基于“肝主疏泄 — 脑肠轴”探讨药对枳壳厚朴的双靶点作用及机制

From the perspective of brain gut axis - one of the biological mechanisam of "liver controlling dispersion", we will further explore the double targets effects and molecular mechanism of anti-depressive and prokinetic effects of drug pair Fructus aurantii and Magnolia bark decoction on the basis of previous researching work. Content: on the purpose of determining the optimum dose of FM for its anti-depressive and prokinetic effects, we will build chronic stress-induced depression rats model and use depression behavior and gastrointestinal motility as observation indexes to observe the anti-depressive and prokinetic effects of FM on rats model, and then, the compounds which can be absorbed by serum, the limbic system and gastrointestinal tract tissue will be determined by UPLC/MS/MS.We will discuss the influence of FM (the optimum dose) and the absorbed active components on the content of 5-HT and NPY in the serum of rats model, and also study the influence of FM (the optimum dose) and the absorbed compounds on protein expression and activity of 5-HT、5-HT1AR、5-HT2AR mRNA and NPY、NPY-Y1 receptor、NPY-Y2 receptor mRNA in the limbic system and gastrointestinal tract tissue. Significance: By studying the double targets effects and mechanism of FM for its anti-depressive and prokinetic effects, it can help interpreting the biologic mechanisms of"liver controlling dispersion" and also provide new thought and method for the pathogenesis study of depression and antidepressant drugs research.

在既往研究工作基础上，本研究从肝主疏泄的生物学基础之一"脑肠轴"角度进一步探讨药对枳壳厚朴汤剂（FM）抗抑郁促胃肠动力的双靶点作用及分子机制。内容：采用抑郁行为学指标与胃肠动力学指标相结合的方法，观察FM对慢性应激抑郁大鼠抗抑郁促胃肠动力作用，明确其最佳剂量；运用UPLC/MS/MS测定FM被抑郁大鼠血清、边缘系统及胃肠组织吸收的主要成分；通过ELISA法观察FM（最佳剂量）及主要吸收成分对脑肠轴5-HT及NPY在血清中含量的影响；运用免疫组织化学、分子生物学技术和酶谱法观察FM（最佳剂量）及主要吸收成分对5-HT、5-HT1AR、5-HT2ARmRNA及NPY、NPY-Y1受体、NPY-Y2受体mRNA在大脑边缘系统及胃肠组织蛋白表达及活性的影响。意义：通过研究FM抗抑郁促胃肠动力的双靶点作用及机制，有利于诠释中医肝主疏的生物学基础同时为抑郁症的发病机制及抗抑郁的药物研究提供新的思路。

在既往研究工作基础上，本研究从肝主疏泄的生物学基础之一“脑肠轴”角度进一步探讨药对枳壳厚朴汤剂（FM）抗抑郁促胃肠动力的双靶点作用及分子机制。内容：采用抑郁行为学指标与胃肠动力学指标相结合的实验方法，实验表明FM10g/kg及20g/kg具有较好的抗抑郁及促胃肠动力作用，FM20g/kg组优于10g/kg组，FM3g/kg组及FM30g/kg组均无效；运用 UPLC/MS/MS 测定了 FM 的主要7种成分：柚皮芸香苷、柚皮苷、橙皮苷、新橙皮苷、水合橙皮内酯、和厚朴酚及厚朴酚，同时研究表明这7种成分均可被慢性应激抑郁大鼠胃肠组织吸收，仅橙皮苷及水合橙皮内酯可被慢性应激抑郁大鼠血清及脑组织吸收。 FM（10g/kg及20g/kg）、橙皮苷（50mg/kg）及水合橙皮内酯（15mg/kg）能显著增加慢性应激抑郁大鼠血清、额叶皮层、海马区的5-HT及NPY含量。FM可上调海马5-HT1AR水平、对海马5-HT2AR没有明显作用，可上调肠平滑肌上的5-HT2AR水平，直接调控平滑肌的运动，能促进肠平滑肌收缩，对肠动力具有兴奋性作用；同时研究表明FM可上调海马及胃肠组织NPY-Y1R水平，对NPY-Y2R无影响。此研究证明了枳壳厚朴药对具有抗抑郁促胃肠动力的双重作用，同时也初步阐明了其抗抑郁促胃肠动力的主要成分及可能机制，有利于诠释中医肝主疏的生物学基础同时为抑郁症的发病机制及抗抑郁的药物研究提供新的思路。