基于“热者益热”的姜制影响附子药性作用机制研究

‘Treating the drug being hot nature with hot-natured assistant to strength the hot nature of the drug ’ is one of the theories of processing theories on drugs’ nature, belonging to the theory of ‘strength the nature of drugs by processing with the same nature assistant’, meaning to strength the effects of drugs by such processing methods. Processing with ginger juice is the very important path to ‘Treating the drug being hot nature with hot-natured assistant’, and processing aconitum with ginger juice is the typical piece record in the literature. But because of the unclear of mechanism of processing ,the difference and uncertainty between the application of zingiberis and ginger, and the complication of processing method, the piece of aconitum with ginger juice is facing the dangerous of disappear. The purpose of this study is to do the research on the components relating to hot nature of aconitum, such as full components, alkaloid, strong heart components and microelement, the pharmacological efficacy indicators relating to hot nature of aconitum, such as rescuing from collapse by restoring Yang, tonifying fire and helping yang, eliminating cold to stop pain, and nature effect evaluation, such as the animal behavior, micromorphology, matter energy and metabolomics. After the research, we will analyze the relationship between the components, the pharmacological effects and the hot nature effect to find the influence of the zingiberis and ginger on aconitum, to clarifying the scientific connotation of processing of aconitum with ginger juice. Based on the above researches, the processing method of processing would be optimized to a better using in TCM clinic. The results of the study will be in favor of the inheriting and development of traditional processing technology and provide the foundation of establishment of characterization of system and clarification of mechanism of processing influence on hot and cold nature of drugs.

“热者益热”是炮制药性理论重要内容之一,属于“从制”范畴，即以药性相同辅料炮制药物以协同增效。姜制是使药物“热者益热”的重要途径，性味温热的姜协同热性药物起到增效作用，姜制附子是典型代表品种，在文献中有明确记载。但由于“益热”机制不明确、干姜和生姜辅料应用的差异和不确定性、以及炮制工艺复杂性等，姜制附子濒临失传危险。课题拟在“热者益热”理论指导下，研究姜制附子热性关联物质基础（全成分、生物碱类、强心成分、多糖类和微量元素）、药效指标（回阳救逆、补火助阳、散寒止痛的药理指标）以及效应评价（动物行为、微观形态、物质能量以及代谢物组学），通过“物-效-性”的关联分析，结合干姜、生姜不同的热性影响，阐明姜制附子本质内涵，并在此基础上，优化姜制附子工艺，为其临床有效应用提供支持。研究将利于传统炮制工艺的传承与发扬，并为炮制影响中药寒热属性表征体系的构建及系统阐明炮制影响药物寒热药性科学内涵奠定基础。

研究探究了姜制附子前后全成分、生物碱、强心成分等的变化，对姜制附子进行安全性评价，并对姜制前后附子回阳救逆，补火助阳，散寒止痛三方面药效指标做比较。对成分、药性和药效关联分析，初步阐明姜制附子炮制机理。研究为姜制附子的临床应用奠定科学基础。.姜制对附子成分影响较大，但多表现为量变：姜制附子总生物碱和双酯型生物碱都有明显降低：生姜片拌蒸附子和干姜片拌蒸附子总生物碱含量下降最多，可能与米泔水漂制环节有关；姜促进了毒性成分新乌头碱水解为苯甲酰新乌头碱。5种姜制附子多糖含量不同，其中生姜榨汁制附子多糖含量最高。强心类成分去甲猪毛菜碱和尿嘧啶含量是生姜榨汁制附子最高，去甲乌药碱含量是生姜煮制附子最高。指纹图谱研究发现，与生附子相比，姜附子在总提取物的成分只有含量不同，成分的种类基本相同。.姜制可降低附子毒性，增强热性，镇痛和强心以生姜榨汁制附子效果最好：各姜制附子毒性均较生附子低；各姜制附子组大鼠舌乳头均有角质层明显增厚的现象，Na+-K+-ATP酶活性活性也都明显高于生品组，表明姜制后附子热性增强。与模型组比，生姜榨汁制附子和干姜煮汁制附子有显著镇痛作用，前者作用有极显著性差异；生附子组和生姜片拌蒸制附子抗炎作用与阳性药相近。不同姜制附子与模型组比,均可提升强心作用，但途径和强弱不同，综合表现生姜榨汁制附子强心作用最强。附子经姜制后对β肾上腺素能受体mRNA的表达减弱，并抑制心脏负性肌力作用，表明姜制附子在减弱心脏毒性的同时也保留较好强心作用。.初步阐明基于热者益热的姜制附子炮制机制：建立姜附子炮制前后药性、药效、成分关联分析法，阐明姜制附子后，改变次乌头原碱、苯甲酰乌头碱、乌头碱、脱氧乌头碱等成分含量和各成分比例，达到改变附子抗炎镇痛作用和Na+-K+-ATP酶活力值作用；炮制对附子松果灵、附子灵、尼奥灵含量的影响，对其调节血清EPI、NA、PGE2、Ang-Ⅰ值起重要作用；多根乌头碱的变化则影响了热性指标等。