance for exploiting and utilizing botanic resource, exploiting new pharmacal resource and ensuring the sustainable development of the traditional Chinese medicine, but also the extensive application prospect for developing new veterinary drugs, enhancing the prevent-control effect of animal loimia and promoting the development of national economy.

本项目拟在前期研究证明硫酸化修饰能显著提高中药多糖的抗病毒和增强免疫活性、前期预研证明硒化修饰能显著提高中药多糖的抗氧化和增强免疫活性的基础上，选择10种中药多糖为研究对象，分别从相应中药中提取总多糖和分级多糖；以增强免疫活性和抗氧化活性为指标，追踪活性部位；建立各多糖的硒化修饰方法，对各多糖活性部位进行硒化修饰；运用药理学、免疫学、分子生物学等现代技术，系统比较修饰前、后各多糖的活性变化；筛选出各多糖中活性最强的硒化多糖，并从分子、细胞和临床学方面阐明其作用机制；最后进行总体比较，筛选出活性最强的硒化多糖，以发现多糖类增强免疫和抗氧化活性先导化合物，为研制开发高效、安全、具有自主知识产权的新型免疫增强剂和饲料添加剂奠定基础。本研究不仅对开发利用植物资源、开辟新药源、保障中药的可持续发展具有重要的战略意义，而且对研制开发新兽药、提高动物疫病的防控效果、促进国民经济的发展具有广阔的应用前景。

完成了当归多糖（CAP）、党参多糖（CPP）、枸杞多糖（LBP）、大蒜多糖（GBP）等12种多糖研究。首先，各多糖分别提取总多糖和分级多糖，追踪增强免疫增强和抗氧化最强的活性部位；第二，对多糖硒化修饰方法进行优选，筛选出最佳的硝酸-亚硒酸钠法，建立了各多糖的修饰方法，正交设计9种修饰条件，对各多糖活性部位进行硒化修饰；第三，通过体内外试验分别比较了硒化多糖的增强免疫和抗氧化活性，筛选出各多糖中活性最强的硒化多糖及其最佳修饰条件；第四，研究了硒化多糖增强免疫和抗氧化的作用机理，证明硒化多糖通过促进IL-2、IL-6、IFN-γ等细胞因子的mRNA表达而发挥增强免疫作用，通过影响MAPK信号通路和线粒体凋亡信号通路蛋白的表达而发挥抗氧化作用；第五，对筛选出的硒化多糖通过体内外试验进行总体比较，筛选出增强免疫活性最强的5号硒化党参多糖sCCP5、抗氧化活性最强的6号硒化枸杞多糖sLBP6；最后，应用筛选出的多糖和硒化多糖进行组方和活性比较，分别筛选出增强免疫作用最强的复方GBP-sCPP、抗氧化作用最强的复方CAP-sLBP。.通过以上研究，证明硒化修饰能显著提高多糖的增强免疫和抗氧化活性，可以作为提高多糖生物活性的有效措施；筛选出的多糖和硒化多糖，可以作为增强免疫和抗氧化的活性先导化合物，筛选出的多糖-硒化多糖复方，可以作为免疫增强剂和抗氧化剂的候选方。本研究不仅对开发利用植物资源、开辟新药源、保障中药的可持续发展具有重要的战略意义，而且对研制开发新兽药、提高动物疫病的防控效果、促进国民经济的发展具有良好的应用前景。