化学生物信息技术用于中药泽泻品质评价数模构建及其实践的研究

It has always been difficult and prevalent to find a quality evaluation model that can not only represent the poly-component and multi-target integrated effects of Chinese meteria medica, but can also objectively express the efficacy and differences. The project used Chinese regulating lipid materia medica Alismatis Rhizoma as model drug, combined modern biochemical information technology, applied both experimental methods (electrochemistry and spectroscopy) and theoretical calculations (molecular simulation), and established the interaction in vitro model with the protein (Alismatis Rhizoma's effect enzyme for regulating lipid) - ligand (Alismatis Rhizoma's effect substance for regulating lipid) based on principles of Chemical Biology, after which relations between differences at parameters before and after and effect enzyme in vivo activities before and after the experiment were analyzed, so as to demonstrate its pharmacological effects and provide precise and reliable biochemical information evaluation parameters with good quantized characters. As the biological activity index, the series of parameters was used to establish mathematical model after conducting relevance analysis with effective ingredients of Chinese materia medica. Later, the mathematical model was applied for application verification of Alismatis Rhizoma sold in the markets and other Chinese materia medica. Thus, an quality evaluation model for controlling quality of Chinese materia medica with multiple indexes, great efficiency and high credibility was established, with biological activity of Chinese materia medica at the core, and supplemented by chemical ingredients included indexes, which not only meets nature of Chinese materia medica, but is also expected to provide new thinking way and technology platform for its quality evaluation.

如何建立既反映中药多组分多靶点整体效应，又能客观准确表达其功效作用及其品质差异的评价模式是当今中药学研究的热点和难点之一。本课题以调脂中药泽泻为模型药物，基于化学生物学原理，组合现代化学生物信息技术，将实验方法（电化学法、光谱法）与理论计算（分子模拟）联用，构建蛋白（泽泻调脂效应酶）-配体（泽泻调脂效应物质）的相互作用体外模型，将得出的作用前后参数的差异与效应酶作用前后的体内活性差异进行相关性分析，确定能够确切体现药效且具良好量化特点的可靠有效的评价参数。将此系列参数作为生物活性指标，与药材有效成分含量进行关联分析，建立数学模型。并对市售泽泻药材及其他中药进行应用验证。从而构建以中药生物活性为核心内容，辅以化学成分为内在指标的多指标高效率可信度高的品质评价模型。该模型符合中药本质，为其品质评价提供新的思路和技术平台。

目前中药品质评价大多是从化学角度考虑，以其化学成分作为评价指标，却忽略了本身的药理药效作用。如何建立既反映中药多组分多靶点整体效应，又能客观准确表达其功效作用及其品质差异的评价模式是当今中药学研究的热点和难点之一。本课题以调脂中药泽泻为模型药物，研究结果如下：（1）明确不同产地泽泻药材、分离到的有效部位、有效成分单体、不同配比单体混合物调血脂作用及对脂代谢关键酶三羟基三甲基戊二酰辅酶A（HMG-CoA）还原酶、脂蛋白脂酶（LPL）、卵磷脂胆固醇酰基转移酶（LCAT）、脂酰酶A胆固醇酯酰基转移酶（ACAT）的影响。（2）基于化学生物学原理，组合现代化学生物信息技术，将实验方法（凝胶电泳法、光谱法）与理论计算（分子模拟）联用，确定蛋白（泽泻调脂效应酶）-配体（泽泻调脂效应物质）相互作用的结合模式、结合部位、结合力、结合比、结合能等，建立两者相互作用的结合模型。（3）根据效应物质与效应酶相互作用模型，结合效应物质对效应酶的体内活性影响，进行相关性分析，确认泽泻醇类化合物的舵手基团为其侧链，分析得到脂代谢关键酶相互作用的关键氨基酸残基。阐述泽泻调脂效应物质与效应酶的作用机理及作用规律，筛选得到活性高的单体化合物及最佳成分配比，探讨有效组分之间的协同作用机制，并与不用产地药材有效成分含量差异进行相关分析，分析可靠有效灵敏的评价指标。选取调血脂指标表达水平为因变量，选用不同产地泽泻调脂效应物质含量及调脂效应酶活性为自变量，应用SPSS19.0统计软件，通过偏最小二乘法（PLS）模型识别方法进行相关性分析，建立不同产地泽泻调脂药效与五种调脂效应物质以及四种调脂效应酶之间的关系方程。从而构建以中药生物活性为核心内容，辅以化学成分为内在指标的多指标高效率可信度高的品质评价模型。该模型符合中药药效评价的发展方向，能客观准确表达其功效作用及其品质差异，理论上比现行的中药指标性成分定性定量分析具有优势，为其品质评价提供新的思路和技术平台。