基于不同君药——红芪替换经典复方中黄芪抗免疫衰老机制比较研究

The aging of the population has become a serious social,medical problem,and in aging mechanism,immunosensscence is the important reason of accelerated aging,among them and T lymphocyte's structure and function changes related,through the immune adjustment can delay the ageing.Traditional Chinese medicine Radix Hedyseri and Radix Astragali all have immune adjustment function, but about difference research on anti-immunosensscence effect is less. In the course of the study show that immune adjustment exist differences about Radix Hedyseri or Radix Astragali gentleman medicine for the same compound, Radix Hedyseri compound to T cell adjustment effect was more apparent. Therefore, this research is designed through in vitro and in vivo experiments, from cell and molecular and gene angle to observe and compare the effect of Radix Astragali or Radix Astragali compoud on the thymus and spleen lymphocyte number and function, immune synapse, apoptosis and CD28 signal transduction pathway etc. It will reveal immune regulation otherness of Radix Hedyseri or Radix Astragali compound, further clarify anti-aging immune adjustment mechanism of Radix Hedyseri compound. It will also help to find out new drugs compatibility and therapeutic targets for TCM compound anti-immunosenescence, and make up for the deficiency of the theory of Radix Hedyseri pharmacological research and medical theory. On the other hand, it will settle the problem of inadequate of Radix Astragali medicinal herbs resource, and provide theory support for the comprehensive exploitation of Radix Hedyseri's medicinal value and application to spread.

人口老龄化已成为严重的社会、医学问题，而免疫衰老是加速衰老的重要原因之一，其中与T细胞的结构、功能等变化相关，通过免疫调节可延缓衰老。中药红芪与黄芪均有免疫调节作用，但两者抗免疫衰老的差异未见报道。本课题组既往研究表明用红芪与用黄芪玉屏风散含药血清均有一定的抗衰老作用，且用红芪作用较优于用黄芪玉屏风散。故本项目拟进一步通过体外和体内实验，从细胞、分子和基因角度，采用PKH26标记结合活体荧光成像、Western blot、流式细胞术、免疫组化、Southern blot、DNA双脱氧链终止法测序、荧光实时定量-PCR等技术比较研究用红芪与用黄芪复方对SAMP8小鼠胸腺细胞、微环境和TCR基因以及脾T细胞数量、功能、迁移-归巢、免疫突触及CD28信号通路的影响,揭示用红芪与用黄芪复方延缓衰老机制的差异性，为中药复方拮抗免疫衰老寻找新的药对配伍和治疗靶点，同时弥补红芪药理研究和药用理论的不足。

人口老龄化已成为严重的社会、医学问题，而免疫衰老是加速衰老的重要原因之一，其中与T细胞的结构、功能等变化相关，通过免疫调节可延缓衰老。中药红芪与黄芪均有免疫调节作用，但两者抗免疫衰老的差异未见报道。本研究以相同剂量的红芪替换黄芪，通过体内和体外实验，观察君药为红芪或黄芪的复芪止汗、玉屏风散、补中益气和益气养血方剂及其含药血清对SAMP8小鼠脾脏淋巴细胞的结构、功能、迁移-归巢、表面分子及CD28信号通路的影响。研究结果显示：①在体外实验中含红芪或黄芪的复芪止汗、玉屏风散、补中益气和益气养血方剂含药血清均能促进SAMP8小鼠脾淋巴细胞的体外增殖及IL-2等细胞因子的分泌；减少SAMP8小鼠脾淋巴细胞体外培养后衰老细胞的数量，增加淋巴细胞中CD28、p38MAPK、PI3K mRNA的表达及蛋白的活化；含红、黄芪益气养血和复芪止汗方剂能增加CD28+ T细胞的比例。②在体内实验中含红芪或黄芪的复芪止汗、玉屏风散、补中益气和益气养血方剂均能改善SAMP8小鼠脾脏T淋巴细胞亚群分化比例，增加小鼠血清中IL-2等细胞因子的含量，增加脾脏淋巴细胞中CD28、p38MAPK、PI3K mRNA及CD28、p38MAPK蛋白的表达，改善SAMP8小鼠脾脏结构；含红、黄芪补中益气汤能均对SAMP8小鼠T淋巴细胞的迁移-归巢有一定影响。③含红芪玉屏风散在促进淋巴细胞体外增殖，升高血清IL-2和淋巴细胞中CD28、p38MAPK、PI3K mRNA表达方面作用优于含黄芪玉屏风散；含红芪补中益气汤在促进IL-2分泌及淋巴细胞中CD28、p38MAPK、PI3K mRNA表达方面作用优于含黄芪补中益气汤；含红芪益气养血方剂在促进淋巴细胞体外增殖， IL-2分泌，及淋巴细胞中CD28、p38MAPK、PI3K mRNA表达方面作用优于含黄芪益气养血方剂；含红芪复芪止汗方剂在促进SAMP8小鼠脾淋巴细胞分泌IFN-γ，升高淋巴细胞中CD28、p38MAPK、PI3K mRNA表达方面作用优于含黄芪复芪止汗方剂。比较说明了额用红芪与用黄芪复方延缓衰老的差异性，弥补红芪药理研究和药用理论的不足。