基于免疫细胞因子调控的补气名方“玉屏风散”配伍机理与作用机制研究

The immune system is a system of biological structures and processes within an organism that protects against disease. To function properly, an immune system must detect a wide variety of agents, from viruses to parasitic worms, and distinguish them from the organism's own healthy tissue. The immune system works with cells, tissues and organs in a coordinated manner, which is able to attack pathogens with a complex array of defensive mechanisms, i.e. defense, surveillance and tolerance, like a network. The new generation of immune modulatory drugs should have multiple targets to strike the balance of the mediators. Thus, TCM herbal formulae, which are characterized by the merits of “multi-drug and multi-target”, are more suitable for this criterion as compared to synthetic western drugs. In TCM theory, “Qi” is the vital energy. There is no doubt that “Qi” is related to the defensive function of our body. Therefore, immunology has long been well-recognized as an important part of human physiology in TCM. Yu Ping Feng San (YPFS; Jade screen powder), a Chinese herbal decoction comprised of Astragali Radix (AR; Huangqi), Atractylodis Macrocephalae Rhizoma (AMR; Baizhu) and Saposhnikoviae Radix (SR; Fangfeng). YPFS was designed to boost “Defensive Qi” and arrest sweating by closing the skin pores. Clinically, YPFS has been shown to have beneficial immune-modulatory effects in preventing bacterial/viral infections and promising curative effects in chronic inflammation diseases. However, the standardization on YPFS is limited; the active ingredients that account for its functions are unclear and the action mechanisms of YPFS are unknown. Our previous studies have chemically standardized YPFS and proved YPFS possessed the ability to stimulate the cytokines release in cultured macrophage. Therefore, in this study, we aim to set up the quality control of YPFS, investigate the cellular effects and the underlying action mechanisms, thus finding out the active ingredient.

目前亚健康造成的免疫力低下正向中青年人群快速扩散。玉屏风散是中医临床提高免疫力的经典方剂，课题组前期研究表明，玉屏风散可有效促进巨噬细胞分泌免疫细胞因子，从而对免疫系统进行调节，但是还未阐明该方对免疫系统的作用机理和药效物质。因此，本课题拟从玉屏风散化学成分入手，对玉屏风散的体外化学分析和体内入血成分及其代谢物进行分析；采用动物和细胞模型，测定免疫器官重量、检测免疫分子如白细胞介素、干扰素和肿瘤坏死因子表达和巨噬细胞的数目，在此基础上采用分子生物学手段，对巨噬细胞系和血管内皮细胞中免疫分子表达进行研究，推测玉屏风散在巨噬细胞系和血管内皮细胞中免疫分子表达的信号通路。将化学成分与药理实验进行相关分析，探讨玉屏风散的配伍意义，并从“整体动物-细胞-分子”水平全面阐明玉屏风散调控免疫系统中免疫细胞因子的药效物质与作用机理，为开发提高免疫力药物与揭示复方中药药效物质与作用机理提供科学依据。

玉屏风散是中药益气固表止汗的经典补气代表方剂，在现代医学中广泛用于临床提高免疫力。本研究采用现代科学技术手段从化学和生物两个方面展开，建立了中药复方玉屏风散的化学标准并全面的分析了玉屏风散提高免疫功能的分子机制及其在对抗癌症耐药性方面的功能，研究成果如下：（1）选取玉屏风散中15种代表性成分为指标化合物，利用RRLC-QQQ-MS/MS，建立玉屏风散提取物的指纹图谱，同时利用HPLC-DAD-QQQ-MS/MS，建立了质量控制标准；（2）发现玉屏风散可以调控白介素等免疫因子的表达和释放并促进巨噬细胞吞噬细菌的功能从而提高机体的免疫功能；（3）在细胞和动物模型中证明玉屏风散可以调控多药耐药性蛋白并逆转抗癌药顺铂的肿瘤耐药性。本研究采用了先进的化学分析和生物学研究手段，从整体动物到细胞分子，多层次的阐释了中药复方玉屏风散的免疫增强和协同抗癌作用，为对抗顺铂耐药性的相关研究提供了重要依据。上述成果培养了一名博士生毕业，发表了一篇SCI论文。