女贞子酒蒸增效机理研究

Steamed Ligustri Lucidi Fructus can nourish liver and kidney, but the processing mechanism has not been clarified. Our previous study has drawn two conclusions. The first, the contents of specnuezhenide and oleuropein reduced after steamed. Conversely, the contents of salidroside, nuezhenidic acid and hydroxytyrosol increased significantly. The Second, the absorptivity of specnuezhenide was much lower than salidroside in the stomach and the small intestine of rats. Specnuezhenide can be metabolized to salidroside in the gastrointestinal tract of rats. Based on the above conclusions we can infer that parts of the Secoiridoid Glucosides in Ligustri Lucidi Fructus can be hydrolyzed into its secondary glycosides or aglycones. The hydrolysis products are more beneficial to be absorbed. Therefore, the efficacy of Ligustri Lucidi Fructus can be enhanced. This project intends to use HPLC fingerprint comparison, macroporous resin, reverse phase silica gel column chromatography and high performance liquid preparative chromatography to separate and prepare characteristic components which are changed after steamed in terms of both quality and quantity. We will identify the structure by IR, MS, NMR, HMBC and so on to. In the wine steaming process we can use HPLC,UPLC-Q-TOF-MS and analog processing technology of monomer component to study dynamic change and transformation mechanisms of the components and to explore the absorption and metabolism of rat in vivo differences. All the above can clarify synergistic mechanism of steamed Ligustri Lucidi Fructus from the perspectives of active materials conversion, absorption and metabolism which can provide scientific basis for the processing procedures, quantitative standard for Ligustri Lucidi Fructus.

酒蒸女贞子可增强其滋补肝肾作用，但其炮制机理尚未阐明。我们前期研究发现，女贞子酒蒸后，特女贞苷、橄榄苦苷含量降低，而红景天苷、女贞酸、羟基酪醇含量显著升高；特女贞苷在大鼠胃与小肠的吸收率明显低于红景天苷，特女贞苷在大鼠胃肠道内均可代谢为红景天苷。据此推测"酒蒸女贞子可能使其部分环烯醚萜苷类成分水解，生成其次级苷或者苷元更利于机体吸收，而达到增强其药效的目的"。本课题拟运用HPLC指纹图谱对比，大孔树脂、反相硅胶、凝胶等柱色谱及高效液相制备色谱等方法，分离制备女贞子酒蒸前后质变与量变特征成分，经IR、MS、NMR、HMBC等技术鉴定其结构；采用HPLC、UPLC-Q-TOF-MS及单体成分模拟炮制技术，研究该类成分在酒蒸过程中的动态变化与转化机制，探讨其在大鼠体内的吸收与代谢差异；从活性成分转化与吸收代谢角度揭示女贞子酒蒸增效的机理，为制定女贞子的炮制工艺与质量标准提供科学依据。

[项目背景] 女贞子为中医临床常用的滋补肝肾中药。长期的临床实践表明，酒蒸女贞子可缓和其寒滑之性，增强滋补肝肾作用，故女贞子历代入药均以炮制品为主，但其炮制机理尚未阐明，致使其炮制工艺和质量标准缺乏专属性科学依据，严重影响了女贞子饮片的质量和临床疗效。本课题主要研究女贞子酒蒸前后质变与量变特征成分的实质、变化规律与转化机制及其在大鼠体内吸收代谢差异，从活性成分转化与吸收代谢角度探讨女贞子酒蒸增效机理。[研究内容] 女贞子不同炮制品对肾阴虚模型大鼠睾丸组织氧化应激损伤的保护作用比较；女贞子生品与酒蒸品化学成分的HPLC-ESI/MS分析；女贞子酒蒸前后质变与量变特征成分的分离与鉴定；女贞子酒蒸过程中质变与量变特征成分含量变化规律研究；女贞子不同炮制品HPLC指纹图谱色谱峰的归属与比较；女贞子酒蒸过程中质变成分转化机制研究；女贞子酒蒸前后质变与量变特征化合物的大鼠在体胃肠吸收比较；女贞子酒蒸前后质变特征化合物的大鼠胃肠代谢物检测；女贞子酒蒸前后质变与量变特征化合物的大鼠血浆药动学比较。[研究结果] 女贞子经酒蒸后对肾阴虚模型大鼠睾丸组织氧化应激损伤的保护作用增强；从生女贞子中分离出14种化合物，鉴定了其结构；女贞子酒蒸后环烯醚萜苷类成分产生质变，苯乙醇类成分产生量变；女贞子中含有酯苷键的环烯醚萜苷类成分性质不稳定，加黄酒（或加水）、加热可使其裂环上的酯键及环烯醚萜部分的氧苷键水解，生成其次级苷或者苷元；水解后的产物（苯乙醇苷或苯乙醇苷元）具有较强的生物活性，且分子量变小，较其原苷更利于机体吸收，从而达到增强其药效的目的；酒蒸水解的环烯醚萜苷类成分在大鼠体内也可代谢为其次级苷或者苷元，与炮制过程中的转化规律基本一致。[科学意义] 本课题创新性地从活性成分转化与吸收代谢角度阐明了女贞子酒蒸增效机理，丰富了中药炮制增效理论，为制定女贞子饮片的炮制工艺和专属性质量标准提供了科学依据。