从“成分-代谢-生物效应”关联变化探讨建昌帮阴附片、阳附片作用差异及炮制机理

Jiangchang Faction is one of traditional medicine faction in southern China originating from Jianchang in Jiangxi province. As recorded in ‘Traditional processing technologies of Jianchang Faction ’ , Yangfupian and Yingfupian were pieces of Aconite processed with different assistants and methods. The method of processing Yangfupian was to stir-friing bleached and cut salted aconite root with sand. Yangfupian was appropriate for male, especially old men, with Yang deficiency syndrome. The method of processing Yingfupian was to steaming bleached and cut salted aconite root with ginger juice. Yingfupian was appropriate for female with Yang deficiency syndrome. Although record for a long time, there is no systematic research and effective use of Yangfupian and Yingfupian in modern time. Based on our previous study on the influences and processing mechanism of hot property of Aconite processed with different ginger and guided by the traditional theory of ‘Yangfupian and Yingfupian were appropriate for male and female respectively ’, this project is going to study the differences of components, the body's absorption and metabolism, pharmacological effects and biological effects of Yangfupian and Yingfupian with LC-MS,metabonomics,proteomics andbioinformatics analysis. To discussing the effects differences andpProcessing mechanism of Yinfupian and Yangfupian based on co-relation changes of components-metabolism-biological effects through establishing Ingredients- target network, target-path network and path-effect network. It is believed that the results of the study will be significant for saving endangered processed Chinese medicine and their extensive clinical suitability application.

建昌帮是我国南方江西古药帮，据《建昌帮中药传统炮制法》记载：阳附片、阴附片是依据男女性别差异，采用不同辅料和方法得到的附子炮制品。阳附片是附子漂后以砂炒爆，以男性使用为主，尤适宜男性老人阳衰诸证；阴附片是附子漂后与姜蒸制，多用于女性肾阳虚体弱诸证。尽管古有记载，近现代并未对阳附片和阴附片开展系统研究及有效应用。因此，课题拟在前期研究不同姜制对附子热性影响及作用机制基础上，进一步以阳附片和阴附片男女异用传统理论为指导，利用液质联用、代谢组学、蛋白组学、生物信息学分析等，研究阴附片和阳附片全成分和机体吸收代谢差异、药效差异，以及由此引起的生物效应物质基础差异等，通过构建成分-靶点网络，靶点-通路网络和通路-效应网络，从成分-体内代谢-生物效应基础关联变化角度，系统揭示阳附片和阴附片男女异用的作用机制及其炮制内涵。研究对挽救濒临失传的传统特色炮制品种、促进其更广泛的临床适宜性应用具有重要意义。

研究在建昌帮附子炮制品阴附片和阳附片男女异用的传统理论指导下，开展了阴附片、阳附片工艺优化、全成分质变量变、治疗阳虚证和慢性心衰药效及其机制研究，基于棕色脂肪组织热性差异研究等。采用Box-Behnken响应面法优选工艺，明确阴附片为：生姜用量16.65%，闷润时间12h，蒸制时间9.5h，阳附片为：砂量 20%，炒制温度150℃，时间5min。去皮能有效去除盐附子盐分，并较传统漂法显著保留有效成分。采用 UPLC-Q/TOF-MS对成分变化进行分析，确定了Chuanfumine、Senbusine A等27个差异性成分。生附子双酯型生物碱占比较高，阴附片胺醇型生物碱占比较高，阳附片单酯型生物碱占比较高，提示阴附片蒸制更有利于酯键的水解，阳附片爆炒使双酯型生物碱快速大量降解为单酯型。性别差异药效研究表明，阴附片和阳附片对动物性腺组织、性激素、免疫水平以及环核苷酸系统有较大性别作用差异，阴附片对阳虚雌性大鼠卵巢和子宫形态有明显改善，阳附片对阳虚雄性大鼠曲细精管形态有明显改善，在血清促卵泡素、血清雌二醇含量，免疫球蛋白水平及性组织形态等方面，也均存在较大差异。代谢组学研究发现，阴附片对阳虚证主要作用为上调线粒体核糖体蛋白等，而阳附片和生附子为下调（辅）肌动蛋白，提示阴附片、阳附片男、女异用理论的科学性。强心作用表明，两者能有效改善心衰大鼠心肌组织细胞空泡、组织间隙增宽断裂等情况。通过下丘脑 AMPK 蛋白-棕色脂肪组织-脂质代谢信号通路研究，验证了阴附片和阳附片对棕色脂肪组织不同的热性影响。研究验证了建昌帮附子炮制品阴附片和阳附片男女异用的传统炮制理论，揭示其科学内涵。发表论文9篇，获省级科技进步一等奖2项，专利1项，获批省级双千计划1人，主持承担了江西省建昌帮特色技术炮制规范研究等揭榜挂帅项目2项。研究为传统特色炮制品种的现代研究提供了可行思路，为特色饮片品种转化应用奠定坚实基础。