基于“药效←配伍→ADME”研究6个不同配比开心散类方的特异性靶标及ADME特性

Kai-xin San is a famous traditional Chinese medicine used for the treatment of emotional disease, there are six formulas with the same composition of medicinal plants and different compatibility. Our previous studies have found that the six Kai Xin San prescriptions (KXSs) showed some similar pharmacologic actions, and also had characteristic target respectively (or good at anti -depression and anxiety, or target to improve learning and memory capacity),we supposed that this characteristic is closely related to the different compatibility of the six KXSs.Here, we intend to observe the effects of the six KSXs on the specificity index of Alzheimer's disease (AD) and depression-specific pharmacodynamic indicators, and the targets of signal transduction pathway by using the classic depressive animal models and AD animal models. To determine the blood drug level and pharmacodynamic indexes of their representative composition at different time points, identify the correlation between pharmacodynamic indicators (behavior, neurotransmitters, neurotrophic factors, oxidative stress, immune regulation and ERK/MAPK and CaMK signaling pathways) and indications components, to clarify the relationship between "pharmacodynamics-compatibility" of the six KXSs. Based on the LC-MS and GC-MS technique, to explore quantitative detection methods, research the absorption, tissue distribution, metabolism by CYP450 and biliary excretion characteristics of the six KXSs and their representative components, to clarify their "compatibility-ADME" relationship. Via analyzing the findings of "pharmacodynamics-compatibility" and "compatibility-ADME", to clarify the "Pharmacodynamics-compatibility-ADME" relationship of the six KXS formulas ultimately.

课题组前期研究表明，组方相同，配比不同的开心散类方的体内外药理活性有相似之处，但各有侧重（或长于抗抑郁焦虑，或长于提高学习记忆能力等），这与其组方相同而配比不同的特点密切相关。本项目拟以6个开心散类方为研究对象，在经典抑郁模型和阿尔兹海默症模型上，观察它们对抑郁症和阿尔兹海默症特异性药效指标及信号通路靶点的影响；测定它们及其代表性成分的血药浓度和各药效指标不同时间点的经时变化，明确药效指标（行为学、神经递质、神经营养因子、氧化应激、免疫调节及ERK/MAPK和CaMK信号通路）与指征成分的关联；阐明6个类方的"药效-配伍"的关系。基于LC-MS和GC-MS技术，建立定量检测方法，研究6个开心散类方及代表性成分的吸收、组织分布、肝药酶代谢及胆汁排泄特点，阐明它们的"配伍-ADME"的关系。综合"药效-配伍"和"配伍-ADME"的研究结果，最终阐明开心散类方"药效-配伍-ADME"的关系。

开心散类方由“远志、人参、茯苓、石菖蒲” 四味药材组成，其药方组成完全相同而配伍比例不同，主治的描述虽有相似之处，主要有调节情志，益智、防老抗衰等功效，其实却各有侧重。本课题研究上述6个开心散类方对抑郁和AD模型动物行为学和与病症相关的生化指标的影响，比较它们的药理作用及可能的作用机制，初步阐明其药效与配伍之间的关系。研究结果显示，《备急千金要方》之定志小丸的抗抑郁作用确切，能够明显改善模型大鼠的抑郁症状，升高模型大鼠中枢单胺类神经递质的含量，调节模型大鼠中枢兴奋性和抑制性氨基酸的异常紊乱，抑制模型大鼠HPA轴的亢进及降低血清中细胞因子的含量；《古今录验》之定志丸也显示了较好的抗抑郁作用，并且参与了抑郁症发病机制的多个环节；其他类方的抗抑郁作用在本次研究中无法确定。类方中化学成分的差异直接影响抗抑郁作用的发挥，差异性的化学成分主要来自人参和远志，类方中人参和远志的含量差异对于抗抑郁作用的影响很大。在阿尔兹海默症的细胞和动物模型上，评价了6首开心散类方的药理作用和机制，发现《千金要方》之开心散和《医门方》之开心丸对阿尔兹海默模型的药理作用优于其他开心散类方，其作用机制主要与抑制Tau蛋白过度磷酸化、升高Ach含量和降低AchE活性密切相关，而与升高BDNF含量、降低Aβ含量和血清中NT-proBNP含量相关性较弱。药材组成相同，配比不同的6首类方中，《千金要方》之开心散和《医门方》之开心丸中茯苓和石菖蒲的配比增加或减少可能是影响其发挥治疗AD的因素之一。化学成分分析结果表明，《备急千金要方》之定志小丸和《古今录验》之定志丸的化学成分相似；而《医门方》之开心丸和《千金要方》之开心散的化学成分与其他类方的化学成分差异较大。《医门方》之开心丸与《备急千金要方》之定志小丸差异较大的是人参中的6个成分和远志中的2个成分，《千金要方》之开心散与《备急千金要方》之定志小丸差异较大的是人参中的6个成分。