柴胡疏肝散吸收成分在大脑边缘系统-肠浓度与抗抑郁促动力的关系

The colleagues in our research group, which broaden the study the National Natural Science Foundation of China "The study of molecular mechanism of Xiaoyaosan Xiaoyaosan to antidepressant and promote gastrointestinal motility based on abnormal brain-gut axis-stagnation and spleen deficiency pathway"(No.81072967), found Chaihu-Shugan-San(CSS) has similar antidepressant effect as Xiaoyao-San(XS), and promotes gastrointestinal stronger. The pilot studies confirmed that entering the hippocampus/bowel composition of CSS, ferulic acid(Chuanxiong) and merazin hydrate(Zhiqiao), played an important role. The project plans, with "blood/ limbic system /intestinal absorption prescriptions components-pharmacodynamics-target organ concentration and effect" for research ideas, to explore material base of CSS antidepressant and promoting gastrointestinal motility, and clarify relationship and mechanism of target organ efficacy composition concentration and curative effect. First, using UPLC-MS/MS to determinate multiple active components entering blood/ limbic system /intestine of depression with gastrointestinal disorders/model animals which oral administrated/irrigated stomach CSS. And then, using depression ethology, gastrointestinal dynamics which recognized as the index for the curative standard, to determine the effect material of original formula and elucidate the mechanism. Finally, comparating curative effect of formula/ active components alone or combined to administration, to determine the optimal dose and clear relations of dose-target organ concentration-best effect, to provide the experimental evidence for effective and safe clinical application of CSS.

经拓宽本课题组同事正负责研究的国家自然基金项目"从脑肠轴异常-肝郁脾虚途径研究逍遥散抗抑郁促胃肠动力的分子机制研究"（批准号：81072967），发现柴胡疏肝散与逍遥散抗抑郁疗效类似，且促胃肠动力更强。前期研究证实方中进入海马/肠成分阿魏酸（川芎）和水合橙皮内酯（枳壳）起了重要作用。本项目拟以"血/边缘系统/肠吸收方剂成分-药效学-靶器官浓度与效应"研究思路探索该方抗抑郁促动力疗效的物质基础，阐明靶器官药效成分浓度与疗效的关系及作用机制。首先以UPLC-MS/MS测定抑郁症伴胃肠动力障碍患者/模型动物口服/灌胃该方后吸收入血/边缘系统/肠的多种活性成分；进而以抑郁症行为学，胃肠动力学等公认的指标为疗效标准，确定母方效应物质并阐明其作用机理；最后经比较复方/药效成分单独或组合给药疗效确定最佳复方/药效成分剂量，明确剂量-靶器官浓度-最佳效应关系，为柴胡疏肝散有效、安全的临床应用提供实验依据。

柴胡疏肝散能明显改善大鼠的抑郁行为及胃肠功能障碍，本实验首先运用UPLC检测柴胡疏肝散汤剂中6种成分的浓度，并运用LC-MS/MS检测吸收入血/肠/脑成分，结果示阿魏酸、柚皮苷、橙皮苷、水合橙皮内酯和甘草素5种成分吸收入血和肠，阿魏酸和水合橙皮内酯吸收入脑。运用抑郁症行为学和在体胃肠动力检测研究吸收入血和肠5种成分的抗抑郁促胃肠动力作用，结果示：水合橙皮内酯和阿魏酸与柴胡疏肝散在改善大鼠抑郁及促胃肠动力方面有相同的趋势，橙皮苷，柚皮苷和甘草素能够部分改善大鼠抑郁行为及胃肠动力障碍。分子机理研究显示：柴胡疏肝散及吸收入血成分通过HPA轴途径，神经递质/肽，胃肠道激素，一方面作用于脑边缘系统抗抑郁，一方面作用于消化道改善胃肠动力，吸收入血成分橙皮苷可下调模型大鼠血浆皮质酮（CORT）水平，阿魏酸上调模型大鼠脑内单胺递质去甲肾上腺素（NA）、5-羟色胺（5-HT）和多巴胺（DA）含量，水合橙皮内酯可上调5-HT水平，阿魏酸和橙皮苷可增加模型大鼠血清MLT水平。行为学及分子机理显示：阿魏酸、柚皮苷、橙皮苷、水合橙皮内酯和甘草素为母方柴胡疏肝散抗抑郁促胃肠动力的药效成分。进一步运用UPLC检测柴胡疏肝散5种成分的血药浓度，阐明其药物动力学特征，揭示柴胡疏肝散剂量-效应成分血药浓度-药效关系，为柴胡疏肝散安全有效的临床应用提供实验数据。