盐巴戟补肾壮阳作用增强的药效物质基础及炮制机理的研究

Under the theory "Tonic decoction should use prepared-medicine to get the best effect"and "salt-prepared could lead medicine into the renal", What is the material base for Morinda officinalis steamed by salt？ why the effect of strengthening the kidney and invigorating the yang is better for Morinda officinalis after being steamed by salt？ We apply "processing-material base-pharmacology, pharmacodynamics-pharmacokinetics-absorption" combined with metabonomics as research model. Firstly, we separate and isolate chemical components by chromatographic methods and identify their structure by spectral methods from iridoids glycosides and oligosaccharides which exhibit considerable variation after steaming with salt, thus explain the processing principle from the perspective of material base. Secondly, with the theory of the neuro-endocrine-immuno-modulation network, we observe how Morinda officinalis steamed by salt regulate the function of hypothalamic-pituitary-adrenal axis, hypothalamic-pituitary-thyroid axis, hypo- -thalamus-pituitary-gonad axis and immune regulation by some pharmacodynamics experiments. Thus illustrate why the effect of invigorate the kidney and strengthen yang is more obvious after being steamed by salt. Thirdly, to explain the kidney-trend of Morinda officinalis after being steamed by salt, we conducted pharmacokinetics and tissue distribution experiments for active components of Morinda officinalis steamed by salt to illustrate Morinda officinalis after being steamed by salt could lead the drug into the kidney. Fourthly, we investigate the absorption characteristic of active components from Morinda officinalis steamed by salt to denote whether Morinda officinalis steamed by salt could enhance the intestinal absorption, that is to say, active components of Morinda officinalis steamed by salt had better biological effects. Finally, combined urine and blood plasma metabonomics experiments by 1H-NMR, we can explain the processing principle for Morinda officinalis steamed by salt comprehensively.

课题在"补汤宜用熟""入盐走肾脏"理论指导下，研究盐巴戟补肾壮阳的物质基础及药效变化机制。采用"炮制-物质-理效-药动-吸收" 结合代谢组学的研究模式。采用柱层析、制备液相、制备薄层等手段，对巴戟天盐炙后变化较大的环烯醚萜苷类、寡糖类成分进行系统分离，光谱法鉴定化合物结构，从物质基础方面说明巴戟天盐炙炮制机理；以神经内分泌免疫网络学说为指导，考察盐巴戟对肾阳虚动物的下丘脑-垂体-肾上腺轴、下丘脑-垂体-甲状腺轴、下丘脑-垂体-性腺轴调节、分子免疫调节等功能的改善，以此说明盐巴戟补肾壮阳作用增强机制，解释"补汤宜用熟-盐巴戟补肾壮阳"理论；通过体内代谢动力学、组织分布等实验，说明盐巴戟作用趋向的变化，解释"入盐走肾脏-盐巴戟入肾经"理论；通过肠吸收实验说明盐巴戟的活性成分小肠吸收机制，考察盐巴戟是否能促进活性成分的吸收；最后结合尿液、血液代谢组学实验，全面解析盐巴戟的炮制机理。

本课题在“补汤宜用熟”，“入盐走肾脏”的传统理论指导下，对盐巴戟补肾壮阳作用增强的物质基础及药效机制进行了研究。采用炮制-物质-理效-药动-吸收-代谢组学的研究模式，以炮制化学-药效机制-入血成分和代谢组学的系列完整的研究，解析了盐巴戟补肾壮阳作用增强的作用机制。以UPLC-Q-TOF对巴戟天盐蒸前后的成分进行定性分析，共鉴定出116个化学成分，采用OPLS-DA及S-plot筛选出22种潜在化学标记物；对蒽醌和环烯醚萜苷类成分进行了化学成分系统分离，鉴定出10种化学成分，并对茜草素型蒽醌和环烯醚萜苷类成分的质谱裂解规律进行了研究；采用UPLC-QqQ-MS对巴戟天中的10种化学成分进行了含量测定；对盐蒸过程中环烯醚萜苷类成分的变化规律进行了研究。采用正交设计和星点效应面法对巴戟天的软化切制工艺，盐蒸工艺进行了考察，优选了最佳的工艺参数。以免疫神经内分泌网络学说为指导，采用ELISA方法、PCR、免疫组化等手段对下丘脑-垂体-靶腺轴（肾上腺轴、甲状腺轴、性腺轴），免疫调节、改善能量代谢、改善生殖功能等方面的激素水平变化做为指标进行检测分析，全面说明盐巴戟补肾壮阳作用增强的作用机制。通过体内药动学实验以及分子药理学实验对巴戟肉、盐巴戟的有效成分甲基异茜草素、甲基异茜草素-1-甲醚、水晶兰苷的组织分布和药代动力学进行了研究；建立Caco-2细胞模型以及大鼠在体肠灌流模型实验，从细胞、在体两个层次研究甲基异茜草素-1-甲醚、耐斯糖、水晶兰苷等主要活性成分的肠吸收代谢机制。同时采用uplc-Q-TOF技术对巴戟天盐蒸前后的入血成分和代谢物进行了考察。对巴戟天盐蒸前后对肾阳虚大鼠的血清和尿液进行了代谢组学分析，采用PCA和OPLA法比较潜在生物标志物的含量差异，找到变化较大的代谢差异物及代谢通路，研究巴戟天不同炮制品对大鼠肾阳虚症的干预效果，解析巴戟天补肾阳作用的机制。