人参原型成分和代谢产物累积效应研究，以人参皂苷Rb1免疫调节作用为例

Bioavailability is an important indicator of the effectiveness of drug evaluation. Ginseng is a widely used Chinese herbal medicine with significant pharmacological activity. Its main pharmacodynamic substance are ginsenosides, however, it has proved that the bioavailability of these ginsenosides is very low which affected the objective evaluation of its effectiveness and its depth exploitation. In present study, multi-dimensional chromatography and its hyphenated mass spectrometry techniques will be used to systematically characterize the composition and content of ginsenoside Rb1 and its metabolites. In vitro biological conversion technologies will be used to investigate the metabolism of ginsenoside Rb1 such as intestinal bacteria incubation and liver microsomal incubation. Immunomodulatory activity about one-component (prototype component, a single metabolite) and multi-component (prototype and all metabolites) will be compared by using models of immunosuppressive mice and spleen lymphocytes. Therefore, the scientific hypothesis about ginseng possesses activities based on "prototype component/ active metabolites can reach the effective blood concentration through cumulative effect" will be verified. As a consequence, this project will clarify the scientific connotation of ingredients such as ginsenosides how could they possess significant pharmacological activities with the low bioavailability. By revealing the cumulative effect combined prototype component with its metabolites, this project will contribute to objective evaluation of the effectiveness and the depth development of ginseng, and also provide new ideas to explore the material basis and related mechanisms for traditional Chinese medicine.

生物利用度是评价药物有效性的重要指标之一。中药人参具有多种显著药理活性，其主要药效物质皂苷的生物利用度却很低，影响了对其有效性的客观评价和深度开发利用。本项目拟以人参皂苷Rb1为例，整合UPLC-QTOF-MS/MS 和HPLC-TQ-MS/MS等液质联用技术，全面表征血中人参皂苷Rb1及其代谢物的结构和含量；采用肠道菌温孵、肝微粒体温孵等体外生物转化技术及化学修饰法，制备获得人参皂苷Rb1的相关代谢物；选用免疫抑制小鼠模型和脾淋巴细胞模型，比较单成分（原型成分、单个代谢物）和多成分（原型及所有代谢物组合物）的免疫调节活性，用以验证如下科学假说：人参是基于“原型成分/代谢产物的累积效应达有效血药浓度”而发挥药效。通过该假说的验证，阐明人参皂苷生物利用度低却有显著药理活性的科学内涵，为人参有效性的客观评价和深度开发利用提供科学证据，同时也为中药药效物质和作用机制的研究提供新思路。

生物利用度是评价药物有效性的重要指标之一。中药人参具有多种显著药理活性，其主要药效物质皂苷的生物利用度却很低，影响了对其有效性的客观评价和深度开发利用。该项目的研究策略首先是采取“代谢物全面鉴定-多成分药代动力学-PK-PD关联性分析”三步过渡的方式，利用UPLC-QTOF-MS/MS联用分析、对照品比对等方法，全面鉴定人参皂苷Rb1入血的代谢物；建立高灵敏度、多反应监测的HPLC-TQ-MS/MS检测技术，检测在正常及病理状态下多成分的药代动力学过程；以免疫抑制小鼠物模型，建立PK-PD量效关联，阐明药物浓度-效应-时间三维关系，从而为接下来的假说验证确定人参皂苷Rb1及其代谢物的含量；然后通过对小鼠巨噬RAW264.7细胞的增殖、迁移、侵袭及炎症因子表达等指标，比较了单成分（人参皂苷Rb1原型或单个代谢物）和多成分（人参皂苷Rb1原型及所有代谢物组合物）的免疫调节活性，从而成功验证如下科学假说：人参皂苷是基于“原型成分和代谢产物的累积效应达到有效血药浓度”而发挥药效。通过该假说的验证，阐明人参皂苷生物利用度低却有显著药理活性的科学内涵，为人参有效性的客观评价和深度开发利用提供科学证据，同时也为中药药效物质和作用机制的研究提供新思路。